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Sulforaphane is considered the bioactive metabolite of glucoraphanin after dietary consumption of broccoli sprouts. Although both molecules pass through the gut lumen to the large intestine in stable form, their biological impact on the first intestinal tract is poorly described. In celiac patients, the function of the small intestine is affected by celiac disease (CD), whose severe outcomes are controlled by gluten-free dietary protocols. Nevertheless, pathological signs of inflammation and oxidative stress may persist. The aim of this study was to compare the biological activity of sulforaphane with its precursor glucoraphanin in a cellular model of gliadin-induced inflammation. Human intestinal epithelial cells (CaCo-2) were stimulated with a pro-inflammatory combination of cytokines (IFN-γ, IL-1β) and in-vitro-digested gliadin, while oxidative stress was induced by H2O2. LC-MS/MS analysis confirmed that sulforaphane from broccoli sprouts was stable after simulated gastrointestinal digestion. It inhibited the release of all chemokines selected as inflammatory read-outs, with a more potent effect against MCP-1 (IC50 = 7.81 µM). On the contrary, glucoraphanin (50 µM) was inactive. The molecules were unable to counteract the oxidative damage to DNA (γ-H2AX) and catalase levels; however, the activity of NF-κB and Nrf-2 was modulated by both molecules. The impact on epithelial permeability (TEER) was also evaluated in a Transwell® model. In the context of a pro-inflammatory combination including gliadin, TEER values were recovered by neither sulforaphane nor glucoraphanin. Conversely, in the context of co-culture with activated macrophages (THP-1), sulforaphane inhibited the release of MCP-1 (IC50 = 20.60 µM) and IL-1β (IC50 = 1.50 µM) only, but both molecules restored epithelial integrity at 50 µM. Our work suggests that glucoraphanin should not merely be considered as just an inert precursor at the small intestine level, thus suggesting a potential interest in the framework of CD. Its biological activity might imply, at least in part, molecular mechanisms different from sulforaphane. [ABSTRACT FROM AUTHOR]

Broccoli is a rich source of diverse bioactive compounds, but how their contents are influenced by different growing seasons and variations in broccoli head sizes remains elusive. To address this question, we quantified sixteen known bioactive compounds and seven minerals in broccoli with varying head sizes obtained in two different growing seasons. Our results suggest that the contents of vitamin C, total phenols, carotenoids, and glucoraphanin were significantly higher in samples from the summer–autumn season, showing increases of 157.46%, 34.74%, 51.80%, and 17.78%, respectively, compared with those from the winter–spring season. Moreover, chlorogenic acid is a phenolic compound with relatively high contents among the six detected, while beta-sitosterol is the sterol with relatively high contents. Further, principal component analysis was conducted to rank the comprehensive scores of the profiles of phenolic compounds, phytosterols, and minerals, demonstrating that the broccoli samples grown during the summer–autumn season achieved the highest composite scores. Our results indicate that broccoli heads from the summer–autumn season are richer in a combination of bioactive compounds and minerals than those from the winter–spring season based on the composite score. This study extends our understanding of the nutrition profiles in broccoli and also lays the foundation for breeding broccoli varieties with improved nutrition quality. [ABSTRACT FROM AUTHOR]

Intestinal permeabilization is central to the pathophysiology of chronic gut inflammation. This study investigated the efficacy of glucoraphanin (GR), prevalent in cruciferous vegetables, particularly broccoli, and its derivative sulforaphane (SF), in inhibiting tumor necrosis factor alpha (TNFα)-induced Caco-2 cell monolayers inflammation and permeabilization through the regulation of redox-sensitive events. TNFα binding to its receptor led to a rapid increase in oxidant production and subsequent elevation in the mRNA levels of NOX1, NOX4, and Duox2. GR and SF dose-dependently mitigated both these short- and long-term alterations in redox homeostasis. Downstream, GR and SF inhibited the activation of the redox-sensitive signaling cascades NF-κB (p65 and IKK) and MAPK ERK1/2, which contribute to inflammation and barrier permeabilization. GR (1 μM) and SF (0.5–1 μM) prevented TNFα-induced monolayer permeabilization and the associated reduction in the levels of the tight junction (TJ) proteins occludin and ZO-1. Both GR and SF also mitigated TNFα-induced increased mRNA levels of the myosin light chain kinase, which promotes TJ opening. Molecular docking suggests that although GR is mostly not absorbed, it could interact with extracellular and membrane sites in NOX1. Inhibition of NOX1 activity by GR would mitigate TNFα receptor downstream signaling and associated events. These findings support the concept that not only SF, but also GR, could exert systemic health benefits by protecting the intestinal barrier against inflammation-induced permeabilization, in part by regulating redox-sensitive pathways. GR has heretofore not been viewed as a biologically active molecule, but rather, the benign precursor of highly active SF. The consumption of GR and/or SF-rich vegetables or supplements in the diet may offer a means to mitigate the detrimental consequences of intestinal permeabilization, not only in disease states but also in conditions characterized by chronic inflammation of dietary and lifestyle origin.

Lepidium draba (hoary cress) is a perennial plant belonging to the Brassicaceae family that produces two dominant glucosinolates (GLSs): glucoraphanin (GRN) and sinalbin (SBN). They represent the stored form, which is converted upon the myrosinase (Myr) hydrolysis activity to active compounds, mainly isothiocyanates (ITCs) such as sulforaphane (SFN) or p-hydroxybenzyl isothiocyanate (pHBITC). Research on ITCs that have proven anticancer, antimicrobial, and chemoprotective properties is usually conducted with pure commercially available compounds. However, these are chemically reactive, making it difficult to use them directly for preventive purposes in dietary supplements. Efforts are currently being made to prepare dietary supplements enriched with GLS and/or Myr. In this study, we report a simple but efficient chromatographic procedure for the isolation and purification of GLSs from MeOH extract from hoary cress based on a combination of ion exchange and gel permeation chromatography on DEAE-Sephadex A-25 and Sephadex LH-20. To obtain the Myr required for efficient hydrolysis of GLSs into antibacterial ITCs, we developed a rapid method for its extraction from the seeds of Lepidium sativum (garden cress). The yields of GLSs were 22.9 ± 1.2 mg GRN (purity 96%) and 10.4 ± 1.1 mg SBN (purity 92%) from 1 g of dry plant material. Both purified GLSs were used as substrates for the Myr. Analysis of the composition of hydrolysis products (HPs) revealed differences in their hydrolysis rates and in the degree of conversion from GLSs to individual ITCs catalyzed by Myr. When GRNs were cleaved, SFNs were formed in an equimolar ratio, but the formation of pHBITCs was only half that of cleaved SBNs. The decrease in pHBITC content is due to its instability compared to SFN. While SFN is stable in aqueous media during the measurement, pHBITC undergoes non-enzymatic hydrolysis to p-hydroxybenzyl alcohol and thiocyanate ions. Testing of the antimicrobial effects of the HPs formed from GRN by Myr under premix or in situ conditions showed inhibition of the growth of model prokaryotic and eukaryotic microorganisms. This observation could serve as the jumping-off point for the design of a two-component mixture, based on purified GLSs and Myr that is, usable in food or the pharmaceutical industry in the future. [ABSTRACT FROM AUTHOR]

Sulforaphane is considered the bioactive metabolite of glucoraphanin after dietary consumption of broccoli sprouts. Although both molecules pass through the gut lumen to the large intestine in stable form, their biological impact on the first intestinal tract is poorly described. In celiac patients, the function of the small intestine is affected by celiac disease (CD), whose severe outcomes are controlled by gluten-free dietary protocols. Nevertheless, pathological signs of inflammation and oxidative stress may persist. The aim of this study was to compare the biological activity of sulforaphane with its precursor glucoraphanin in a cellular model of gliadin-induced inflammation. Human intestinal epithelial cells (CaCo-2) were stimulated with a pro-inflammatory combination of cytokines (IFN-γ, IL-1β) and in-vitro-digested gliadin, while oxidative stress was induced by H2O2. LC-MS/MS analysis confirmed that sulforaphane from broccoli sprouts was stable after simulated gastrointestinal digestion. It inhibited the release of all chemokines selected as inflammatory read-outs, with a more potent effect against MCP-1 (IC50 = 7.81 µM). On the contrary, glucoraphanin (50 µM) was inactive. The molecules were unable to counteract the oxidative damage to DNA (γ-H2AX) and catalase levels; however, the activity of NF-κB and Nrf-2 was modulated by both molecules. The impact on epithelial permeability (TEER) was also evaluated in a Transwell® model. In the context of a pro-inflammatory combination including gliadin, TEER values were recovered by neither sulforaphane nor glucoraphanin. Conversely, in the context of co-culture with activated macrophages (THP-1), sulforaphane inhibited the release of MCP-1 (IC50 = 20.60 µM) and IL-1β (IC50 = 1.50 µM) only, but both molecules restored epithelial integrity at 50 µM. Our work suggests that glucoraphanin should not merely be considered as just an inert precursor at the small intestine level, thus suggesting a potential interest in the framework of CD. Its biological activity might imply, at least in part, molecular mechanisms different from sulforaphane.

Glucoraphanin is an important glucosinolate which is widely distributed in Brassica vegetables and poses an anticancer effect to humans. Although researchers have paid a lot of attention to the changes in glucoraphanin concentration in seedlings of broccoli over 1–2 weeks, there has been little research focusing on the total whole-sprout glucoraphanin content within broccoli seedlings over 1–5 weeks. However, it is necessary to clarify the changes in total glucoraphanin content during the broccoli sprouting stage as broccoli seedlings are novel plant foods. This research explored glucoraphanin absolute accumulation and the biosynthesis mechanism in broccoli seedlings during a 5-week growth period. The results showed that glucoraphanin accumulation content was higher at week 4 than in the seeds. Moreover, the relative DL-methionine contents increased significantly after 3 weeks. Glucoraphanin synthetic gene expression levels were increased after 3 weeks, but the gene expressions of AOP3 (encoding 2-oxoglutarate-dependent dioxygenases) and MYR (encoding myrosinase) were significantly decreased. Furthermore, the 20 essential DEGs obtained can provide new insight into understanding the developmental regulation of broccoli seedlings. In addition, the results can also provide information on how to obtain higher glucoraphanin contents in broccoli sprouts. [ABSTRACT FROM AUTHOR]

To investigate the mechanism of melatonin (MT)-mediated glutathione (GSH) in promoting glucoraphanin (GRA) and sulforaphane (SF) synthesis, the gene expression pattern and protein content of hairy broccoli roots under MT treatment were analyzed by a combination of RNA-seq and tandem mass spectrometry tagging (TMT) techniques in this study. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that both proteins and mRNAs with the same expression trend were enriched in the "Glutathione metabolism (ko00480)" and "Proteasome (ko03050)" pathways, and most of the differentially expressed genes (DEGs) and differentially abundant proteins (DAPs) regulating the two pathways were downregulated. The results showed that endogenous GSH concentration and GR activity were increased in hairy roots after MT treatment. Exogenous GSH could promote the biosynthesis of GRA and SF, and both exogenous MT and GSH could upregulate the expression of the GSTF11 gene related to the sulfur transport gene, thus promoting the biosynthesis of GRA. Taken together, this study provides a new perspective to explore the complex molecular mechanisms of improving GRA and SF synthesis levels by MT and GSH regulation. [ABSTRACT FROM AUTHOR]

Genetic and pharmacological activation of the transcription factor nuclear factor, erythroid derived 2, like 2 (Nrf2) alleviates high-fat diet (HFD)-induced obesity in mice; however, synthetic Nrf2 activators are not clinically available due to safety concerns. Dietary glucoraphanin (GR), a naturally occurring compound found in cruciferous vegetables that activates Nrf2 and induces its target antioxidant genes. We previously demonstrated that GR increased thermogenesis and mitigated HFD-induced obesity in lean healthy mice. In this study, we investigated the therapeutic effects of GR on pre-existing obesity and associated metabolic disorders, such as hepatic steatosis, with or without low-fat dietary intervention. Eight-week-old male C57BL/6J mice were fed an HFD for 9 weeks to induce obesity. Subsequently, these obese mice were fed either the HFD or a normal chow diet, supplemented with or without GR, for an additional 11 weeks. GR supplementation did not decrease the body weight of HFD-fed mice; however, it significantly reduced plasma alanine aminotransferase and aspartate aminotransferase levels and hepatic triglyceride accumulation. These improvements in liver damage by GR were associated with decreased expression levels of fatty acid synthesis genes and proinflammatory chemokine genes, suppressed c-Jun N-terminal kinase activation, and reduced proinflammatory phenotypes of macrophages in the liver. Moreover, metabolome analysis identified increased hepatic levels of adenosine 5′-monophosphate (AMP) in HFD-GR mice compared with those in HFD mice, which agreed with increased phosphorylation levels of AMP-activated protein kinase. Our results show that GR may have a therapeutic potential for treating obesity-associated hepatic steatosis. [ABSTRACT FROM AUTHOR]

Objectives: This paper presents a systematic review aimed at assessing the therapeutic potential of sulforaphane (SFN) in the treatment of diverse cancer types. Methods: Following Cochrane guidelines for systematic reviews, we conducted an exhaustive search of electronic databases up to May 12, 2023, encompassing PubMed, Cochrane, Embase, Web of Science, Google Scholar, Natural Medicines, ProQuest, ClinicalTrials.gov, and ICTRP. Studies were included if they were human-based RCTs involving cancer patients where SFN was the primary experimental treatment. The Cochrane Risk of Bias tool for RCTs (RoB2) was used for quality assessment. Results: Eight studies investigating the efficacy and safety of SFN in prostate cancer (PCa), breast cancer, pancreatic cancer, andmelanoma were identified and included in the review. The dosing regimens were variable and inconsistent across the studies. SFN treatment led to statistically significant alterations in several vital genes and histological biomarkers across the studies. However, it did not impact some other key genes. Although not statistically significant, SFN improved overall survival in pancreatic cancer patients. The results on prostate-specific antigen (PSA) were inconsistent in PCa. None of the studies reported significant differences between SFN and comparative controls in terms of adverse events. Conclusion: SFN has emerged as a promising and safe therapeutic agent for diverse cancer types. Nevertheless, the high levels of methodological and clinical heterogeneity across the included studies precluded the possibility of conducting meta-analyses. Further robust clinical investigations to conclusively ascertain the chemotherapeutic potential of SFN in the management of various cancer forms are needed. [ABSTRACT FROM AUTHOR]

Glucosinolates (GSLs), crucial secondary metabolites in cruciferous vegetables, hydrolyze upon consumption or mechanical damage, forming bioactive compounds with anti-cancer properties, such as glucoraphanin (GRA). Among cruciferous vegetables, broccoli stands out for its high GSL content, which varies significantly among different genotypes. This study aimed to characterize and quantify glucosinolate compounds in broccoli using LC-HRMS2 and UPLC. We identified thirteen GSLs in 191 broccoli genotypes, including seven aliphatic, five indole, and one aromatic glucosinolate. The GSL content in these genotypes ranged from 0.1705 to 5.8174 mg/g (DW). We also explored GSL diversity and content in seven developmental organs, finding high diversity and content in seedling roots and florets. Notably, genotype No. 300 had the highest GSL content (5.8174 mg/g, DW) and GRA (3.1545 mg/g, DW), along with a larger flower bulb diameter (13.4 cm) and a shorter growth stage (11 days), demonstrating its potential for breeding GRA-rich broccoli. To our knowledge, this study encompasses the largest number of broccoli genotypes to date, broadening our understanding of GSLs' diversity and content in broccoli. These findings may provide valuable resources for future breeding or the commercial cultivation of GRA-rich broccoli. [ABSTRACT FROM AUTHOR]

Broccoli is a rich source of diverse bioactive compounds, but how their contents are influenced by different growing seasons and variations in broccoli head sizes remains elusive. To address this question, we quantified sixteen known bioactive compounds and seven minerals in broccoli with varying head sizes obtained in two different growing seasons. Our results suggest that the contents of vitamin C, total phenols, carotenoids, and glucoraphanin were significantly higher in samples from the summer–autumn season, showing increases of 157.46%, 34.74%, 51.80%, and 17.78%, respectively, compared with those from the winter–spring season. Moreover, chlorogenic acid is a phenolic compound with relatively high contents among the six detected, while beta-sitosterol is the sterol with relatively high contents. Further, principal component analysis was conducted to rank the comprehensive scores of the profiles of phenolic compounds, phytosterols, and minerals, demonstrating that the broccoli samples grown during the summer–autumn season achieved the highest composite scores. Our results indicate that broccoli heads from the summer–autumn season are richer in a combination of bioactive compounds and minerals than those from the winter–spring season based on the composite score. This study extends our understanding of the nutrition profiles in broccoli and also lays the foundation for breeding broccoli varieties with improved nutrition quality.

ABSTRACT Crohn’s disease (CD) is a presentation of inflammatory bowel disease (IBD) that manifests in childhood and adolescence and involves chronic and severe enterocolitis, immune and gut microbial dysregulation, and other complications. Diet and gut-microbiota-produced metabolites are sources of anti-inflammatories that could ameliorate symptoms. However, questions remain on how IBD influences biogeographic patterns of microbial location and function in the gut, how early life transitional gut communities are affected by IBD and diet interventions, and how disruption to biogeography alters disease mediation by diet components or microbial metabolites. Many studies on diet and IBD use a chemically induced ulcerative colitis model, despite the availability of an immune-modulated CD model. Interleukin-10-knockout (IL-10-KO) mice on a C57BL/6 background, beginning at age 4 or 7 weeks, were fed a control diet or one containing 10% (wt/wt) raw broccoli sprouts, which was high in the sprout-sourced anti-inflammatory sulforaphane. Diets began 7 days prior to, and for 2 weeks after inoculation with Helicobacter hepaticus, which triggers Crohn’s-like symptoms in these immune-impaired mice. The broccoli sprout diet increased sulforaphane in plasma; decreased weight stagnation, fecal blood, and diarrhea associated; and increased microbiota richness in the gut, especially in younger mice. Sprout diets resulted in some anatomically specific bacteria in younger mice and reduced the prevalence and abundance of pathobiont bacteria which trigger inflammation in the IL-10-KO mouse, for example, Escherichia coli and Helicobacter. Overall, the IL-10-KO mouse model is responsive to a raw broccoli sprout diet and represents an opportunity for more diet-host-microbiome research.IMPORTANCETo our knowledge, IL-10-KO mice have not previously been used to investigate the interactions of host, microbiota, and broccoli, broccoli sprouts, or broccoli bioactives in resolving symptoms of CD. We showed that a diet containing 10% raw broccoli sprouts increased the plasma concentration of the anti-inflammatory compound sulforaphane and protected mice to varying degrees against disease symptoms, including weight loss or stagnation, fecal blood, and diarrhea. Younger mice responded more strongly to the diet, further reducing symptoms, as well as increased gut bacterial richness, increased bacterial community similarity to each other, and more location-specific communities than older mice on the diet intervention. Crohn’s disease disrupts the lives of patients and requires people to alter dietary and lifestyle habits to manage symptoms. The current medical treatment is expensive with significant side effects, and a dietary intervention represents an affordable, accessible, and simple strategy to reduce the burden of symptoms.

Background and aimsObservational data indicate that diets rich in fruits and vegetables have a positive effect on inflammatory status, improve metabolic resilience and may protect against the development of non-communicable diseases. Nevertheless, experimental evidence demonstrating a causal relationship between nutrient intake (especially whole foods) and changes in metabolic health is scarce. This study investigated the pleiotropic effects of sulforaphane from broccoli sprouts, compared to pea sprouts, on biomarkers of endothelial function, inflammation and metabolic stress in healthy participants subjected to a standardized caloric challenge.MethodsIn this double-blind, crossover, randomized, placebo-controlled trial 12 healthy participants were administered 16 g broccoli sprouts, or pea sprouts (placebo) followed by the standardized high-caloric drink PhenFlex given to disturb healthy homeostasis. Levels of inflammatory biomarkers and metabolic parameters were measured in plasma before and 2 h after the caloric overload.ResultsAdministration of broccoli sprouts promoted an increase in levels of CCL-2 induced by caloric load (p = 0.017). Other biomarkers (sICAM-1, sVCAM-1, hs-CRP, and IL-10) individually showed insignificant tendencies toward increase with administration of sulforaphane. Combining all studied biomarkers into the systemic low-grade inflammation score further confirmed upregulation of the inflammatory activity (p = 0.087) after sulforaphane. No significant effects on biomarkers of metabolic stress were detected.ConclusionThis study has demonstrated that sulforaphane facilitated development of a mild pro-inflammatory state during the caloric challenge, which could be suggestive of the onset of the hormetic response induced by this phytonutrient. The use of integrative outcomes measures such as the systemic low-grade inflammation score can be viewed as a more robust approach to study the subtle and pleiotropic effects of phytonutrients.Clinical trial registration:www.clinicaltrials.gov, identifier NCT05146804.

ObjectivesThis paper presents a systematic review aimed at assessing the therapeutic potential of sulforaphane (SFN) in the treatment of diverse cancer types.MethodsFollowing Cochrane guidelines for systematic reviews, we conducted an exhaustive search of electronic databases up to May 12, 2023, encompassing PubMed, Cochrane, Embase, Web of Science, Google Scholar, Natural Medicines, ProQuest, ClinicalTrials.gov, and ICTRP. Studies were included if they were human-based RCTs involving cancer patients where SFN was the primary experimental treatment. The Cochrane Risk of Bias tool for RCTs (RoB2) was used for quality assessment.ResultsEight studies investigating the efficacy and safety of SFN in prostate cancer (PCa), breast cancer, pancreatic cancer, and melanoma were identified and included in the review. The dosing regimens were variable and inconsistent across the studies. SFN treatment led to statistically significant alterations in several vital genes and histological biomarkers across the studies. However, it did not impact some other key genes. Although not statistically significant, SFN improved overall survival in pancreatic cancer patients. The results on prostate-specific antigen (PSA) were inconsistent in PCa. None of the studies reported significant differences between SFN and comparative controls in terms of adverse events.ConclusionSFN has emerged as a promising and safe therapeutic agent for diverse cancer types. Nevertheless, the high levels of methodological and clinical heterogeneity across the included studies precluded the possibility of conducting meta-analyses. Further robust clinical investigations to conclusively ascertain the chemotherapeutic potential of SFN in the management of various cancer forms are needed.Systematic review registrationhttps://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42022323788, identifier CRD42022323788.

The isothiocyanate's chemoprevention properties are reported to be present in cruciferous veggies through a variety of mechanisms. Sulforaphane, a phytochemical found in green leafy vegetables, has shown promise in the prevention and treatment of several cancers, including those of the prostate, breast, colon, skin, urinary bladder, and oral cavities. These malignancies include those that affect these organs. This substance is naturally present in broccoli sprouts, kale, cabbage, cauliflower, and garden cress. Broccoli should be a regular part of your diet because it contains a variety of bioactive substances such as vitamins, polyphenols, sulfides, glucosinolates, and antioxidants. Sulforaphane may be used as an inexpensive replacement or dietary supplement for chemo preventive therapy, according to the findings of epidemiological and experimental studies. Phase 2 detoxification enzymes like glutathione transferases, epoxide hydrolase, NAD(P)H: quinone reductase, and glucuronosyltransferases, as well as epoxide hydrolase and epoxide hydrolase, are produced when the body is stimulated. This is a useful tactic for preventing cancer and fending off the harm that electrophiles and reactive oxygen species can cause. Isothiocyanates are widely distributed in the Cruciferae family and Brassica genus of food plants, which include both broccoli and cauliflower. The most typical form of these substances is glucosinolate precursors. Sulforaphane and 4-methylsulfinylbutyl isothiocyanate, two of these isothiocyanates, are particularly powerful inducers of phase 2 enzymes. It is feasible to successfully extract glucosinolates and isothiocyanates from plants by homogenizing them at a temperature of around 50 degrees Celsius in a solution of equal parts dimethyl sulfoxide, dimethylformamide, and acetonitrile. This method avoids the hydrolysis of glucosinolates by myrosinase. It's interesting to note that glucoraphanin, the precursor to sulforaphane, is 10-100 times more abundant in 3-day-old sprouts of various cruciferous vegetables, including broccoli and cauliflower than it is in fully grown veggies. Dimethylbenz(a)anthracene-treated rats displayed notable reductions in mammary tumor occurrence, quantity, and rate of growth when fed extracts from 3-day-old broccoli sprouts. These extracts' primary enzyme-inducing components were either glucoraphanin or sulforaphane. As a result, crucifer sprouts might provide an equivalent amount of cancer prevention to eating far larger quantities of the same mature vegetable species. [ABSTRACT FROM AUTHOR]

Lepidium draba (hoary cress) is a perennial plant belonging to the Brassicaceae family that produces two dominant glucosinolates (GLSs): glucoraphanin (GRN) and sinalbin (SBN). They represent the stored form, which is converted upon the myrosinase (Myr) hydrolysis activity to active compounds, mainly isothiocyanates (ITCs) such as sulforaphane (SFN) or p-hydroxybenzyl isothiocyanate (pHBITC). Research on ITCs that have proven anticancer, antimicrobial, and chemoprotective properties is usually conducted with pure commercially available compounds. However, these are chemically reactive, making it difficult to use them directly for preventive purposes in dietary supplements. Efforts are currently being made to prepare dietary supplements enriched with GLS and/or Myr. In this study, we report a simple but efficient chromatographic procedure for the isolation and purification of GLSs from MeOH extract from hoary cress based on a combination of ion exchange and gel permeation chromatography on DEAE-Sephadex A-25 and Sephadex LH-20. To obtain the Myr required for efficient hydrolysis of GLSs into antibacterial ITCs, we developed a rapid method for its extraction from the seeds of Lepidium sativum (garden cress). The yields of GLSs were 22.9 ± 1.2 mg GRN (purity 96%) and 10.4 ± 1.1 mg SBN (purity 92%) from 1 g of dry plant material. Both purified GLSs were used as substrates for the Myr. Analysis of the composition of hydrolysis products (HPs) revealed differences in their hydrolysis rates and in the degree of conversion from GLSs to individual ITCs catalyzed by Myr. When GRNs were cleaved, SFNs were formed in an equimolar ratio, but the formation of pHBITCs was only half that of cleaved SBNs. The decrease in pHBITC content is due to its instability compared to SFN. While SFN is stable in aqueous media during the measurement, pHBITC undergoes non-enzymatic hydrolysis to p-hydroxybenzyl alcohol and thiocyanate ions. Testing of the antimicrobial effects of the HPs formed from GRN by Myr under premix or in situ conditions showed inhibition of the growth of model prokaryotic and eukaryotic microorganisms. This observation could serve as the jumping-off point for the design of a two-component mixture, based on purified GLSs and Myr that is, usable in food or the pharmaceutical industry in the future.

ABSTRACT Inflammatory bowel diseases (IBDs) are devastating conditions of the gastrointestinal tract with limited treatments, and dietary intervention may be effective and affordable for managing symptoms. Glucosinolate compounds are highly concentrated in broccoli sprouts, especially glucoraphanin (GLR), and can be metabolized by certain mammalian gut bacteria into anti-inflammatory isothiocyanates, such as sulforaphane. Gut microbiota exhibit biogeographic patterns, but it is unknown if colitis alters these or whether the location of glucoraphanin-metabolizing bacteria affects anti-inflammatory benefits. We fed specific pathogen-free C57BL/6 mice either a control diet or a 10% steamed broccoli sprout diet and gave a three-cycle regimen of 2.5% dextran sodium sulfate (DSS) in drinking water over a 34-day experiment to simulate chronic, relapsing ulcerative colitis (UC). We monitored body weight, fecal characteristics, lipocalin, serum cytokines, and bacterial communities from the luminal- and mucosal-associated populations in the jejunum, cecum, and colon. Mice fed the broccoli sprout diet with DSS treatment performed better than mice fed the control diet with DSS, and had significantly more weight gain, lower Disease Activity Index scores, lower plasma lipocalin and proinflammatory cytokines, and higher bacterial richness in all gut locations. Bacterial communities were assorted by gut location but were more homogenous across locations in the control diet + DSS mice. Importantly, our results showed that broccoli sprout feeding abrogated the effects of DSS on gut microbiota, as bacterial richness and biogeography were similar between mice receiving broccoli sprouts with and without DSS. Collectively, these results support the protective effect of steamed broccoli sprouts against dysbiosis and colitis induced by DSS. IMPORTANCE Evaluating bacterial communities across different locations in the gut provides a greater insight than fecal samples alone and provides an additional metric by which to evaluate beneficial host-microbe interactions. Here, we show that 10% steamed broccoli sprouts in the diet protects mice from the negative effects of dextran sodium sulfate-induced colitis, that colitis erases biogeographic patterns of bacterial communities in the gut, and that the cecum is not likely to be a significant contributor to colonic bacteria of interest in the DSS mouse model of ulcerative colitis. Mice fed the broccoli sprout diet during colitis performed better than mice fed the control diet while receiving DSS. The identification of accessible dietary components and concentrations that help maintain and correct the gut microbiome may provide universal and equitable approaches to IBD prevention and recovery, and broccoli sprouts represent a promising strategy.

Glucosinolates (GSLs), precursors of isothiocyanates (ITCs), are present in Brassicaceae plants have been found to have health benefits. Sulforaphane (4-(methylsulfinyl)butyl ITC) is an ITC stored in the formof 4-(methylsulfinyl)butyl GSL (glucoraphanin, 4MSOB) in Brassica vegetables, such as broccoli and kale. Sulforaphane activates Nrf2 expression, a transcription factor responsible for inducing physiological activities such as detoxification in the human body, and it represents a functional component unique to cruciferous vegetables. Raphanobrassica is an inter-generic hybrid between radish and kale, and it contains a high amount of 4MSOB. However, Raphanobrassica contains as much 4-methylsulfinyl-3-butenyl GSL (glucoraphenin, 4MSO3B) as it does 4MSOB. GLUCORAPHASATIN SYNTHASE 1 (GRS1) is an enzyme present in radish that synthesizes 4-methylthio-3-butenyl GSL (glucoraphasatin, 4MT3B), a precursor of 4MSO3B, using 4-(methylthio)butyl GSL (glucoerucin, 4MTB) as a substrate. Since the precursor of 4MSOB is also 4MTB, it was considered that both 4MSOB and 4MSO3B accumulate owing to competition in Raphanobrassica. We hypothesized that owing to the impaired function of GRS1 in Raphanobrassica, it may be possible to breed Raphanobrassica cultivars containing a high 4MSOB content. In this study, we generated Raphanobrassica populations with functional and defective GRS1 and compared the GSL composition in the two populations using high-performance liquid chromatography. The mean 4MSOB content in leaves of the defective-type populations was higher than that in the functionaltype population, and the defective/functional ratio ranged from 2.02 to 2.51-fold, supporting this hypothesis. Furthermore, leaves, flower buds, stems, and roots contained higher amounts of 4MSOB in the defective population than in the functional population. The leaf 4MSOB content of defective Raphanobrassica grown in this study was comparable to that of previously studied vegetables (such as broccoli sprouts) with high 4MSOB content. Raphanobrassica with defective GRS1 represents a new leafy vegetable with high 4MSOB content which exhibits anti-cancerous and anti-inflammatory potentials. [ABSTRACT FROM AUTHOR]

Background: The Purpose of this study was to investigate the effects of sulforaphane glucosinolate (SGS) from broccoli seed extracts on immune function and common cold symptoms in healthy Japanese adults. Methods: This randomized, placebo-controlled, double-blind, parallel-group comparison study was conducted on Japanese adults who had worse health-related quality of life on the SF-8 and were more likely to catch colds from October 22, 2020 to April 2, 2021. Individuals who agreed to participate in the study were randomly assigned to either the SGS 100 mg/day group or the placebo group (n = 33 each). The intervention lasted 8 weeks. The outcomes of this study were the cumulative and the maximum number of days for which common cold symptoms persisted during the intervention period per subject (primary), and the frequency with which subjects experienced these symptoms per group (secondary). Additionally, the incidence rate of subjects infected with the influenza virus, immune indices, and SF-8 were assessed. Results: Based on a per protocol set, 64 subjects (33 in the SGS group and 31 in the placebo group) were analyzed. After the 8-week interventional period, the cumulative number of days for which common cold symptoms per subject was significantly lower for the SGS group than the placebo group (12.1 ± 13.5 for the SGS group and 20.2 ± 18.2 for the placebo group). The cumulative number of days of common cold symptoms per group was also significantly lower in the SGS group than in the placebo group (400 days in the SGS group and 626 days in the placebo group). No changes attributable to the SGS intervention were observed in the other outcomes. There were no adverse events due to food ingestion. Conclusion: A period of 8 weeks of SGS intake was shown to alleviate the onset of common cold symptoms in healthy Japanese adults who were prone to catching colds and had a low subjective sense of wellness. [ABSTRACT FROM AUTHOR]

Abstract Eruca vesicaria subsp. sativa is a leafy vegetable of the Brassicaceae family known for its pungency. Variation in growing conditions, leaf age, agronomic practices, and variety choice lead to inconsistent quality, especially in content of isothiocyanates (ITCs) and their precursor glucosinolates (GSLs). We present the first linkage and Quantitative Trait Loci (QTL) map for Eruca, generated using a population of 139 F4 lines. A significant environmental effect on the abundance of primary and secondary metabolites was observed, with UK-grown plants containing significantly higher concentrations of glucoraphanin, malic acid, and total sugars. Italian-grown plants were characterized by higher concentrations of glucoerucin, indolic GSLs, and low monosaccharides. 20 QTL were identified and associated with robust SNP markers. Five genes putatively associated with the synthesis of the GSL 4-methoxyglucobrassicin (4MGB) were identified as candidate regulators underlying QTL. Analysis revealed that orthologs of MYB51, IGMT1 and IGMT4 present on LG1 are associated with 4MGB concentrations in Eruca. This research illustrates the utility of the map for identifying genes associated with nutritional composition in Eruca and its value as a genetic resource to assist breeding programs for this leafy vegetable crop.

Abstract Glucoraphanin (GRA) is present in the seeds and nutrient organs of broccoli and is the precursor of the anti-cancer compound sulforaphane (SF). The hairy roots obtained by infecting broccoli (Brassica oleracea L. var. Italic Planch) leaves with Agrobacterium rhizogenes (ATCC15834) are phytohormonally autonomous, genetically stable, and can produce large amounts of the anti-cancer substance SF. Melatonin (MT) is a natural hormone widely found in plants. Studies have shown that melatonin can regulate the synthesis of secondary metabolites of downstream targets by mediating the synthesis of signal molecules. However, whether MT regulates the synthesis of NO and H2O2 and mediates the synthesis mechanism of secondary metabolites, GRA and SF, is not yet clear. In this study, the hairy roots of broccoli were treated with 500 μmol/L MT, and the genome of broccoli (Brassica oleracea L. var. botrytis L) was used as the reference genome for transcriptome analysis. By this approach, we found that MT regulates the synthesis of NO and H2O2 and mediates the synthesis of secondary metabolites GRA and SF. GO annotations indicated that DEGs involved in the MT treatment of broccoli hairy roots were mainly related to catalytic activity, cells, and metabolic processes; the KEGG pathway analysis indicated that MT treatment likely affects the hormone signal transduction process in broccoli hairy roots; broccoli hairy roots were treated with 500 μmol/L MT for 0, 6, 12, 20, and 32 h, respectively; compared with 0 h, the yield of GRA and SF increased under the other treatments. The highest yields of GRA and SF occurred at 12 h. The NO content was the highest at 12 h, and the H2O2 content was positively correlated with MT concentration. The content of NO and H2O2 were regulated, and the content of GRA and SF was increased under MT treatment. NO synthase inhibitor (L-NAME and TUN) could effectively inhibit the content of NO in broccoli hairy roots and reduce GRA and SF yield; MT could regulate NO levels by regulating NO synthesis-related enzymes and could alleviate the reduction of NO content in tissue cells caused by NO synthase inhibitor and promote NO synthesis. These results have important theoretical implications for understanding the regulation of GRA and SF synthesis events by NO and H2O2.

BACKGROUND: Broccoli has received widespread attention because of its anti‐inflammatory and antioxidant effects. The present study aimed to explore the composition of broccoli seed extract (BSE) and its effect on colitis induced by dextran sulfate sodium (DSS). RESULTS: BSE mainly comprises glucoraphanin and polysaccharides composed of arabinose, galactose, glucose and mannose. Animal experiments suggested that BSE intervention effectively reversed body weight loss, suppressed the levels of proinflammatory interleukin‐6, tumor necrosis factor‐α and interleukin‐1β, and elevated the levels of anti‐inflammatory interleukin‐10 and the activities of superoxide dismutase and glutathione in DSS‐induced colitis mice. According to histopathologic and immunohistochemical analysis of colon tissue, BSE intervention may repair the intestinal barrier by upregulating mRNA levels and the expression of tight junction proteins (claudin‐1, occludin and zonula occludens‐1). Gas chromatography–mass spectrometry (MS) analysis demonstrated that cecal short‐chain fatty acids in mice with BSE administration were significantly increased compared with the model group. Sulforaphane and sulforaphane‐N‐acetylcysteine were only detected in BSE group mice by ultra‐performance liquid chromatography‐MS analysis. In addition, BSE intervention evidently increased the abundance of Alistipeds, Coriobacteriaceae UCG‐002 and Bifidobacterium and decreased the abundance of Escheichia‐Shinella, Lachnospiraceae others, Parabacteroides, Ruminococcaceae others and Turicibacter, which possibly promoted carbohydrate metabolism and short‐chain fatty acid production. CONCLUSION: The present study aimed to elucidate the effect of BSE on colitis and found that BSE, as a novel food ingredient, has great potential for the improvement of colitis. © 2022 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Myrosinase is a key tool for the fast and efficient preparation of sulforaphane which is one of the prominent natural ingredients found in brassicaceous vegetables. Here, the glucoraphanin-hydrolyzing activity of a Yarrowia lipolytica 20–8 harboring myrosinase reached 73.28 U/g dry cell weight, indicating that it had a potential application in sulforaphane preparation from glucoraphanin. An efficient and reusable process for sulforaphane preparation via myrosinase produced by Y. lipolytica 20–8 was constructed. In detail, as high as 10.32 mg sulforaphane could be produced from 1 g broccoli seed under the reaction of 40 U yeast whole-cell catalyst within 15 min with the conversion efficiency of 99.86%. Moreover, when the yeast whole-cell catalyst was reused 7 and 10 times, as high as 92.53% and 87.56% of sulforaphene yield of the initial level could be retained, respectively. Therefore, this yeast whole-cell is a potent biocatalyst for the efficient and reusable preparation of sulforaphane.

Glucoraphanin is an important glucosinolate which is widely distributed in Brassica vegetables and poses an anticancer effect to humans. Although researchers have paid a lot of attention to the changes in glucoraphanin concentration in seedlings of broccoli over 1–2 weeks, there has been little research focusing on the total whole-sprout glucoraphanin content within broccoli seedlings over 1–5 weeks. However, it is necessary to clarify the changes in total glucoraphanin content during the broccoli sprouting stage as broccoli seedlings are novel plant foods. This research explored glucoraphanin absolute accumulation and the biosynthesis mechanism in broccoli seedlings during a 5-week growth period. The results showed that glucoraphanin accumulation content was higher at week 4 than in the seeds. Moreover, the relative DL-methionine contents increased significantly after 3 weeks. Glucoraphanin synthetic gene expression levels were increased after 3 weeks, but the gene expressions of AOP3 (encoding 2-oxoglutarate-dependent dioxygenases) and MYR (encoding myrosinase) were significantly decreased. Furthermore, the 20 essential DEGs obtained can provide new insight into understanding the developmental regulation of broccoli seedlings. In addition, the results can also provide information on how to obtain higher glucoraphanin contents in broccoli sprouts.

To investigate the mechanism of melatonin (MT)-mediated glutathione (GSH) in promoting glucoraphanin (GRA) and sulforaphane (SF) synthesis, the gene expression pattern and protein content of hairy broccoli roots under MT treatment were analyzed by a combination of RNA-seq and tandem mass spectrometry tagging (TMT) techniques in this study. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that both proteins and mRNAs with the same expression trend were enriched in the “Glutathione metabolism (ko00480)” and “Proteasome (ko03050)” pathways, and most of the differentially expressed genes (DEGs) and differentially abundant proteins (DAPs) regulating the two pathways were downregulated. The results showed that endogenous GSH concentration and GR activity were increased in hairy roots after MT treatment. Exogenous GSH could promote the biosynthesis of GRA and SF, and both exogenous MT and GSH could upregulate the expression of the GSTF11 gene related to the sulfur transport gene, thus promoting the biosynthesis of GRA. Taken together, this study provides a new perspective to explore the complex molecular mechanisms of improving GRA and SF synthesis levels by MT and GSH regulation.

Glucosinolates (GSLs), crucial secondary metabolites in cruciferous vegetables, hydrolyze upon consumption or mechanical damage, forming bioactive compounds with anti-cancer properties, such as glucoraphanin (GRA). Among cruciferous vegetables, broccoli stands out for its high GSL content, which varies significantly among different genotypes. This study aimed to characterize and quantify glucosinolate compounds in broccoli using LC-HRMS2 and UPLC. We identified thirteen GSLs in 191 broccoli genotypes, including seven aliphatic, five indole, and one aromatic glucosinolate. The GSL content in these genotypes ranged from 0.1705 to 5.8174 mg/g (DW). We also explored GSL diversity and content in seven developmental organs, finding high diversity and content in seedling roots and florets. Notably, genotype No. 300 had the highest GSL content (5.8174 mg/g, DW) and GRA (3.1545 mg/g, DW), along with a larger flower bulb diameter (13.4 cm) and a shorter growth stage (11 days), demonstrating its potential for breeding GRA-rich broccoli. To our knowledge, this study encompasses the largest number of broccoli genotypes to date, broadening our understanding of GSLs’ diversity and content in broccoli. These findings may provide valuable resources for future breeding or the commercial cultivation of GRA-rich broccoli.

As the prevalence of inflammatory bowel diseases (IBD) rises, the etiology of IBD draws increasing attention. Glucoraphanin (GRP), enriched in cruciferous vegetables, is a precursor of sulforaphane, known to have anti-inflammatory and antioxidative effects. We hypothesized that dietary GRP supplementation can prevent mitochondrial dysfunction and oxidative stress in an acute colitis mouse model induced by dextran sulfate sodium (DSS). Eight-week-old mice were fed a regular rodent diet either supplemented with or without GRP. After 4 weeks of dietary treatments, half of the mice within each dietary group were subjected to 2.5% DSS treatment to induce colitis. Dietary GRP decreased DSS-induced body weight loss, disease activity index, and colon shortening. Glucoraphanin supplementation protected the colonic histological structure, suppressed inflammatory cytokines, interleukin (IL)-1β, IL-18, and tumor necrosis factor-α (TNF-α), and reduced macrophage infiltration in colonic tissues. Consistently, dietary GRP activated AMP-activated protein kinase (AMPK), peroxisome proliferator-activated receptor-gamma coactivator (PGC)-1α, and nuclear factor erythroid 2-related factor 2 (NRF2) pathways in the colonic tissues of DSS-treated mice, which was associated with increased mitochondrial DNA and decreased content of the oxidative product 8-hydroxydeoxyguanosine (8-OHDG), a nucleotide oxidative product of DNA. In conclusion, dietary GRP attenuated mitochondrial dysfunction, inflammatory response, and oxidative stress induced by DSS, suggesting that dietary GRP provides a dietary strategy to alleviate IBD symptoms. [ABSTRACT FROM AUTHOR]

Hairy roots obtained by infecting broccoli (Brassica oleracea var. italica) leaves with Agrobacterium rhizogenes (ATCC15834) have the characteristics of phytohormone autonomy, genetic stability and can produce a large amount of the anti-cancer substance Sulforaphane (SF) and the biosynthetic precursor Glucoraphanin (GRA). Under the induction of the exogenous signaling molecule methyl jasmonate (MeJA), the production of SF in broccoli hairy roots was significantly increased. However, the molecular mechanism of GRA and SF synthesis in hairy roots of broccoli treated with MeJA has not been reported. In this study, according to the yield of GRA and SF, the best concentration of MeJA treatment for hairy roots of broccoli was selected. After 18 days of growth, broccoli hairy roots were treated with 10 mmol L–1 MeJA for 0, 3, 6, 9 and 12 h. Compared with 0 h, the yield of GRA and SF increased under other treatments. The highest yield of GRA and SF occurred at 9 h, which were 2.22-fold and 1.74-fold higher than those at 0 h. Brassica oleracea var. botrytis was used as reference genome, and 5,757 differentially expressed genes (DEG) were observed at 0, 3, 6, 9 and 12 h under 10 mmol L–1 MeJA treatment, of which 4,673 were down-regulated and 1084 were up-regulated. The key genes regulating GRA synthesis, CYP79F1, CYP83A1, UGT74B1, FMOGS-OX5 and GSL-OH, were up-regulated at 0 and 3 h, and down-regulated the rest of the time; BCAT2 was up-regulated at 6, 9, 12 h, and at 0, 3 h expression was down-regulated, transcription factors MYB28 and MYB29 were down-regulated by exogenous MeJA treatment. A pathway of GRA biosynthesis and transformation pathways in MeJA-treated broccoli hairy roots was simulated and the molecular mechanism of GRA biosynthesis and SF accumulation in broccoli hairy roots under MeJA treatment was revealed. [ABSTRACT FROM AUTHOR]

Here, the impact of 300 mg L-1 slightly acidic electrolyzed water ice (SAEWI) on postharvest senescence, glucosinolate and sulforaphane contents, and the expression levels of key genes in the glucosinolate metabolic pathway in broccoli was investigated. Results indicated that SAEWI delayed yellowing senescence by slowing the decline of chlorophyll content. Moreover, SAEWI upregulated the expression of genes involved in aliphatic and indole glucosinolate synthesis. Specifically, after 1 d of shelf life, the SAEWI-treated group exhibited 1.01–1.96 times higher expression levels of BoMYB28 , BoSUR1 , BoST5 , BoMYB51 , BoCYP79B2 , and BoCYP83B1 compared to the control group. Similarly, the expression levels of BoCYP79F1 , BoUGT74B1 , and BoFMO GS-OX2 were 2.51–8.67 times higher in the SAEWI-treated group. Consequently, the SAEWI treatment resulted in elevated concentrations of glucoraphanin, glucoiberin, and 4-Methoxy-glucobrassicin, contributing to an overall increase of more than 40% in the total glucosinolate content. Additionally, the expression level of BoMYR in the SAEWI-treated group exceeded that of the control group after 1 d of shelf life, accompanied by enhanced myrosinase activity. These changes led to higher sulforaphane content in the SAEWI-treated group, which was found to be 2.65–4.33 times higher than that of the control group. In conclusion, the findings suggest that SAEWI treatment in the cold chain circulation model not only delays postharvest quality decline but also boosts the accumulation of active substances in broccoli. • SAEWI slows down postharvest aging of broccoli. • SAEWI upregulates the expression of key genes involved in GLS. • SAEWI increases the content of sulforaphane in postharvest broccoli. [ABSTRACT FROM AUTHOR]

Sulforaphane (SFN), an isothiocyanate derived from glucoraphanin, has antioxidant, and anti-inflammatory effects that may be beneficial for improving liver function. However, few studies regarding the effects of glucoraphanin on the biological markers related to liver function, such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), and gamma-glutamyltransferase (γ-GTP) in healthy individuals have been reported. This randomized, double-blind, placebo-controlled parallel- group trial was conducted from April 22 to December 25, 2021 and compared the effects of broccoli sprout supplements enriched in glucoraphanin (glucoraphanin supplements) (n = 35) with those of placebo supplements (n = 35). This trial was registered with the University Hospital Medical Information Network Clinical Trial Registry (UMIN-CTR; ID number UMIN000044005) https://center6.umin.ac.jp/cgi-open-bin/ctr_e/ctr_view. cgi?recptno=R000050252. Glucoraphanin significantly improved serum ALT levels at 24 weeks compared to placebo supplements. However, no significant difference in serum glutathione levels, one of the major antioxidants synthesized in the liver, was observed between the two groups. In conclusion, daily intake of the glucoraphanin supplements is beneficial for maintaining liver health in healthy, middle-aged adults with high-normal serum hepatic biomarkers, although further studies focusing on other antioxidant markers are needed to understand how glucoraphanin improves liver function.

Myrosinase can hydrolyze glucosinolates to generate isothiocyanates, which have cancer prevention and anti-cancer properties. The main sources of myrosinase are cruciferous plants. To further improve the efficiency of isothiocyanates preparation, it is necessary to explore novel sources of myrosinases. In this study, we described a bacterium, Shewanella baltica Myr-37, isolated from marine mud, capable of producing a novel myrosinase (Smyr37) with a molecular weight of 100 kDa. The crude enzyme of Smyr37 showed the highest activity at 50 °C and pH 8.0. The sinigrin- and glucoraphanin-hydrolyzing activities of Smyr37 were 6.95 and 5.87 U/mg, respectively. Moreover, when the reaction temperature was 40 °C and pH was 7.0, the crude enzyme of Smyr37 could efficiently degrade glucoraphanin into sulforaphane within 25 min with a yield of 0.57 mg/mL. The corresponding conversion efficiency of sulforaphane from glucoraphanin was 89%. In summary, S. baltica Myr-37 myrosinase Smyr37, a novel myrosinase, can be used in the preparation of isothiocyanates. [ABSTRACT FROM AUTHOR]

Glucoraphanin (GRA) is a natural compound that has shown beneficial effects in chronic diseases and in central nervous system disorders. Moreover, GRA displayed antidepressant activity in preclinical models. We have previously demonstrated that a single intracerebroventricular administration of soluble amyloid-beta 1-42 (sAβ 1-42) in rat evokes a depressive-like phenotype by increasing immobility frequency in the forced swimming test (FST). The aim of this work was to investigate the effect of GRA in naïve and in sAβ-1-42-treated rats by using the FST. Behavioural analyses were accompanied by neurochemical and biochemical measurements in the prefrontal cortex (PFC), such as serotonin (5-HT), noradrenaline (NA), kynurenine (KYN), tryptophan (TRP), reactive oxygen species (ROS) and the transcription nuclear factor kappa B (NF-kB) levels. We reported that GRA administration in naïve rats at the dose of 50 mg/kg reduced the immobility frequency in the FST and increased 5-HT and NA levels in the PFC compared to controls. At the same dose, GRA reverted depressive-like effects of sAβ 1-42 administration, restored the 5-HT levels and reduced NF-kB, KYN and ROS levels in PFC. In conclusion, GRA rapidly reverting depressive-like behaviour, together with biochemical and neurochemical alterations, might represent a safe and natural candidate for the treatment of depression. [ABSTRACT FROM AUTHOR]

Glucoraphanin (GRA) is present in the seeds and nutrient organs of broccoli and is the precursor of the anti-cancer compound sulforaphane (SF). The hairy roots obtained by infecting broccoli (Brassica oleracea L. var. Italic Planch) leaves with Agrobacterium rhizogenes (ATCC15834) are phytohormonally autonomous, genetically stable, and can produce large amounts of the anti-cancer substance SF. Melatonin (MT) is a natural hormone widely found in plants. Studies have shown that melatonin can regulate the synthesis of secondary metabolites of downstream targets by mediating the synthesis of signal molecules. However, whether MT regulates the synthesis of NO and H2O2 and mediates the synthesis mechanism of secondary metabolites, GRA and SF, is not yet clear. In this study, the hairy roots of broccoli were treated with 500 μmol/L MT, and the genome of broccoli (Brassica oleracea L. var. botrytis L) was used as the reference genome for transcriptome analysis. By this approach, we found that MT regulates the synthesis of NO and H2O2 and mediates the synthesis of secondary metabolites GRA and SF. GO annotations indicated that DEGs involved in the MT treatment of broccoli hairy roots were mainly related to catalytic activity, cells, and metabolic processes; the KEGG pathway analysis indicated that MT treatment likely affects the hormone signal transduction process in broccoli hairy roots; broccoli hairy roots were treated with 500 μmol/L MT for 0, 6, 12, 20, and 32 h, respectively; compared with 0 h, the yield of GRA and SF increased under the other treatments. The highest yields of GRA and SF occurred at 12 h. The NO content was the highest at 12 h, and the H2O2 content was positively correlated with MT concentration. The content of NO and H2O2 were regulated, and the content of GRA and SF was increased under MT treatment. NO synthase inhibitor (L-NAME and TUN) could effectively inhibit the content of NO in broccoli hairy roots and reduce GRA and SF yield; MT could regulate NO levels by regulating NO synthesis-related enzymes and could alleviate the reduction of NO content in tissue cells caused by NO synthase inhibitor and promote NO synthesis. These results have important theoretical implications for understanding the regulation of GRA and SF synthesis events by NO and H2O2. [ABSTRACT FROM AUTHOR]

Diets rich in cruciferous vegetables have been associated with a lower risk of incidence and progression of prostate cancer. Sulforaphane, an isothiocyanate derived from 4-methylsulphinylbutyl glucosinolate (glucoraphanin) that accumulates in certain of these vegetables, notably broccoli, has been implicated in their protective effects. Likewise, the consumption of garlic and its sulphur-containing compounds such as alliin have been associated with a reduction in risk of prostate cancer. In this study, we tested whether consuming glucoraphanin derived from broccoli seeds and alliin derived from garlic resulted in the occurrence of these potential bioactive compounds in the prostate, which may contribute to our understanding of the putative protective effects of these dietary components. We recruited 42 men scheduled for a trans-perineal prostate biopsy into a randomised, double-blinded, 2 × 2-factorial dietary supplement four-week intervention study, and 39 completed the study. The two active interventions were supplements providing glucoraphanin from broccoli (BroccoMax®) and alliin from garlic (Kwai Heartcare®). Following the intervention, prostate biopsy tissue was analysed for the presence of sulforaphane and its thiol conjugates and for alliin and associated metabolites. Sulforaphane occurred in significantly higher levels in the prostate tissue (both within the transition and peripheral zone) of men consuming the glucoraphanin containing supplements (p < 0.0001) compared to men not consuming these supplements. However, while alliin and alliin-derived metabolites were detected within the prostate, there was no significant difference in the concentrations of these compounds in the prostate of men consuming supplements derived from garlic compared to men not consuming these supplements. [ABSTRACT FROM AUTHOR]

Sulforaphane (SFN) is a potent activator of the transcriptional factor, Nuclear Factor Erythroid 2 (NF-E2)-Related factor 2 (NRF2). SFN and its precursor, glucoraphanin (sulforaphane glucosinolate, SGS), have been shown to ameliorate cognitive function in clinical trials and in vivo studies. However, the effects of SGS on age-related cognitive decline in Senescence-Accelerated Mouse Prone 8 (SAMP8) is unknown. In this study, we determined the preventive potential of SGS on age-related cognitive decline. One-month old SAMP8 mice or control SAM resistance 1 (SAMR1) mice were fed an ad libitum diet with or without SGS-containing broccoli sprout powder (0.3% w/w SGS in diet) until 13 months of age. SGS significantly improved long-term memory in SAMP8 at 12 months of age. Interestingly, SGS increased hippocampal mRNA and protein levels of peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC1α) and mitochondrial transcription factor A (TFAM), which are master regulators of mitochondrial biogenesis, both in SAMR1 and SAMP8 at 13 months of age. Furthermore, mRNAs for nuclear respiratory factor-1 (NRF-1) and mitochondrial DNA-encoded respiratory complex enzymes, but not mitochondrial DNA itself, were increased by SGS in SAMP8 mice. These results suggest that SGS prevents age-related cognitive decline by maintaining mitochondrial function in senescence-accelerated mice. [ABSTRACT FROM AUTHOR]

Intestinal permeabilization is central to the pathophysiology of chronic gut inflammation. This study investigated the efficacy of glucoraphanin (GR), prevalent in cruciferous vegetables, particularly broccoli, and its derivative sulforaphane (SF), in inhibiting tumor necrosis factor alpha (TNFα)-induced Caco-2 cell monolayers inflammation and permeabilization through the regulation of redox-sensitive events. TNFα binding to its receptor led to a rapid increase in oxidant production and subsequent elevation in the mRNA levels of NOX1, NOX4, and Duox2. GR and SF dose-dependently mitigated both these short- and long-term alterations in redox homeostasis. Downstream, GR and SF inhibited the activation of the redox-sensitive signaling cascades NF-κB (p65 and IKK) and MAPK ERK1/2, which contribute to inflammation and barrier permeabilization. GR (1 μM) and SF (0.5-1 μM) prevented TNFα-induced monolayer permeabilization and the associated reduction in the levels of the tight junction (TJ) proteins occludin and ZO-1. Both GR and SF also mitigated TNFα-induced increased mRNA levels of the myosin light chain kinase, which promotes TJ opening. Molecular docking suggests that although GR is mostly not absorbed, it could interact with extracellular and membrane sites in NOX1. Inhibition of NOX1 activity by GR would mitigate TNFα receptor downstream signaling and associated events. These findings support the concept that not only SF, but also GR, could exert systemic health benefits by protecting the intestinal barrier against inflammation-induced permeabilization, in part by regulating redox-sensitive pathways. GR has heretofore not been viewed as a biologically active molecule, but rather, the benign precursor of highly active SF. The consumption of GR and/or SF-rich vegetables or supplements in the diet may offer a means to mitigate the detrimental consequences of intestinal permeabilization, not only in disease states but also in conditions characterized by chronic inflammation of dietary and lifestyle origin., Competing Interests: Declaration of competing interest Authors have no conflict of interest to declare., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

The effects of a nanocarbon solution on the nutrients, glucosinolate metabolism and glucoraphanin pathway in broccoli were investigated. Significant positive linear relationships were observed between the nanocarbon solution and total protein yield, although effects on the soluble sugars, vitamin C and dry matter production were not observed. All nanocarbon solutions significantly increased the glucoraphanin content (p

Sarcopenia is a gradual and generalized skeletal muscle (SKM) syndrome, characterized by the impairment of muscle components and functionality. Hydrogen sulfide (H2S), endogenously formed within the body from the activity of cystathionine-γ-lyase (CSE), cystathionine- β-synthase (CBS), and mercaptopyruvate sulfurtransferase, is involved in SKM function. Here, in an in vitro model of sarcopenia based on damage induced by dexamethasone (DEX, 1 μM, 48 h treatment) in C2C12-derived myotubes, we investigated the protective potential of exogenous and endogenous sources of H2S, i.e., glucoraphanin (30 μM), L-cysteine (150 μM), and 3-mercaptopyruvate (150 μM). DEX impaired the H2S signalling in terms of a reduction in CBS and CSE expression and H2S biosynthesis. Glucoraphanin and 3-mercaptopyruvate but not L-cysteine prevented the apoptotic process induced by DEX. In parallel, the H2S-releasing molecules reduced the oxidative unbalance evoked by DEX, reducing catalase activity, O2− levels, and protein carbonylation. Glucoraphanin, 3-mercaptopyruvate, and L-cysteine avoided the changes in myotubes morphology and morphometrics after DEX treatment. In conclusion, in an in vitro model of sarcopenia, an impairment in CBS/CSE/H2S signalling occurs, whereas glucoraphanin, a natural H2S-releasing molecule, appears more effective for preventing the SKM damage. Therefore, glucoraphanin supplementation could be an innovative therapeutic approach in the management of sarcopenia. [ABSTRACT FROM AUTHOR]

Simple Summary: Cruciferous vegetable consumption is associated with reduced risk of tobacco-related cancers. Broccoli seed and sprout extract (BSSE) is rich in the isothiocyanate glucoraphanin, which is hydrolyzed to the phytochemical sulforaphane, a potent inducer of cytoprotective enzymes. This randomized, crossover trial evaluated detoxification of tobacco carcinogens by two doses of the BSSE Avmacol® in 49 otherwise healthy tobacco smokers. The higher dose (296 µmol of glucoraphanin daily) significantly upregulated detoxification of benzene, acrolein, and crotonaldehyde. Consumption of cruciferous vegetables, rich in the isothiocyanate glucoraphanin, is associated with reduced risk of tobacco-related cancers. Sulforaphane, released by hydrolysis of glucoraphanin, potently induces cytoprotective phase II enzymes. Sulforaphane decreased the incidence of oral cancer in the 4NQO carcinogenesis model. In residents of Qidong, China, broccoli seed and sprout extracts (BSSE) increased detoxification of air pollutants benzene and acrolein, also found in tobacco smoke. This randomized, crossover trial evaluated detoxification of tobacco carcinogens by the BSSE Avmacol® in otherwise healthy smokers. Participants were treated for 2 weeks with both low and higher-dose BSSE (148 µmol vs. 296 µmol of glucoraphanin daily), separated by a 2-week washout, with randomization to low-high vs. high-low sequence. The primary endpoint was detoxification of benzene, measured by urinary excretion of its mercapturic acid, SPMA. Secondary endpoints included bioavailability, detoxification of acrolein and crotonaldehyde, modulation by GST genotype, and toxicity. Forty-nine participants enrolled, including 26 (53%) females with median use of 20 cigarettes/day. Low and higher-dose BSSE showed a mean bioavailability of 11% and 10%, respectively. Higher-dose BSSE significantly upregulated urinary excretion of the mercapturic acids of benzene (p = 0.04), acrolein (p < 0.01), and crotonaldehyde (p = 0.02), independent of GST genotype. Retention and compliance were high resulting in early study completion. In conclusion, BSSE significantly upregulated detoxification of the tobacco carcinogens benzene, acrolein, and crotonaldehyde in current tobacco smokers. [ABSTRACT FROM AUTHOR]

The aim of this study was to compare the effects of thermosonication (18 kHz at 60 °C for 7 min) pre-treatment with thermal treatment alone (60 °C for 7 min) of broccoli florets prior to pureeing and fermentation on selected bioactive components of fermented broccoli puree. Both thermal and thermosoncation pre-treatments significantly increased the rate of acidification of broccoli puree compared to control untreated broccoli puree, with the time to reach pH 4 being 8.25, 9.9, and 24 h, respectively, for thermally treated, thermosonicated, and control samples. The highest sulforaphane yield of 7268 µmol/kg dry weight (DW) was observed in the thermosonicated samples, followed by 6227 µmol/kg DW and 3180 µmol/kg DW in the thermally treated and untreated samples, respectively. The measurable residual glucoraphanin content was 1642 µmol/kg DW, 1187 µmol/kg DW, and 1047 µmol/kg DW, respectively, in the thermonsonicated, thermally pre-treated, and control fermented samples, indicating that pre-treatment specially by thermosonication increases the extractability of glucoraphanin. The higher sulforaphane yield in the thermosonicated and thermally pre-treated samples could be due to increased extractability and accessibility of glucoraphanin and interaction with myrosinase in addition to the inactivation of epthiospecifier protein (ESP), which directs conversion away from sulforaphane into sulforaphane nitrile. [ABSTRACT FROM AUTHOR]

Broccoli cultivars that have enhanced accumulation of methionine-derived glucosinolates have been developed through the introgression of a novel allele of the MYB28 transcription factor from the wild species Brassica villosa. Through a novel k-mer approach, we characterised the extent of the introgression of unique B. villosa genome sequences into high glucosinolate broccoli genotypes. RNAseq analyses indicated that the introgression of the B. villosa MYB28 C2 allele resulted in the enhanced expression of the MYB28 transcription factor, and modified expression of genes associated with sulphate absorption and reduction, and methionine and glucosinolate biosynthesis when compared to standard broccoli. A adenine-thymine (AT) short tandem repeat (STR) was identified within the 5′ untranslated region (UTR) B. villosa MYB28 allele that was absent from two divergent cultivated forms of Brassica oleracea , which may underpin the enhanced expression of B. villosa MYB28. [ABSTRACT FROM AUTHOR]

Broccoli, a Brassica crop, is an important vegetative crop because of its nutritional components and beneficial phytochemicals. Glucoraphanin (GR), a major glucosinolate (GSL) in broccoli, is converted by hydrolyzation of the endogenous enzyme, myrosinase, into sulforaphane (SR), which protects the body against a variety of chronic diseases. Despite their economic importance, biotechnological approaches for increasing GR content in Brassica species are still limited. The main objective of this study was to develop a GR-rich broccoli cultivar using the CRISPR/Cas9-mediated DNA-free genome-editing technique. It is considered that MYB28 is one of the key genes involved in the accumulation of GSL levels in broccoli. Furthermore, with increased GSL levels by introgression of MYB28 from wild species, B. villosa showed a 9 bp deletion in exon 3, leading to one amino acid substitution and the deletion of three amino acids. Therefore, we considered the 9 bp deletion to be the most significant change in GR-rich broccoli and conducted Cas9 protein and single-guide RNA transfection into broccoli protoplasts for editing the flanking sequence of the 9 bp deleted MYB28 gene. Finally, increased GR content was observed in broccoli regenerated from protoplasts with specifically edited MYB28. [ABSTRACT FROM AUTHOR]

Broccoli cultivars that have enhanced accumulation of methionine-derived glucosinolates have been developed through the introgression of a novel allele of the MYB28 transcription factor from the wild species Brassica villosa. Through a novel k-mer approach, we characterised the extent of the introgression of unique B. villosa genome sequences into high glucosinolate broccoli genotypes. RNAseq analyses indicated that the introgression of the B. villosa MYB28 C2 allele resulted in the enhanced expression of the MYB28 transcription factor, and modified expression of genes associated with sulphate absorption and reduction, and methionine and glucosinolate biosynthesis when compared to standard broccoli. A adenine-thymine (AT) short tandem repeat (STR) was identified within the 5′ untranslated region (UTR) B. villosa MYB28 allele that was absent from two divergent cultivated forms of Brassica oleracea, which may underpin the enhanced expression of B. villosa MYB28.

In this investigation, the combinations of exogenous pretreatment (melatonin or vitamin C) and contact ultrasound-assisted air drying were utilized to dry broccoli florets. To understand the influences of the studied dehydration methods on the conversion of glucoraphanin to bioactive sulforaphane in broccoli, various components (like glucoraphanin, sulforaphane, myrosinase, etc.) and factors (temperature and moisture) involved in the metabolism pathway were analyzed. The results showed that compared with direct air drying, the sequential exogenous pretreatment and contact ultrasound drying shortened the drying time by 19.0–22.7%. Meanwhile, contact sonication could promote the degradation of glucoraphanin. Both melatonin pretreatment and vitamin C pretreatment showed protective effects on the sulforaphane content and myrosinase activity during the subsequent drying process. At the end of drying, the sulforaphane content in samples dehydrated by the sequential melatonin (or vitamin C) pretreatment and ultrasound-intensified drying was 14.4% (or 26.5%) higher than only air-dried samples. The correlation analysis revealed that the exogenous pretreatment or ultrasound could affect the enzymatic degradation of glucoraphanin and the generation of sulforaphane through weakening the connections of sulforaphane-myrosinase, sulforaphane-VC, and VC-myrosinase. Overall, the reported results can enrich the biochemistry knowledge about the transformation of glucoraphanin to sulforaphane in cruciferous vegetables during drying, and the combined VC/melatonin pretreatment and ultrasound drying is conducive to protect bioactive sulforaphane in dehydrated broccoli.

Sulforaphane (SF) is one of the most effective natural products in preventing and fighting cancer, found in cruciferous plants. In this study, broccoli hairy roots grown for 20 d were used as the experimental material, and it was treated with 500 μmol/L melatonin (MT) for 0, 12 and 32 h to explore the effect of MT on the conversion of glucoraphanin (GRA) to SF. Results showed that the yields of GRA and SF were the largest under MT treatment for 12 h, which were 1.53 and 1.93-fold, respectively, compared to 0 h. However, Myrosinases activity was the highest under MT treatment for 32 h, which was 1.42-fold compared to that of the 0 h. The differential expression of key genes involved in GRA conversion to SF in broccoli hairy roots was identified transcriptome sequencing, and the path of the transformation from GRA to SF was simulated, which provided a theoretical basis for establishing an efficient transformation system from GRA to SF. [ABSTRACT FROM AUTHOR]

Abstract Aim Epidemiological data suggest that maternal immune activation (MIA) plays a role in the etiology of neuropsychiatric disorders including autism spectrum disorder (ASD) and schizophrenia. However, there is no prophylactic nutrition that can prevent the onset of neurodevelopmental disorders in offspring after MIA. The aim of this study was undertaken to examine whether dietary intake of glucoraphanin (GF: the precursor of a natural anti‐inflammatory compound sulforaphane) can prevent the onset of behavioral abnormalities in offspring after MIA. Methods One percent of GF food pellet or normal food pellet was given into female mice during pregnancy and lactation (from E5 to P21). Saline (5 mL/kg/d) or poly(I:C) (5 mg/kg/d) was injected into pregnant mice from E12 to E17. Behavioral tests and immunohistochemistry of parvalbumin (PV) were performed in male offspring. Results Dietary intake of GF during pregnancy and lactation prevented cognitive deficits and social interaction deficits in the juvenile offspring after MIA. Furthermore, dietary intake of GF during pregnancy and lactation prevented cognitive deficits in the adult offspring after MIA. Moreover, dietary intake of GF prevented the reduction of PV immunoreactivity in the medial prefrontal cortex of adult offspring after MIA. Conclusion These data suggest that dietary intake of GF during pregnancy and lactation could prevent behavioral abnormalities in offspring after MIA.

Abstract Sulforaphane (SFR), an anticarcinogenic compound, forms from the hydrolysis of glucoraphanin (GLR) in broccoli. Cooking methods have been shown to affect broccoli GLR and SFR levels, but little is known about the effect of lightly cooking processes on them. In this study, the effects of microwave and low‐temperature cooking on GLR and SFR contents in broccoli were investigated. Both microwaving and mild heating increased the GLR and SFR levels in broccoli compared to the raw samples (without any treatment). In particular, SFR level was significantly low under 40°C and dramatically increased from 40 to 60°C, but nothing was detected at 70°C. Compared with conventional heating, microwave heating increased the GLR and SFR yield by about 80% at 50 and 60°C. Microwave power level also influenced the SFR contents. At the same temperatures (50 and 60°C), high‐power microwave (950 W) with a short time produced over 40% more SFR than low‐power microwave treatment (475 W). Hence, mild heating by microwave could increase the GLR and SFR levels in broccoli, and high‐power microwave heating with temperature control at 60°C could retain higher bioavailability of these bioactive compounds in broccoli.

As the prevalence of inflammatory bowel diseases (IBD) rises, the etiology of IBD draws increasing attention. Glucoraphanin (GRP), enriched in cruciferous vegetables, is a precursor of sulforaphane, known to have anti-inflammatory and antioxidative effects. We hypothesized that dietary GRP supplementation can prevent mitochondrial dysfunction and oxidative stress in an acute colitis mouse model induced by dextran sulfate sodium (DSS). Eight-week-old mice were fed a regular rodent diet either supplemented with or without GRP. After 4 weeks of dietary treatments, half of the mice within each dietary group were subjected to 2.5% DSS treatment to induce colitis. Dietary GRP decreased DSS-induced body weight loss, disease activity index, and colon shortening. Glucoraphanin supplementation protected the colonic histological structure, suppressed inflammatory cytokines, interleukin (IL)-1β, IL-18, and tumor necrosis factor-α (TNF-α), and reduced macrophage infiltration in colonic tissues. Consistently, dietary GRP activated AMP-activated protein kinase (AMPK), peroxisome proliferator-activated receptor-gamma coactivator (PGC)-1α, and nuclear factor erythroid 2-related factor 2 (NRF2) pathways in the colonic tissues of DSS-treated mice, which was associated with increased mitochondrial DNA and decreased content of the oxidative product 8-hydroxydeoxyguanosine (8-OHDG), a nucleotide oxidative product of DNA. In conclusion, dietary GRP attenuated mitochondrial dysfunction, inflammatory response, and oxidative stress induced by DSS, suggesting that dietary GRP provides a dietary strategy to alleviate IBD symptoms.

Myrosinase (Myr) catalyzes the hydrolysis of glucosinolates, yielding biologically active metabolites. In this study, glucoraphanin (GRA) extracted from broccoli seeds was effectively hydrolyzed using a Myr-obtained cabbage aphid ( Brevicoryne brassicae ) ( Bb Myr) to produce (R)-sulforaphane (SFN). The gene encoding Bb Myr was successfully heterologously expressed in Escherichia coli , resulting in the production of 1.6 g/L (R)-SFN, with a remarkable yield of 20.8 mg/g broccoli seeds , achieved using recombination E. coli whole-cell catalysis under optimal conditions (pH 4.5, 45 °C). Subsequently, Bb Myr underwent combinatorial simulation-driven mutagenesis, yielding a mutant, DE9 (N321D/Y426S), showing a remarkable 2.91-fold increase in the catalytic efficiency ( k cat / K M ) compared with the original enzyme. Molecular dynamics simulations demonstrated that the N321D mutation in loopA of mutant DE9 enhanced loopA stability by inducing favorable alterations in hydrogen bonds, while the Y426S mutation in loopB decreased spatial resistance. This research lays a foundation for the environmentally sustainable enzymatic (R)-SFN synthesis.

Scope: Obesity and its metabolic comorbidities pose a major global challenge for public health. Glucoraphanin (GRN) is a natural bioactive compound enriched in broccoli that is known to have potential health benefits against various human chronic diseases., Methods and Results: This study investigats the effects of broccoli GRN supplementation on body weight, metabolic parameters, gut microbiome and metabolome associated with obesity. The study is conducted on an obese-related C57BL/6J mouse model through the treatment of normal control diet, high-fat diet (HFD)and GRN-supplemented HFD (HFD-GRN) to determine the metabolic protection of GRN. The results shows that GRN treatment alleviates obesity-related traits leading to improved glucose metabolism in HFD-fed animals. Mechanically, the study noticed that GRN significantly shifts the gut microbial diversity and composition to an eubiosis status. GRN supplement also significantly alters plasma metabolite profiles. Further integrated analysis reveal a complex interaction between the gut microbes and host metabolism that may contribute to GRN-induced beneficial effects against HFD., Conclusion: These results indicate that beneficial effects of broccoli GRN on reversing HFD-induced adverse metabolic parameters may be attributed to its impacts on reprogramming microbial community and metabolites. Identification of the mechanistic functions of GRN further warrants it as a dietary candidate for obesity prevention., (© 2024 The Authors. Molecular Nutrition & Food Research published by Wiley‐VCH GmbH.)