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Background: Asthma affects 3% of the global population, leading to over 0.25 million deaths. Due to its complexity, asthma is difficult to cure or prevent, and current therapies have limitations. This has led to a growing demand for alternative asthma treatments. We found rosmarinic acid (RosA) as a potential new drug candidate from natural medicine. However, RosA has poor bioavailability and remains mainly in the gastrointestinal tract after oral administration, suggesting the involvement of gut microbiota in its bioactivity. Purpose: To investigate the mechanism of RosA in alleviating allergic asthma by gut-lung axis. Methods: We used 16S rRNA gene sequencing and metabolites analysis to investigate RosA's modulation of gut microbiota. Techniques of molecular biology and metabolomics were employed to study the pharmacological mechanism of RosA. Cohousing was used to confirm the involvement of gut microbiota in RosA-induced improvement of allergic asthma. Results: RosA decreased cholate levels from spore-forming bacteria, leading to reduced 5-hydroxytryptamine (5HT) synthesis, bronchoconstriction, vasodilation, and inflammatory cell infiltration. It also increased short-chain fatty acids (SCFAs) levels, facilitating the expression of intestinal tight junction proteins to promote intestinal integrity. SCFAs upregulated intestinal monocarboxylate transporters (MCTs), thereby improving their systemic delivery to reduce Th2/ILC2 mediated inflammatory response and suppress eosinophil influx and mucus production in lung. Additionally, RosA inhibited lipopolysaccharide (LPS) production and translocation, leading to reduced TLR4-NF kappa B mediated pulmonary inflammation and oxidative stress. Conclusions: The anti-asthmatic mechanism of oral RosA is primarily driven by modulation of gut microbiotaderived 5-HT, SCFAs, and LPS, achieving a combined synergistic effect. RosA is a safe, effective, and reliable drug candidate that could potentially replace glucocorticoids for ast

The aim of this study was to comparatively analyze chemical composition and biological activity of wild- and in vitro grown Rindera umbellata. Explants were cultivated on 0.003–0.3 M sucrose, fructose, or glucose. HPLC-DAD for quantifying rosmarinic (RA) and lithospermic B (LAB) acids and GC-MS/FID for qualitative pyrrolizidine alkaloids (PAs) detection were used. Antioxidant activity (DPPH and ABTS assays) and cytotoxicity (MTT test) were monitored. Identified PAs were 7-angeloyl heliotridane, lindelofine, 7-angeloyl heliotridine, 7-angeloyl-9-(+)-trachelanthylheliotridine, punctanecine, and heliosupine, with higher variability reported in wild-growing samples. Total phenolic contents (TPCs) were comparable in wild-growing and in vitro samples, but total flavonoid (TFC) and RA levels were multifold higher in in vitro samples. Notably, high concentration of LAB was detected in wild-growing roots. Amounts of 0.3 M and 0.1 M of sucrose were optimal for TFC and RA production, while maximal antioxidant activity was monitored in plants grown on 0.3 M sucrose. The MTT test indicated colorectal HT-29 as more sensitive than A549 lung adenocarcinoma and normal MRC-5 cells, showing selective sensitivity to wild-growing and 0.3 M sucrose samples. In conclusion, PAs in vitro, as well as TPC, TFC, RA, and LAB in both growing conditions were detected for the first time in R. umbellata.

Comfrey (Symphytum officinale L.) roots are well-known bioactive ingredients included in various cosmeceutical and pharmaceutical preparations. In this study, the influence of the post-harvest storage on the chemico-biological potential of roots collected from different European regions and stored for up to six months was investigated. Total phenolic content (TPC) and total phenolic acid content (TPAC) were spectrophotometrically estimated, whereas the levels of individual phenolic and pyrrolizidine alkaloidal markers were determined by HPLC-DAD and HPLC-MS/MS, respectively. The changes in the biological potential was tracked via antioxidant (DPPH, ABTS, CUPRAC, and FRAP) and anti-enzymatic (cholinesterase, tyrosinase, glucosidase, and amylase) assays. TPC and TPAC varied from 6.48-16.57 mg GAE/g d.w. root and from 2.67-9.03 mg CAE/g, respectively. The concentration of the four phenolics (rosmarinic acid, globoidnan A, globoidnan B, rabdosiin) and six pyrrolizidine alkaloids generally showed maximum values at 1-3 months, after which their levels significantly decreased. With respect to the bioassays, the samples showed a wide range of antioxidant and anti-enzymatic effects; however, a direct storage time-bioactivity relationship was not observed. Similar conclusions were also revealed by the multivariate and correlation analyses. Our study could improve the current knowledge of the shelf-life properties of comfrey-based products and enhance their industrial exploitation.

The growing interest in rosmarinic acid (RA), an ester of caffeic acid and 3,4‐dihydroxyphenyl lactic acid, is due to its biological activities, which include cognitive‐enhancing effects, slowing the development of Alzheimer's disease, cancer chemoprotection, and anti‐inflammatory activity. Inspired by the challenge of meeting the growing demand for this plant secondary metabolite, we developed a biotechnological platform based on cell suspension cultures of Satureja khuzistanica. The high amounts of RA produced by this system accumulated mainly inside the cells. To further improve production, two elicitors, 100 μM methyl jasmonate (MeJA) and 40 mM cyclodextrin (CD), were tested, separately and together. MeJA increased RA productivity more than 3‐fold, the elicited cultures achieving an RA production of 3.9 g L−1 without affecting biomass productivity. CD did not have a clear effect on RA production, and under the combined treatment of MeJA + CD only a small amount of RA was released to the medium. When the cell culture was transferred from a shake flask to a wave‐mixed bioreactor, a maximum RA production of 3.1 g L−1 and biomass productivity of 18.7 g L−1 d−1 was achieved under MeJA elicitation, demonstrating the suitability of S. khuzistanica cell suspensions for the biotechnological production of this bioactive plant secondary metabolite.

Ulcerative colitis (UC), one of the two main types of inflammatory bowel disease, has no effective treatment. Rosmarinic acid (RA) is a polyphenol that, when administered orally, is metabolised in the small intestine, compromising its beneficial effects. We used chitosan/nutriosecoated niosomes loaded with RA to protect RA from gastric degradation and target the colon and evaluated their effect on acute colitis induced by 4% dextran sodium sulphate (DSS) for seven days in mice. RA-loaded nanovesicles (5, 10 and 20 mg/kg) or free RA (20 mg/kg) were orally administered from three days prior to colitis induction and during days 1, 3, 5 and 7 of DSS administration. RA-loaded nanovesicles improved body weight loss and disease activity index as well as increased mucus production and decreased myeloperoxidase activity and TNF-_ production. Moreover, RA-loaded nanovesicles downregulated protein expression of inflammasome components such as NLR family pyrin domain-containing 3 (NLRP3), adaptor protein (ASC) and caspase-1, and the consequent reduction of IL-1_ levels. Furthermore, nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) protein expression increased after the RA-loaded nanovesicles treatment However, these mechanistic changes were not detected with the RA-free treatment. Our findings suggest that the use of chitosan/nutriose-coated niosomes to increase RA local bioavailability could be a promising nutraceutical strategy for oral colon-targeted UC therapy.

Ulcerative colitis (UC), one of the two main types of inflammatory bowel disease, has no effective treatment. Rosmarinic acid (RA) is a polyphenol that, when administered orally, is metabolised in the small intestine, compromising its beneficial effects. We used chitosan/nutriosecoated niosomes loaded with RA to protect RA from gastric degradation and target the colon and evaluated their effect on acute colitis induced by 4% dextran sodium sulphate (DSS) for seven days in mice. RA-loaded nanovesicles (5, 10 and 20 mg/kg) or free RA (20 mg/kg) were orally administered from three days prior to colitis induction and during days 1, 3, 5 and 7 of DSS administration. RA-loaded nanovesicles improved body weight loss and disease activity index as well as increased mucus production and decreased myeloperoxidase activity and TNF-_ production. Moreover, RA-loaded nanovesicles downregulated protein expression of inflammasome components such as NLR family pyrin domain-containing 3 (NLRP3), adaptor protein (ASC) and caspase-1, and the consequent reduction of IL-1_ levels. Furthermore, nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) protein expression increased after the RA-loaded nanovesicles treatment However, these mechanistic changes were not detected with the RA-free treatment. Our findings suggest that the use of chitosan/nutriose-coated niosomes to increase RA local bioavailability could be a promising nutraceutical strategy for oral colon-targeted UC therapy.

Modern lifestyle factors, such as physical inactivity, obesity, smoking, and exposure to environmental pollution, induce excessive generation of free radicals and reactive oxygen species (ROS) in the body. These by-products of oxygen metabolism play a key role in the development of various human diseases such as cancer, diabetes, heart failure, brain damage, muscle problems, premature aging, eye injuries, and a weakened immune system. Synthetic and natural antioxidants, which act as free radical scavengers, are widely used in the food and beverage industries. The toxicity and carcinogenic effects of some synthetic antioxidants have generated interest in natural alternatives, especially plant-derived polyphenols (e.g., phenolic acids, flavonoids, stilbenes, tannins, coumarins, lignins, lignans, quinines, curcuminoids, chalcones, and essential oil terpenoids). This review focuses on the well-known phenolic antioxidant rosmarinic acid (RA), an ester of caffeic acid and (R)-(+)-3-(3,4-dihydroxyphenyl) lactic acid, describing its wide distribution in thirty-nine plant families and the potential productivity of plant sources. A botanical and phytochemical description is provided of a new rich source of RA, Satureja khuzistanica Jamzad (Lamiaceae). Recently reported approaches to the biotechnological production of RA are summarized, highlighting the establishment of cell suspension cultures of S. khuzistanica as an RA chemical biofactory.

Ulcerative colitis (UC), one of the two main types of inflammatory bowel disease, has no effective treatment. Rosmarinic acid (RA) is a polyphenol that, when administered orally, is metabolised in the small intestine, compromising its beneficial effects. We used chitosan/nutriosecoated niosomes loaded with RA to protect RA from gastric degradation and target the colon and evaluated their effect on acute colitis induced by 4% dextran sodium sulphate (DSS) for seven days in mice. RA-loaded nanovesicles (5, 10 and 20 mg/kg) or free RA (20 mg/kg) were orally administered from three days prior to colitis induction and during days 1, 3, 5 and 7 of DSS administration. RA-loaded nanovesicles improved body weight loss and disease activity index as well as increased mucus production and decreased myeloperoxidase activity and TNF-_ production. Moreover, RA-loaded nanovesicles downregulated protein expression of inflammasome components such as NLR family pyrin domain-containing 3 (NLRP3), adaptor protein (ASC) and caspase-1, and the consequent reduction of IL-1_ levels. Furthermore, nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) protein expression increased after the RA-loaded nanovesicles treatment However, these mechanistic changes were not detected with the RA-free treatment. Our findings suggest that the use of chitosan/nutriose-coated niosomes to increase RA local bioavailability could be a promising nutraceutical strategy for oral colon-targeted UC therapy.

Serum amyloid A (SAA) is one of the most important precursor amyloid proteins and plays a vital step in AA amyloidosis, although the underlying aggregation mechanism has not been elucidated. Since SAA aggregation is a key step in this pathogenesis, inhibitors are useful to prevent and treat AA amyloidosis, serving as tools to investigate the pathogenic mechanism. In this study, we showed that rosmarinic acid (RA), which is a well-known inhibitor of the aggregation of amyloid beta (A beta), displayed inhibitory activity against SAA aggregation in vitro using a microliter-scale high-throughput screening (MSHTS) system with quantum-dot nanoprobes. Therefore, we evaluated the amyloid aggregation inhibitory activity of blood and the deposition of SAA in organs by feeding mice withMelissa officinalisextract (ME) containing RA as an active substance. Interestingly, the inhibitory activity of ME-fed mice sera for SAA and A beta aggregation, measured with the MSHTS system, was higher than that of the control group. The amount of amyloid deposition in the organs of ME-fed mice was lower than that in the control group, suggesting that the SAA aggregation inhibitory activity of serum is associated with SAA deposition. These results suggest that dietary intake of RA-containing ME enhanced amyloid aggregation inhibitory activity of blood and suppressed SAA deposition in organs. This study also demonstrated that the MSHTS system could be applied to in vitro screening and to monitor comprehensive activity of metabolized foods adsorbed by blood.

Modern lifestyle factors, such as physical inactivity, obesity, smoking, and exposure to environmental pollution, induce excessive generation of free radicals and reactive oxygen species (ROS) in the body. These by-products of oxygen metabolism play a key role in the development of various human diseases such as cancer, diabetes, heart failure, brain damage, muscle problems, premature aging, eye injuries, and a weakened immune system. Synthetic and natural antioxidants, which act as free radical scavengers, are widely used in the food and beverage industries. The toxicity and carcinogenic effects of some synthetic antioxidants have generated interest in natural alternatives, especially plant-derived polyphenols (e.g., phenolic acids, flavonoids, stilbenes, tannins, coumarins, lignins, lignans, quinines, curcuminoids, chalcones, and essential oil terpenoids). This review focuses on the well-known phenolic antioxidant rosmarinic acid (RA), an ester of caffeic acid and (R)-(+)-3-(3,4-dihydroxyphenyl) lactic acid, describing its wide distribution in thirty-nine plant families and the potential productivity of plant sources. A botanical and phytochemical description is provided of a new rich source of RA, Satureja khuzistanica Jamzad (Lamiaceae). Recently reported approaches to the biotechnological production of RA are summarized, highlighting the establishment of cell suspension cultures of S. khuzistanica as an RA chemical biofactory.

Ulcerative colitis (UC), one of the two main types of inflammatory bowel disease, has no effective treatment. Rosmarinic acid (RA) is a polyphenol that, when administered orally, is metabolised in the small intestine, compromising its beneficial effects. We used chitosan/nutriosecoated niosomes loaded with RA to protect RA from gastric degradation and target the colon and evaluated their effect on acute colitis induced by 4% dextran sodium sulphate (DSS) for seven days in mice. RA-loaded nanovesicles (5, 10 and 20 mg/kg) or free RA (20 mg/kg) were orally administered from three days prior to colitis induction and during days 1, 3, 5 and 7 of DSS administration. RA-loaded nanovesicles improved body weight loss and disease activity index as well as increased mucus production and decreased myeloperoxidase activity and TNF-_ production. Moreover, RA-loaded nanovesicles downregulated protein expression of inflammasome components such as NLR family pyrin domain-containing 3 (NLRP3), adaptor protein (ASC) and caspase-1, and the consequent reduction of IL-1_ levels. Furthermore, nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) protein expression increased after the RA-loaded nanovesicles treatment However, these mechanistic changes were not detected with the RA-free treatment. Our findings suggest that the use of chitosan/nutriose-coated niosomes to increase RA local bioavailability could be a promising nutraceutical strategy for oral colon-targeted UC therapy.

Rosmarinic acid is an attractive candidate for skin applications because of its antioxidant, anti-inflammatory, and photoprotective functions, however, its poor bioavailability hampers its therapeutic outcome. In this context, synthesis of polymer conjugates is an alternative to enlarge its applications. This work describes the synthesis of novel water-soluble chitosan – rosmarinic acid conjugates (CSRA) that have great potential for skin applications. Chitosan was functionalized with different contents of rosmarinic acid as confirmed by ATR-FTIR, H NMR and UV spectroscopies. CSRA conjugates presented three-fold radical scavenger capacity compared to the free phenolic compound. Films were prepared by solvent-casting procedure and the biological activity of the lixiviates was studied in vitro. Results revealed that lixiviates reduced activation of inflamed macrophages, improved antibacterial capacity against E. coli with respect to native chitosan and free rosmarinic acid, and also attenuated UVB-induced cellular damage and reactive oxygen species production in fibroblasts and keratinocytes.

Introduction: Symphytum officinale L. (comfrey, Boraginaceae) is a cultivated or spontaneously growing medicinal plant that is traditionally used for the treatment of bone fractures, hematomas, muscle pains and joint pains. A wide range of topical preparations and dried roots for ex tempore applications are marketed in European drug stores or pharmacies. Objective: The aim of this study was to perform the qualitative and quantitative analysis of pyrrolizidine alkaloids (PAs) and phenolic compounds in the hydroethanolic extracts of 16 commercial comfrey root batches purchased from 12 different European countries. Methods: Liquid chromatography hyphenated with high-resolution tandem mass spectrometry (LC-HRMS/MS) was used for the profiling of PAs and phenolic compounds, whereas LC-MS/MS and liquid chromatography with diode array detection (LC-DAD) were used for their quantification. Results: 20 PAs (i.e. intermedine, lycopsamine, acetylintermedine, acetyllycopsamine, symphytine, symphytine-N-oxide), 17 phenolic compounds (i.e. caffeic and rosmarinic acids, rabdosiin, globoidnan A, globoidnan B) and 9 nonphenolic compounds (sugars, organic and fatty acids) were fully or partly annotated in the analysed samples. In addition, the quantitative analyses revealed that globoidnan B, rabdosiin and globoidnan A are new phenolic markers that can be used together with rosmarinic acid and PAs for the quality control of commercial comfrey root batches. Conclusions: This study brings new insights into the phytochemical complexity of S. officinale, revealing not only numerous toxic PAs, but also a significant number of valuable phenolic compounds that could contribute to the bioactivities of comfreybased preparations

Many medicinal plant species are currently threatened in their natural habitats because of the growing demand for phytochemicals worldwide. A sustainable alternative for the production of bioactive plant compounds are plant biofactories based on cell cultures and organs. In addition, plant extracts from biofactories have significant advantages over those obtained from plants, since they are free of contamination by microorganisms, herbicides and pesticides, and they provide more stable levels of active ingredients. In this context, we report the establishment of Satureja khuzistanica cell cultures able to produce high amounts of rosmarinic acid (RA). The production of this phytopharmaceutical was increased when the cultures were elicited with coronatine and scaled up to a benchtop bioreactor. S. khuzistanica extracts enriched in RA were found to reduce the viability of cancer cell lines, increasing the sub-G0/G1 cell population and the activity of caspase-8 in MCF-7 cells, which suggest that S. khuzistanica extracts can induce apoptosis of MCF-7 cells through activation of the extrinsic pathway. In addition, our findings indicate that other compounds in S. khuzistanica extracts may act synergistically to potentiate the anticancer activity of RA.

Serum amyloid A (SAA) is one of the most important precursor amyloid proteins and plays a vital step in AA amyloidosis, although the underlying aggregation mechanism has not been elucidated. Since SAA aggregation is a key step in this pathogenesis, inhibitors are useful to prevent and treat AA amyloidosis, serving as tools to investigate the pathogenic mechanism. In this study, we showed that rosmarinic acid (RA), which is a well-known inhibitor of the aggregation of amyloid beta (A beta), displayed inhibitory activity against SAA aggregation in vitro using a microliter-scale high-throughput screening (MSHTS) system with quantum-dot nanoprobes. Therefore, we evaluated the amyloid aggregation inhibitory activity of blood and the deposition of SAA in organs by feeding mice withMelissa officinalisextract (ME) containing RA as an active substance. Interestingly, the inhibitory activity of ME-fed mice sera for SAA and A beta aggregation, measured with the MSHTS system, was higher than that of the control group. The amount of amyloid deposition in the organs of ME-fed mice was lower than that in the control group, suggesting that the SAA aggregation inhibitory activity of serum is associated with SAA deposition. These results suggest that dietary intake of RA-containing ME enhanced amyloid aggregation inhibitory activity of blood and suppressed SAA deposition in organs. This study also demonstrated that the MSHTS system could be applied to in vitro screening and to monitor comprehensive activity of metabolized foods adsorbed by blood.

Serum amyloid A (SAA) is one of the most important precursor amyloid proteins and plays a vital step in AA amyloidosis, although the underlying aggregation mechanism has not been elucidated. Since SAA aggregation is a key step in this pathogenesis, inhibitors are useful to prevent and treat AA amyloidosis, serving as tools to investigate the pathogenic mechanism. In this study, we showed that rosmarinic acid (RA), which is a well-known inhibitor of the aggregation of amyloid beta (A beta), displayed inhibitory activity against SAA aggregation in vitro using a microliter-scale high-throughput screening (MSHTS) system with quantum-dot nanoprobes. Therefore, we evaluated the amyloid aggregation inhibitory activity of blood and the deposition of SAA in organs by feeding mice withMelissa officinalisextract (ME) containing RA as an active substance. Interestingly, the inhibitory activity of ME-fed mice sera for SAA and A beta aggregation, measured with the MSHTS system, was higher than that of the control group. The amount of amyloid deposition in the organs of ME-fed mice was lower than that in the control group, suggesting that the SAA aggregation inhibitory activity of serum is associated with SAA deposition. These results suggest that dietary intake of RA-containing ME enhanced amyloid aggregation inhibitory activity of blood and suppressed SAA deposition in organs. This study also demonstrated that the MSHTS system could be applied to in vitro screening and to monitor comprehensive activity of metabolized foods adsorbed by blood.

Alzheimer’s disease is the most common neurodegenerative disease and it is believed that around 46.8 million of persons suffer this type of dementia. Nowadays, the disease has an increased prevalence and incidence, however current drugs only reduce symptoms and try to extend the life of the patient. Many of the presently treatments are incapable of crossing the blood-brain barrier (BBB), for this reason we have designed a drug delivery system based on poly-(lactic-co-gycolic acid) (PLGA) that has been reported to have advantageous properties like biocompatibility and biodegradability that ensures safe therapies. In this work, we develop a formulation based on a polymeric matrix that contains Rosmarinic acid (RA), which is a therapeutic agent against Alzheimer’s disease. The nanoparticles were prepared by nano-emulsion templating using a low-energy method, the Phase inversion composition (PIC) method. This proposal is a simple and very versatile technology, which allows particle size control and can be performed at mild conditions. PLGA-Rosmarinic acid- loaded nanoparticles were characterized and the polymeric surface was functionalized. There is still a long way to go in this field in order to find the root of the disease and thus be able to understand it better to elaborate a more effective treatment. This research is aimed at developing a formulation based on RA with the hope of bringing light to this field and expectations of continuance, ending in clinical trials in the future.

Many medicinal plant species are currently threatened in their natural habitats because of the growing demand for phytochemicals worldwide. A sustainable alternative for the production of bioactive plant compounds are plant biofactories based on cell cultures and organs. In addition, plant extracts from biofactories have significant advantages over those obtained from plants, since they are free of contamination by microorganisms, herbicides and pesticides, and they provide more stable levels of active ingredients. In this context, we report the establishment of Satureja khuzistanica cell cultures able to produce high amounts of rosmarinic acid (RA). The production of this phytopharmaceutical was increased when the cultures were elicited with coronatine and scaled up to a benchtop bioreactor. S. khuzistanica extracts enriched in RA were found to reduce the viability of cancer cell lines, increasing the sub-G0/G1 cell population and the activity of caspase-8 in MCF-7 cells, which suggest that S. khuzistanica extracts can induce apoptosis of MCF-7 cells through activation of the extrinsic pathway. In addition, our findings indicate that other compounds in S. khuzistanica extracts may act synergistically to potentiate the anticancer activity of RA.

Introduction: Symphytum officinale L. (comfrey, Boraginaceae) is a cultivated or spontaneously growing medicinal plant that is traditionally used for the treatment of bone fractures, hematomas, muscle pains and joint pains. A wide range of topical preparations and dried roots for ex tempore applications are marketed in European drug stores or pharmacies. Objective: The aim of this study was to perform the qualitative and quantitative analysis of pyrrolizidine alkaloids (PAs) and phenolic compounds in the hydroethanolic extracts of 16 commercial comfrey root batches purchased from 12 different European countries. Methods: Liquid chromatography hyphenated with high-resolution tandem mass spectrometry (LC-HRMS/MS) was used for the profiling of PAs and phenolic compounds, whereas LC-MS/MS and liquid chromatography with diode array detection (LC-DAD) were used for their quantification. Results: 20 PAs (i.e. intermedine, lycopsamine, acetylintermedine, acetyllycopsamine, symphytine, symphytine-N-oxide), 17 phenolic compounds (i.e. caffeic and rosmarinic acids, rabdosiin, globoidnan A, globoidnan B) and 9 nonphenolic compounds (sugars, organic and fatty acids) were fully or partly annotated in the analysed samples. In addition, the quantitative analyses revealed that globoidnan B, rabdosiin and globoidnan A are new phenolic markers that can be used together with rosmarinic acid and PAs for the quality control of commercial comfrey root batches. Conclusions: This study brings new insights into the phytochemical complexity of S. officinale, revealing not only numerous toxic PAs, but also a significant number of valuable phenolic compounds that could contribute to the bioactivities of comfreybased preparations

Lavandula stoechas subsp. luisieri is a Spanish subspecies from the Lamiaceae family. Its essential oil has been traditionally used for several medical applications though little is known about other extracts. Similar to many other studies aiming to obtain traditional plant extracts to be used in di_erent applications, this work evaluated the antioxidant and antimicrobial activities of Lavandula luisieri extracts and the correlation with their composition. Traditional hydrodistillation and ethanolic maceration were used to obtain the essential oil and the maceration extract, respectively. A green and sustainable methodology was applied to the maceration extract that was under a Supercritical Antisolvent Fractionation process to obtain a fine solid enriched in rosmarinic acid and the terpenes oleanolic and ursolic acids. Antimicrobial activities of all extracts and pure identified compounds (rosmarinic and ursolic acids) were evaluated against five bacterial strains; Listeria monocytogenes, Enterococcus faecium, Staphylococcus aureus, Salmonella Typhimurium and Escherichia coli and were compared with the pure compounds identified, rosmarinic and ursolic acids. All strains were sensitive against L. luisieri essential oil. The solid product obtained from the supercritical process was concentrated in the identified actives compared to the maceration extract, which resulted in higher antimicrobial and DPPH scavenging activities. The supercritical sustainable process provided L. luisieri compounds, with retention of their antimicrobial and antioxidant activities, in a poder exemptof organic solvents with potential application in the clinical, food or cosmetic fields., Fondo Europeo de Desarrollo Regional, Ministerio de Economía, Comercio y Empresa (España)/Fondo Europeo de Desarrollo Regional, Gobierno de Aragón/Fondo Europeo de Desarrollo Regional, Depto. de Química en Ciencias Farmacéuticas, Fac. de Farmacia, TRUE, pub

Excessive exposure to ultraviolet (UV) radiation is the main risk factor to develop skin pathologies or cancer because it encourages oxidative condition and skin inflammation. In this sense, strategies for its prevention are currently being evaluated. Natural products such as carotenoids or polyphenols, which are abundant in the marine environment, have been used in the prevention of oxidative stress due to their demonstrated antioxidant activities. Nevertheless, the anti-inflammatory activity and its implication in photo-prevention have not been extensively studied. Thus, we aimed to evaluate the combination of fucoxanthin (FX) and rosmarinic acid (RA) on cell viability, apoptosis induction, inflammasome regulation, and anti-oxidative response activation in UVB-irradiated HaCaT keratinocytes. We demonstrated for the first time that the combination of FX and RA (5 µM RA plus 5 µM FX, designated as M2) improved antioxidant and anti-inflammatory profiles in comparison to compounds assayed individually, by reducing UVB-induced apoptosis and the consequent ROS production. Furthermore, the M2 combination modulated the inflammatory response through down-regulation of inflammasome components such as NLRP3, ASC, and Caspase-1, and the interleukin (IL)-1β production. In addition, Nrf2 and HO-1 antioxidant genes expression increased in UVB-exposed HaCaT cells pre-treated with M2. These results suggest that this combination of natural products exerts photo-protective effects by down-regulating NRLP3-inflammasome and increasing Nrf2 signalling pathway.

In the present research, the chemical composition of aerial parts of cultivated citral chemotype of Thymus pannonicus All. (Lamiaceae), growing at different soil conditions was studied. Two plant lineages were unfertilized or treated using either nitrogen or phosphorus fertilizers at different concentration levels. Essential oil (EO) content, EO composition, and composition of hydromethanol extracts (MEs) were compared to the results obtained for wild growing plants in order to evaluate the potential of T. pannonicus for its industrial production. EO content (%, V/m) in cultivated samples varied between 0.32-0.75% and 0.62-1.05% within two examined lineages, whereas the herbs from the natural habitat contained 0.49-1.29% of EO. GC-FID/MS analysis showed that citral, i.e. mixture of isomers geranial and neral, was the most abundant constituent in all EOs (51.90-81.96%). HPLC analysis revealed that total caffeic acid derivatives, with rosmarinic acid (RA) as the most abundant compound in all extracts, as well as total apigenin glycosides contents were significantly higher in the extracts originating from the cultivated herbs (112.15-184.94 mg/g and 5.08-15.29 mg/g, respectively), in comparison to the extracts of the herbs from the natural habitats (67.61-98.75 mg/g and 1.17-1.32 mg/g). In general, the composition of cultivated herbs varied less than that of samples originating from natural habitats, indicating that controlled cultivation of citral chemotype of T. pannonicus can provide herbal drug with favourable characteristics. Only minor differences were observed between cultivated samples treated with different fertilizers.

Lead (Pb) is one of the toxic heavy metals that have several toxicological implications including cytotoxicities and oxidative stress. The release of reactive oxygen species (ROS) usually initiates lipid peroxidation and resulting in inflammation and tissue injury. However, the detailed identification of the Pb-produced lipid hydroperoxides has received little attention. Furthermore, the mechanisms behind such effects are less informed. Therefore, this study firstly investigated Pb-produced lipid hydroperoxides in human HepG2 cells using LC/MS. The effects of Pb on the antioxidant enzymes were additionally examined using qPCR and their dependent activities. As a protection trial, the ameliorative effects of rosmarinic (RMA) and ascorbic (ASA) acids on Pb-induced cytotoxicity and oxidative stress and their regulatory effects on Nrf2/Keap1 pathway were investigated. The achieved results confirmed cytotoxicity and oxidative damage of Pb on HepG2 cells. In addition, 20 lipid hydroperoxides (LOOH) were identified including 11 phosphatidylcholine hydroperoxides (PCOOH), 5 triacylglycerol hydroperoxides (TGOOH) and 4 cholesteryl ester hydroperoxides (CEOOH). The most dominant LOOH species were PCOOH 34:2, PCOOH 34:3, PCOOH 38:7, TGOOH 60:14, TGOOH 60:15, CEOOH 18:3 and CEOOH 20:4. Pb significantly downregulated Nrf2-regulated antioxidant enzymes at both the pretranscriptional and functional levels. Co-exposure of HepG2 cells to RMA and ASA significantly reduced Pb-produced adverse outcomes. This protection occurred via activation Nrf2-Keap1 antioxidant pathway.

This study was undertaken to estimate the concentrations of the formed polycyclic aromatic hydrocarbons (PAHs) in heat-treated (boiled, pan-fried and grilled) meats collected from Egypt. Dietary intakes and cancer risks of PAHs among Egyptian adults were calculated. Benzo[a]pyrene (B[a]P)-induced mutagenicity and oxidative stress in human colon (CaCo-2) cell line and mechanisms behind such effects were also investigated. Finally, protection trials using rosmarinic (RMA) and ascorbic acids (ASA) were carried out. The results indicated formation of PAHs at high levels in the heat-treated meats. Calculated incremental life time cancer risk among Egyptian adults were 7.05179E - 07, 7.00604 E - 06 and 1.86069 E - 05 due to ingestion of boiled, panfried and grilled meats, respectively. B[a]P-exposed CaCo-2 cells had high abilities for mutagenicity (490.05 +/- 21.37 His + revertants) and production of reactive oxygen species. RMA and ASA protected CaCo-2 cells via reduction of B[a]P-induced mutagenicity and oxidative stress and upregulation of phase II detoxification enzymes and xenobiotic transporters.

This study was undertaken to estimate the concentrations of the formed polycyclic aromatic hydrocarbons (PAHs) in heat-treated (boiled, pan-fried and grilled) meats collected from Egypt. Dietary intakes and cancer risks of PAHs among Egyptian adults were calculated. Benzo[a]pyrene (B[a]P)-induced mutagenicity and oxidative stress in human colon (CaCo-2) cell line and mechanisms behind such effects were also investigated. Finally, protection trials using rosmarinic (RMA) and ascorbic acids (ASA) were carried out. The results indicated formation of PAHs at high levels in the heat-treated meats. Calculated incremental life time cancer risk among Egyptian adults were 7.05179E - 07, 7.00604 E - 06 and 1.86069 E - 05 due to ingestion of boiled, panfried and grilled meats, respectively. B[a]P-exposed CaCo-2 cells had high abilities for mutagenicity (490.05 +/- 21.37 His + revertants) and production of reactive oxygen species. RMA and ASA protected CaCo-2 cells via reduction of B[a]P-induced mutagenicity and oxidative stress and upregulation of phase II detoxification enzymes and xenobiotic transporters.

Lead (Pb) is one of the toxic heavy metals that have several toxicological implications including cytotoxicities and oxidative stress. The release of reactive oxygen species (ROS) usually initiates lipid peroxidation and resulting in inflammation and tissue injury. However, the detailed identification of the Pb-produced lipid hydroperoxides has received little attention. Furthermore, the mechanisms behind such effects are less informed. Therefore, this study firstly investigated Pb-produced lipid hydroperoxides in human HepG2 cells using LC/MS. The effects of Pb on the antioxidant enzymes were additionally examined using qPCR and their dependent activities. As a protection trial, the ameliorative effects of rosmarinic (RMA) and ascorbic (ASA) acids on Pb-induced cytotoxicity and oxidative stress and their regulatory effects on Nrf2/Keap1 pathway were investigated. The achieved results confirmed cytotoxicity and oxidative damage of Pb on HepG2 cells. In addition, 20 lipid hydroperoxides (LOOH) were identified including 11 phosphatidylcholine hydroperoxides (PCOOH), 5 triacylglycerol hydroperoxides (TGOOH) and 4 cholesteryl ester hydroperoxides (CEOOH). The most dominant LOOH species were PCOOH 34:2, PCOOH 34:3, PCOOH 38:7, TGOOH 60:14, TGOOH 60:15, CEOOH 18:3 and CEOOH 20:4. Pb significantly downregulated Nrf2-regulated antioxidant enzymes at both the pretranscriptional and functional levels. Co-exposure of HepG2 cells to RMA and ASA significantly reduced Pb-produced adverse outcomes. This protection occurred via activation Nrf2-Keap1 antioxidant pathway.

In the present research, the chemical composition of aerial parts of cultivated citral chemotype of Thymus pannonicus All. (Lamiaceae), growing at different soil conditions was studied. Two plant lineages were unfertilized or treated using either nitrogen or phosphorus fertilizers at different concentration levels. Essential oil (EO) content, EO composition, and composition of hydromethanol extracts (MEs) were compared to the results obtained for wild growing plants in order to evaluate the potential of T. pannonicus for its industrial production. EO content (%, V/m) in cultivated samples varied between 0.32-0.75% and 0.62-1.05% within two examined lineages, whereas the herbs from the natural habitat contained 0.49-1.29% of EO. GC-FID/MS analysis showed that citral, i.e. mixture of isomers geranial and neral, was the most abundant constituent in all EOs (51.90-81.96%). HPLC analysis revealed that total caffeic acid derivatives, with rosmarinic acid (RA) as the most abundant compound in all extracts, as well as total apigenin glycosides contents were significantly higher in the extracts originating from the cultivated herbs (112.15-184.94 mg/g and 5.08-15.29 mg/g, respectively), in comparison to the extracts of the herbs from the natural habitats (67.61-98.75 mg/g and 1.17-1.32 mg/g). In general, the composition of cultivated herbs varied less than that of samples originating from natural habitats, indicating that controlled cultivation of citral chemotype of T. pannonicus can provide herbal drug with favourable characteristics. Only minor differences were observed between cultivated samples treated with different fertilizers.

In this dissertation, the influence of diamagnetic metal ions Mg (II) and Ca (II) on the autoxidation of polyphenol compounds, containing one or two ortho-dihydroxyl groups, in aqueous solutions at different pH values, was studied. The autoxidation of polyphenols distributed in nature, such as: catechol, caffeic acid, rosmarinic acid, chlorogenic acid, rutin and hematoxylin was investigated. For this purpose, the following instrumental methods were applied: UV/Vis spectrophotometry with subsequent hemometric treatment of UV/Vis spectra, ESR spectroscopy, high performance liquid chromatography (HPLC) and mass spectrometry (MS). It has been shown that the pH value of the solution and/or the presence and type of metal ions affect the speed and manner of the autoxidation processes of the investigated compounds. Applied techniques have enabled the determination of the number and structure of intermediate products as well as stable autoxidation products. The obtained results allow a more detailed insight into mechanisms of antioxidative activities of studied polyphenols and a more comprehensive view of their biochemical behavior with the implications for further human use.

During the essential oil steam distillation from aromatic herbs, huge amounts of distillation wastewaters (DWWs) are generated. These by-products represent an exceptionally rich source of phenolic compounds such as rosmarinic acid (RA) and caffeic acid (CA). Herein, the alternative use of dried basil DWWs (dDWWs) to perform a selective extraction of RA and CA by pressurized liquid extraction (PLE) employing bio-based solvent was studied. To select the most suitable solvent for PLE, the theoretical modelling of Hansen solubility parameters (HSP) was carried out. This approach allows reducing the list of candidate to two solvents: ethanol and ethyl lactate. Due to the composition of the sample, mixtures of water with those solvents were also tested. An enriched PLE extract in RA (23.90 ± 2.06 mg/g extract) with an extraction efficiency of 75.89 ± 16.03% employing a water-ethanol mixture 25:75 (% v/v) at 50°C was obtained. In the case of CA, a PLE extract with 2.42 ± 0.04 mg/g extract, having an extraction efficiency of 13.86 ± 4.96% using ethanol absolute at 50°C was achieved. DWWs are proposed as new promising sources of natural additives and/or functional ingredients for cosmetic, nutraceutical, and food applications.

During the essential oil steam distillation from aromatic herbs, huge amounts of distillation wastewaters (DWWs) are generated. These by‐products represent an exceptionally rich source of phenolic compounds such as rosmarinic acid (RA) and caffeic acid (CA). Herein, the alternative use of dried basil DWWs (dDWWs) to perform a selective extraction of RA and CA by pressurized liquid extraction (PLE) employing bio‐based solvent was studied. To select the most suitable solvent for PLE, the theoretical modelling of Hansen solubility parameters (HSP) was carried out. This approach allows reducing the list of candidate to two solvents: ethanol and ethyl lactate. Due to the composition of the sample, mixtures of water with those solvents were also tested. An enriched PLE extract in RA (23.90 ± 2.06 mg/g extract) with an extraction efficiency of 75.89 ± 16.03% employing a water‐ethanol mixture 25:75 (% v/v) at 50°C was obtained. In the case of CA, a PLE extract with 2.42 ± 0.04 mg/g extract, having an extraction efficiency of 13.86 ± 4.96% using ethanol absolute at 50°C was achieved. DWWs are proposed as new promising sources of natural additives and/or functional ingredients for cosmetic, nutraceutical, and food applications.

[Background]: Rosmarinus officinalis is an aromatic plant used in folk medicine as a result of the therapeutic properties associated with its phenolic composition, being rich in rosmarinic acid (RA) and caffeic acid (CA). To better understand the bioactivity of these compounds, their absorption and metabolism were assessed in human Caco-2 and HepG2 cells, as small intestine and liver models, respectively, using RA and CA standards, as well as a rosemary infusion and ferulic acid (FA). [Results]: Test compounds were partially up-taken and metabolized by Caco-2 and HepG2 cells, although a higher metabolization rate was observed after hepatic incubation compared to intestinal incubation. CA was the compound best absorbed followed by RA and FA, showing metabolites percentages of 30.4%, 11.8% and 4.4% in Caco-2 and 34.3%, 10.3% and 3.2% in HepG2 cells, respectively. RA in the rosemary infusion showed improved bioavailability compared to pure RA. Methyl derivatives were the main metabolites detected for CA and RA after intestinal and hepatic metabolism, followed by methyl-glucuronidates and glucuronidates. RA was also minimally hydrolyzed into CA, whereas FA only was glucuronidated. Rosemary polyphenols followed the same biotransformation pathways as the standards. In addition, phase II derivatives of luteolin were observed. [Conclusion]: Rosemary polyphenols are partially metabolized in both the intestine and liver.

Amyloid-β aggregation inhibitors are expected to be therapeutic or prophylactic agents for Alzheimer's disease. Rosmarinic acid, which is one of the main aggregation inhibitors derived from Lamiaceae, was employed as a lead compound and its 25 derivatives were synthesized. In this study,the structure-activity relations of rosmarinic acid derivatives for the amyloid-β aggregation inhibitory effect (MSHTS assay), antioxidant properties, and xanthine oxidase inhibition were evaluated. Among the tested compounds, compounds 16d and 19 were found to the most potent amyloid aggregation inhibitors. The SAR revealed that the necessity of the presence of the phenolic hydroxyl on one side of the molecule as well as the lipophilicity of the entire molecule. The importance of these structural properties was also supported by docking simulations.

The presented study aimed to investigate the elicitation possibility for the production of main secondary metabolites in Nepeta cataria L. and N. pannonica L. plants, by exposing them to synthetic compounds belonging to tetraoxanes and thiophenes group. The effect of DO63 (1,2,4,5-tetraoxane) and DOVF15 (2,5-diphenylthiophene) on the production of cis-trans-nepetalactone (NL) and rosmarinic acid (RA) in two Nepeta species, was investigated in shoots grown on the culture medium with the addition of synthetic compounds in the concentrations ranging from 0.1 to 2 mg L-1. The content of targeted metabolites in tested in vitro shoots depended on the type and concentration of applied synthetic compounds. Application of DO63 in the concentration range 0.1-1 mg L-1 affected only NL production in both Nepeta species resulting in its increased content, while production of RA was not influenced in the treated shoots. Addition of DOVF15 caused decreased RA content in N. pannonica shoots and an increase in N. cataria shoots, whereas NL production was not affected. The presented results reveal the possibility of DO63 and DOVF15 application for the elicitation of the main secondary metabolites production in species from the genus Nepeta.

The presented study aimed to investigate the elicitation possibility for the production of main secondary metabolites in Nepeta cataria L. and N. pannonica L. plants, by exposing them to synthetic compounds belonging to tetraoxanes and thiophenes group. The effect of DO63 (1,2,4,5-tetraoxane) and DOVF15 (2,5-diphenylthiophene) on the production of cis-trans-nepetalactone (NL) and rosmarinic acid (RA) in two Nepeta species, was investigated in shoots grown on the culture medium with the addition of synthetic compounds in the concentrations ranging from 0.1 to 2 mg L-1. The content of targeted metabolites in tested in vitro shoots depended on the type and concentration of applied synthetic compounds. Application of DO63 in the concentration range 0.1-1 mg L-1 affected only NL production in both Nepeta species resulting in its increased content, while production of RA was not influenced in the treated shoots. Addition of DOVF15 caused decreased RA content in N. pannonica shoots and an increase in N. cataria shoots, whereas NL production was not affected. The presented results reveal the possibility of DO63 and DOVF15 application for the elicitation of the main secondary metabolites production in species from the genus Nepeta.

3P-259

An aqueous extract of the root of comfrey (Symphytum officinale L.) was obtained by the Soxlet method using water as an extractant. Extraction with water was carried out at predetermined optimal conditions of extraction (extraction time 240min and hydromodule 1:10w/v). For modeling the kinetics of extraction of total extractable compounds from comfrey root with water, Ponomarev’s kinetic model was applied wherein extraction degree (SE) in the period of rapid extraction and coefficients b and k in the equations of extraction kinetics were determined. Chemical composition of an aqueous extract of comfrey root obtained under optimal conditions was examined. The content of total phenols (473.16mg GKE/g d.e.) and flavonoids (18.90 mg RE/g d.e.) was determined. HPLC, UHPLC-DAD-HESI-MS and ICP-OES methods for identification and quantification of bioactive compounds and micro and macro elements present in the aqueous extract of comfrey root were validated. The most abundant bioactive compounds in the examined aqueous extract of comfrey root were rosmarinic acid, allantoin and ellagic acid (7.4g/100g d.e, 8.91g/100g d.e and 12.891g/100 g s.e), respectively. The mineral composition of an aqueous extract of comfrey root showed that there was a relatively high content of K (560,4mg/dm3) and Ca (129,3mg/dm3). Other trace elements contained in an aqueous extract of comfrey root in descending values were Na, Pb, Mg, Fe, Al, Zn, B, Ba, Cu. Examination of the effects of aqueous comfrey root extract included the examination of antioxidant activity by monitoring the ability of neutralization of free radicals and antimicrobial activity in vitro in parallel with the standards of bioactive compounds that are the most present in the extract. Obtained EC50 values for tested samples showed that the highest antioxidant activity have ellagic, then rosmarinic acid, and finally aqueous extract of comfrey root, while allantoin did not show antioxidant activity. The aqueous extract of comfrey roo

We investigated the effect of the natural polyphenols rosmarinic acid and arbutin on osteoclast differentiation in RAW 264.7 cells. Rosmarinic acid and arbutin suppressed osteoclast differentiation and had no cytotoxic effect on osteoclast precursor cells. Rosmarinic acid and arbutin inhibited superoxide production and intracellular superoxide in a dose dependent manner. mRNA expression of the master regulator of osteoclastogenesis, nuclear factor of activated T cells cytoplasmic1(NFATc1), and the osteoclast marker genes, matrix metalloproteinase -9, tartrate-resistant acid phosphatase and cathepsin-K, decreased after treatments with rosmarinic acid and arbutin. Furthermore, resorption activity decreased with the number of osteoclasts. These results suggest that rosmarinic acid and arbutin may be useful for the prevention and treatment of bone diseases such as osteoporosis through mechanisms involving inhibition of superoxide and down-regulation of NFATc1.

Hydroxycinnamic acids undoubtedly contribute to the wide range of pharmacological effects of the species of Thyme genus. Rosmarinic acid has antioxidant, hepatoprotective, cardioprotective, nephro-protective, anti-inflammatory, immunomodulatory, anti-allergic and anti-tumor properties. Caffeic acid has the antimicrobial, fungistatic activity, produces a choleretic effect. Accumulation of these acids is specific for Lamiaceae family. The aim of our research was to determine the type of hydroxycinnamic acids, the concentration of rosemarinic and caffeic acids and the total content of hydroxycinnamic acids, also to determine parameters of perfect extraction of this type of biological active compounds in the herb of four species of Thyme genus of the European flora. Identification of hydroxycinnamic acids has been carried out using paper and thin layer chromatography in different systems of solvents in comparison with standard compounds. Using TLC and HPLC caffeic and rosmarinic acids have been identified in the herb samples of Thymus Marshallianus, Thymus pulegioides, Thymus crenulatus, Thymus dimorphus. The results of HPLC analysis of caffeic and rosmarinic acids in the herb of Thymus species show that the concentration of rosmarinic acid ranges from 2343.40 mg/kg to 14351.74 mg/kg; the content of caffeic acid ranges from 74.41 mg/kg to 93.86 mg/kg. The highest concentration of rosmarinic acid, as well as caffeic acid was in Thymus pulegioides. The total content of hydroxycinnamic acids has been determined by spectrophotometry, and it ranges from 3.27 to 19.28%. Thymus crenulatus is the richest species by accumulation of the total content hydroxycinnamic acids. The optimal parameters for herb extraction such as the size of the crushed herb particles, extragent and extraction time have been found., Гидроксикоричные кислоты несомненно обусловливают широкий спектр фармакологической активности растений рода тимьян. Розмариновая кислота проявляет антиоксидантные, гепатопротекторные, кардиопротекторные, нефропротекторные, противовоспалительные, иммуномодулирующие, противоаллергические и противоопухолевые свойства. Кофейная кислота обладает противомикробным, фунгистатическим и желчегонным действием. Накопление этих кислот специфично для растений семейства Яснотковые. Целью исследований было определение производных гидроксикоричной кислоты, содержания розмариновой и кофейной кислот, а также суммы гидроксикоричных кислот, анализ параметров оптимальной экстракции этого класса биологически активных соединений травы четырех видов рода тимьян европейской флоры. Идентификацию производных гидроксикоричной кислоты проводили с помощью бумажной и тонкослойной хроматографии в ряде систем растворителей в сравнении с достоверными веществами. С помощью ТСХ и ВЭЖХ анализа идентифицировали кофейную и розмариновую кислоты в траве видов тимьяна Thymus, Thymus pulegioides, Thymus crenulatus, Thymus dimorphus. Результаты ВЭЖХ анализа показывают, что содержание розмариновой кислоты варьирует от 2343,40 мг/кг до 14351,74 мг/кг, а кофейной – от 74,41 мг/кг до 93,86 мг/кг. Самое высокое содержание розмариновой кислоты, так же как и кофейной было характерно для травы Thymus pulegioides. Содержание суммы гидроксикоричных кислот устанавливали спектрофотометрически и оно составляло 3,27-19,28%. Наивысший уровень суммы гидроксикоричных кислот наблюдался в траве Thymus crenulatus. Определены оптимальные параметры экстракции травы тимьяна: размер частиц, тип экстрагента и время экстракции., Гідроксикоричні кислоти обумовлюють широкий спектр фармакологічної активності рослин роду чебрець. Розмаринова кислота продукує антиоксидантну, гепатопротекторну, кардіопротекторну, нефропротекторну, протизапальну, імуномодулюючу, протиалергічну та протипухлинні властивості. Кофейна кислота чинить антимікробну, фунгістатичну, жовчогінну дію. Накопичення цих кислот специфічне для рослин родини Ясноткові. Метою дослідження було визначення похідних гідроксикоричної кислоти, концентрації розмаринової та кофейної кислот, а також суми гідроксикоричних кислот, визначення параметрів досконалої екстракції цього класу біологічно активних сполук у траві чотирьох видів роду чебрець європейської флори. Ідентифікацію похідних гідроксикоричної кислоти визначали за допомогою паперової та тонкошарової хроматографії у ряду систем розчинників у порівнянні з вірогідними речовинами. За допомогою ТШХ і ВЕРХ аналізу ідентифікували кофейну і розмаринову кислоти у траві видів чебрецю Thymus, Thymus pulegioides, Thymus crenulatus, Thymus dimorphus. Результати ВЕРХ аналізу показують, що вміст розмаринової кислоти варіює від 2343,40 мг/кг до 14351,74 мг/кг, а кофейної – від 74,41 мг/кг до 93,86 мг/кг. Найвища концентрація розмаринової кислоти, також як і кофейної була відзначена у траві Thymus pulegioides. Вміст суми гідроксикоричних кислот встановлювали спектрофотометричним методом і він складав 3,27-19,28%. Найвищий рівень суми гідроксикоричних кислот спостерегається у траві Thymus crenulatus. Визначені оптимальні параметри екстракції трави чебрецю: розмір частинок, тип екстрагенту і час екстракції.

Plants are an important source for medicinal compounds. Chemical screening and selection is critical for identification of compounds of interest. Ocimum basilicum (Basil) is a rich source of polyphenolics and exhibits high diversity, therefore bioprospecting of a suitable cultivar is a necessity. This study reports on the development of a true to type novel "in vitro system" and its comparison with a conventional system for screening and selection of cultivars for high total phenolics, individual polyphenolics, and antioxidant content. We have shown for the first time using online acidic potassium permanganate chemiluminescence that extracts from Ocimum basilicum showed antioxidant potential. The current study identified the cultivar specific composition of polyphenolics and their antioxidant properties. Further, a distinct relationship between plant morphotype and polyphenolic content was also found. Of the 15 cultivars examined, "Holy Green", "Red Rubin", and "Basil Genovese" were identified as high polyphenolic producing cultivars while "Subja" was determined to be a low producer. The "in vitro system" enabled differentiation of the cultivars in their morphology, polyphenolic content, and antioxidant activity and is a cheap and efficient method for bioprospecting studies.

Borage (Borago officinalis L.) is a typical Spanish plant. During processing, 60% are leaves. The aim of this work is to model and optimize the extraction of polyphenol from borage leaves using the response surface method (RSM) and to use this extract for application in emulsions. The responses were: total polyphenol content (TPC), antioxidant capacity by ORAC, and rosmarinic acid by HPLC. The ranges of the variables temperature, ethanol content and time were 50–90 °C, 0%–30%–60% ethanol (v/v), and 10–15 min. For ethanolic extraction, optimal conditions were at 75.9 °C, 52% ethanol and 14.8 min, yielding activity of 27.05 mg GAE/g DW TPC; 115.96 mg TE/g DW in ORAC and 11.02 mg/L rosmarinic acid. For water extraction, optimal activity was achieved with extraction at 98.3 °C and 22 min, with responses of 22.3 mg GAE/g DW TPC; 81.6 mg TE/g DW in ORAC and 3.9 mg/L rosmarinic acid. The significant variables were ethanol concentration and temperature. For emulsions, the peroxide value was inhibited by 60% for 3% extract concentration; and 80% with 3% extract concentration and 0.2% of BSA. The p-anisidine value between the control and the emulsion with 3% extract was reduced to 73.6% and with BSA 86.3%, and others concentrations had similar behavior., Postprint (published version)

目的:研究腫節風藥材的高效液相色譜(HPLC)指紋圖譜,并測定藥材中異嗪皮啶與迷迭香酸的含量,以建立藥材有效的質量評價方法。方法:色譜柱為Inertsil ODS-4(250 mm×4.6 mm,5μm),流動相為0.1%磷酸溶液-乙腈(梯度洗脫),流速為1.0 ml/min,檢測波長為342 nm。結果:所得腫節風藥材的HPLC指紋圖譜分離度、精密度、重復性和穩定性均良好;10批腫節風藥材指紋圖譜的平均相似度為0.853,不同產地樣品的指紋圖譜差異不大,但4個代表性色譜峰的峰面積有所區別。異嗪皮啶與迷迭香酸的質量濃度均在5~500 mg/ml范圍內與各自峰面積積分值呈良好的線性關系(r分別為0.999 5、0.998 9);平均加樣回收率分別為105.20%、99.52%,RSD分別為1.39%、1.56%(n均為6);10批腫節風藥材中異嗪皮啶與迷迭香酸的含量存在明顯差異,尤以迷迭香酸含量差異較大。結論:該方法簡便、準確、重復性好,可以作為腫節風藥材的質量控制方法。 OBJECTIVE：To study HPLC fingerprints of Sarcandra glabra，and to determine the contents of isofraxidin and rosmarinic acid and establish the method for quality evaluation. METHODS：The samples were separated by Inertsil ODS-4（250 mm× 4.6 mm，5 μm）column with mobile phase consisted of 0.1％ phosphoric acid-acetonitrile（gradient elution）at the flow rate of 1.0 ml/ min. The detection wavelength was set at 342 nm. RESULTS：HPLC fingerprint of S. glabra was established with good separation， precision，repeatability and stability；average similarity of 10 batches of S. glabra was 0.853；there was no significant difference in fingerprint of samples from different producing areas，but the contents of 4 representative peaks were different from one another. The linear ranges of isofraxidin were 5-500 mg/ml（r＝0.999 5）and rosmarinic acid were 5-500 mg/ml（r＝0.998 9）. Average recoveries were 105.20％（RSD＝1.39％ ，n＝6）and 99.52％（RSD＝1.56％ ，n＝6）. The contents of isofraxidin and rosmarinic acid from 10 batches were obviously different from one another，especially rosmarinic acid. CONCLUSIONS：The method is simple，accurate and repeatable，which can be used for the quality control of S. glabra.

Rindera umbellata је ретка и крајње угрожена биљна врста у Србији. Да би се успоставили протоколи за ex situ заштиту и обезбедио материјал за молекуларне и фитохемијске анализе успостављена је in vitro култура ове слабо испитане врсте. Оптимизација услова гајења заснивала се на варирању врсте и концентрације угљених хидрата (сахароза, глукоза и фруктоза) и фитохормона (гиберелна и јасмонска киселина) у подлози за гајење. Такође је испитиван утицај наведених регулатора растења на продукцију фармацеутски важних секундарних метаболита. In vitro растење и развиће биљака врсте R. umbellata је под утицајем како врсте, тако и концентрације примењеног шећера. Утврђено је да сахароза представља најбољи извор угљеника за in vitro раст, ожиљавање и аклиматизацију биљака, а да су концентрације од 0,3 и 1 М свих примењених шећера инхибиторне. Гиберелна киселина стимулише раст и издуживање стабла ових розетастих биљака, али не утиче значајно на процес ожиљавања. Јасмонска киселина је стимулисала in vitro раст и ожиљавање R. umbellatа, нарочито при нижим концентрацијама и на краткотрајним третманима. Највиша примењена концентрација овог фитохормона (200 μМ) инхибирала је све посматране процесе. Уочена су значајна варирања у квантитативном и квалитативном садржају секундарних метаболита зависно од начина гајења, фенофазе и тестираног биљног органа. Биљке врсте R. umbellata у природним условима синтетишу незасићене и засићене пиролизидинске алкалоиде. Биљке гајене in vitro синтетишу веома ниске концентрације искључиво засићених пиролизидинских алкалоида. Фенолна једињења детектована су у листовима и корену ових биљака. Од фенолних киселина биљке из природе садрже више рузмаринске киселине у надземним деловима, док је садржај литоспермичне киселине Б у кореновима вишеструко већи од количине рузмаринске киселине. Сличан однос садржаја ових једињења запажен је и у биљкама које су гајене у in vitro култури. Иако су највише примењене концентрације сахарозе биле инхибиторне за растење, би, Rindera umbellata is a rare and critically endangered plant species in Serbia. In order to optimize protocols for ex situ conservation and provide the materials for molecular and phytochemical analysis, a suitable method for in vitro propagation of this plant species was established. Optimization of the growing conditions was based on varying the type and concentration of carbohydrates (sucrose, glucose and fructose) and phytohormones (gibberellic acid and jasmonic acid) in the substrate. Furthermore, the effect of selected growth regulators on the production of pharmaceutically important secondary metabolites was investigated. In vitro growth and development of R. umbellata plants were significantly affected by both the type and the concentration of applied sugars. It was found that sucrose is the best carbon source for in vitro growth, rooting and acclimatization of the plants, and that the concentrations of 0.3 and 1 M of all applied sugars had an inhibitory effect. Gibberellic acid stimulated growth and stem elongation, but did not significantly affect the process of rooting. Jasmonic acid stimulated in vitro growth and rooting of R. umbellata, especially at lower concentrations, and after short-term treatments. Significant variations in quantitative and qualitative contents of secondary metabolites are noticed, which depended on the method of cultivation, growth stages and plant organs tested. Plants of R. umbellata grown under natural conditions synthesized both unsaturated and saturated pyrrolizidine alkaloids. In vitro grown plants synthesized a very low concentration of saturated alkaloids. Phenol compounds were detected in the leaves and roots of the plants. The content of lithospermic acid B in the roots was several times greater than the amount of rosmarinic acid. A similar relation between the contents of rosmarinic acid and lithospermic acid B was observed in plants that were grown in vitro. Gibberellic acid did not affect the production of phenolic co

The low incidence of cardiovascular disease in Mediterranean countries leads to an increased interest of the scientific community for the Mediterranean diet. Our aim was to evaluate total phenol and flavonoid contents, antioxidant capacity, free radical scavenging activity and potential antihypertensive effect of aqueous extract obtained from Thymus serpyllum L. (wild thyme, TE), an aromatic herb from the Lamiaceae family (highly present in Mediterranean diet), in spontaneously hypertensive rats (SHR) and in normotensive Wistar rats. Total phenol content of TE was 2008.33 +/- 10.6 mg/L GAE, and rosmarinic and caffeic acids were predominant phenolic compounds. The ferric reducing/antioxidant power and antioxidant capacity analysis revealed strong antioxidative properties of TE. In vitro nitric oxide-scavenging activity of 1 mg/l TE was 63.43 % with the IC50 value of 122.36 mu g/ml. Bolus injection of TE (100 mg/kg body weight i.v.) induced significant decrease of systolic and diastolic blood pressure and total peripheral resistance in SHR, without effects on these parameters in normotensive Wistar rats. Cardiac index remained unchanged after TE treatment in all experimental rats. Given dose of TE did not show significant nitric oxide-scavenging activity in vivo. Our results indicate that TE may protect against hypertension in experimental model of essential hypertension.

Methanolic extract of Satureja kitaibelii Wierzb. ex Heuff. (Lamiaceae) and its major compond rosmarinic acid possessed strong antimicrobial activity against tested bacteria and fungi, and antitumour activity against malignant cells, but without antiproliferative effect on healthy cell line MRC5. Minimal inhibitory concentration (MIC) of methanolic extract was ranged between 0.0125-1.25 mg/ml, while minimal bactericidal concentration (MBC) was of 0.2-5.0 mg/ml and fungicidal (MFC) was 0.15-2.5 mg/ml. The rosmarinic acid exhibited better antimicrobial activity then methanol extract. The most sensitive species to rosmarinic acid were Bacillus cereus and Candida kruzei, while Micrococcus flavus, Trichoderma viride and Aspergillus niger were the most resistant. The methanol extract exhibited strong activity against Fem-x human malignant melanoma cells with an IC50 39.66 +/- 2.71 mu g/ml, and moderate activity against other cancer cell lines (IC50 from 138.06 +/- 0.16 mu g/ml against estrogen-dependant breast cancer cell lines MDA-MB-361 to 173.15 +/- 0.02 against a human epithelial cervical cancer cells HeLa). Rosmarinic acid arrested G2/M phase cell cycle in Fem-x cells, against which both methanol extract and rosmarinic acid possessed the best cytotoxic activity.

The low incidence of cardiovascular disease in Mediterranean countries leads to an increased interest of the scientific community for the Mediterranean diet. Our aim was to evaluate total phenol and flavonoid contents, antioxidant capacity, free radical scavenging activity and potential antihypertensive effect of aqueous extract obtained from Thymus serpyllum L. (wild thyme, TE), an aromatic herb from the Lamiaceae family (highly present in Mediterranean diet), in spontaneously hypertensive rats (SHR) and in normotensive Wistar rats. Total phenol content of TE was 2008.33 +/- 10.6 mg/L GAE, and rosmarinic and caffeic acids were predominant phenolic compounds. The ferric reducing/antioxidant power and antioxidant capacity analysis revealed strong antioxidative properties of TE. In vitro nitric oxide-scavenging activity of 1 mg/l TE was 63.43 % with the IC50 value of 122.36 mu g/ml. Bolus injection of TE (100 mg/kg body weight i.v.) induced significant decrease of systolic and diastolic blood pressure and total peripheral resistance in SHR, without effects on these parameters in normotensive Wistar rats. Cardiac index remained unchanged after TE treatment in all experimental rats. Given dose of TE did not show significant nitric oxide-scavenging activity in vivo. Our results indicate that TE may protect against hypertension in experimental model of essential hypertension.

The search of new natural sources of bioactive compounds to be employed in the food industry is a hot topic in the field of functional foods. Besides, the use of environmentally respectful technologies focused on the extraction of bioactives from natural matrices is of great importance nowadays. Pressurized liquid extraction (PLE) is a novel extraction method that allows obtaining higher extraction efficiencies while minimizing both, the amount of solvents and the time. On the other hand, enzyme assisted extraction (EAE) can be seen as a way to further improve the extraction mass transfer kinetics when dealing with plant-based materials. In fact, the use of enzymes to break down cells walls might offer new possibilities because the potential bioactive compounds found in cells will be more available to the extraction solvent. In these work, the two extraction mechanisms are studied and tested for the extraction of natural antioxidant compounds from lemon balm (Melissa officinalis). Different PLE extraction conditions are tested using ethanol or water as extracting solvents. Besides, the use of different enzymes, namely cellulose, xylanase and pectinase, alone or in combination, is also studied. Best results in terms of total antioxidant activity and extraction yield were obtained when using PLE using water as extracting solvent at 150 °C. Besides, the obtained extracts were chemically characterized using an HPLC-DAD-MS method. Thanks to the combination of these techniques, several interesting phenolic compounds could be identified in the obtained extracts, among which rosmarinic acid is highlighted.

In an attempt to understand the beneficial health effects of Artemisia annua other than its anti-malaria properties, extracts from different cultivars prepared as tea infusions were investigated using Caco-2 cells on the intestinal inflammation and cytochrome P450 (CYP) activities. The characterisation of their phenolic compound (PC) profile revealed rosmarinic and chlorogenic acids as the main PCs. The extracts, assayed on Caco-2 cells at a plausible intestinal concentration, significantly decreased the secretion of pro-inflammatory cytokines, IL-8 and IL-6. This effect could be attributable at least to their content in rosmarinic acid, detected as a potent anti-inflammatory compound. The extracts also inhibited the activity of CYP3A4, whose expression was induced by 1,25-dihydroxyvitamin D3, and of CYP1A1, induced by benzo(a)pyrene. Our results highlight the advantage of drinking A. annua infusions for their potent anti-inflammatory effect, linked to PC content, which could synergise their antimalarial activity.