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With the proposal of the 2030 Agenda for Sustainable Development, the Chinese medicine extraction technology has been innovatively improved to prioritize low energy consumption, sustainability, and minimized organic solvent utilization. Forsythia suspensa (FS) possesses favorable pharmacological properties and is extensively utilized in traditional Chinese medicine. However, due to the limitations of the composition and extraction methods, its potential has not been fully developed. Thus, a combination of ultrasound-assisted extraction (UAE), enzyme-assisted extraction (EAE), and β-cyclodextrin extraction (β-CDE) was employed to isolate and purify rutin, phillyrin, and forsythoside A from FS. The results demonstrated that the efficiency of extracting enzymatic and ultrasound assisted β-cyclodextrin extraction (EUA-β-CDE) was highly influenced by the temperature and duration of hydrolysis, as well as the duration of the extraction process. According to the results of the single-factor experiment, Box-Behnken design (BBD) in Response surface method (RSM) was used to optimize the experimental parameters to achieve the maximum comprehensive evaluation value (CEV) value. The EUA-β-CDE compared with other extraction methods, has good extraction effect and low energy consumption by high performance liquid chromatography (HPLC), scanning electron microscopy (SEM), calculation of power consumption and CO2 emission The EUA-β-CDE compared with other extraction methods, has good extraction effect and low energy consumption by HPLC, SEM, calculation of power consumption and CO2 emission. Then, the structural characteristics of EUA-β-CDE of FS extract had significant interaction with β-CD by Fourier infrared spectroscopy (FT-IR) and differential scanning calorimetry (DSC). In addition, EUA-β-CDE extract has good antioxidant and anti-inflammatory activities. The establishment of EUA-β-CDE of FS provides a new idea for the development and application of other sustainable extraction methods of traditional Chinese medicine.

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Weeping forsythia (Forsythia suspensa, Oleaceae) is a deciduous broad-leaved tree species distributed in the warm temperate zone of China. However, the species still lacks a chromosome-level genome. In this study, the former draft genome (Accession No. WIPI00000000) of weeping forsythia was assembled into 14 chromosomes with a 712.9 Mb genome size. Weeping forsythia underwent α and β whole-genome duplication events. After the divergence between weeping forsythia and Olea europaea, 1 453 gene families had a significant expansion, and 1 146 gene families had a significant contraction. The enrichment pathways and ontologies of expanded genes suggested that the tillering, photosynthesis and growth capacity of weeping forsythia were enhanced after the divergence of weeping forsythia and O. europaea. The contracted genes suggested that the resistance of weeping forsythia to cold and drought was weakened. The last glacial period led to a significant decline in the effective population size of weeping forsythia. Forty-six candidate genes were identified for the synthesis of the forsythin and forsythoside A by genomic and transcriptomic data. In this study, we improved the previous draft genome of weeping forsythia. Our genome will provide genomic resources for the subsequent evolution and breeding research of weeping forsythia.

Forsythia suspensa (Thunb.) Vahl (FS) is an important plant with high edible and medicinal values. The edible fruit of FS is a common traditional medicine in China, Japan and Korea. Compared to the research on the phytochemistry and pharmacology of the fruits, study on the other parts of FS, such as leaves and flowers is still limited. In this study, an integrated strategy based on ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF-MS/MS), plant metabolomics and correlation analysis was established for comprehensively chemical characterization of fruits, leaves and flowers of FS, and discovery of α-glucosidase and pancreatic lipase inhibitory metabolites. The plant metabolic profiling of fruits, leaves and flowers of FS was performed by UPLC-QTOF-MS/MS, and a total of 74 secondary metabolites, including 15 phenylethanoid glycosides, 20 lignans, 10 cyclohexanol derivatives, 11 organic acids, 9 flavonoids, 3 triterpenes, and 10 other compounds were identified. Then, 29 differential secondary metabolites that responsible to distinguish the fruits, leaves and flowers of FS were further screened out by multivariate statistical analysis. Meanwhile, the α-glucosidase and pancreatic lipase inhibition of different parts of FS were evaluated and compared by in vitro experiments. The results demonstrated that the leaves of FS showed the highest inhibition on α-glucosidase and pancreatic lipase with IC50 of 0.17 ± 0.04 mg/mL and 0.56 ± 0.33 mg/mL, respectively. Then, the correlation between differential metabolites and enzyme inhibitory activities were investigated by Pearson correlation analysis, and 12 potential α-glucosidase and pancreatic lipase inhibitors were screen out. Additionally, the α-glucosidase and pancreatic lipase inhibitory activities of five potential enzyme inhibitory components, including quercitrin, rutin, kaempferol-3-O-rutinoside, pinoresinol-4-O-glucoside and phillyrin were further validated by in vitro assays and molecular docking analysis. The results showed that all the five potential inhibitors showed good inhibitory effects on α-glucosidase and pancreatic lipase, and the binding of the five inhibitors to enzymes mainly through hydrogen bonding, hydrophobic force, and ionic bonding. This study provided a feasible strategy for comparison and discrimination of different parts of medicinal plant and discovery of bioactive components, and also provided useful information for future utilization of different parts of FS.

Besides active substances, Forsythia suspensa is rich in dietary fiber (DF), but it is often wasted or discarded and not put to good use. In order to improve the function of Forsythia DF, it was modified using alkaline hydrogen peroxide (AHP) and cellulase (EM). Compared to the control DF (ODF), the DF modified using AHP (AHDF) and EM (EMDF) had a looser microstructure, lower crystallinity, and higher oil holding capacity (OHC) and cation exchange capacity (CEC). The AHP treatment significantly increased the water holding capacity (WHC) and water swelling ability (WSA) of the DF, while the EM treatment achieved just the opposite. Moreover, the functional properties of AHDF and EMDF, including their cholesterol adsorption capacity (CAC), nitrite ion adsorption capacity (NAC), glucose adsorption capacity (GAC), glucose dialysis retardation index (GDRI), α-amylase inhibitory activity, and DPPH radical scavenging activity, were far better than those of ODF. Together, the results revealed that AHP and EM modifications could effectively improve or enhance the physicochemical and functional properties of Forsythia suspensa DF. [ABSTRACT FROM AUTHOR]

Forsythia suspensa is a deciduous shrub that belongs to the family Myrtaceae, and its dried fruits are used as medicine. F. suspensa contains several secondary metabolites, which exert pharmacological effects. One of the main active components is forsythin, which exhibits free radical scavenging, antioxidant, anti-inflammatory, and anti-cancer effects. Mitogen-activated protein kinase (MAPKs) can increase the activity of WRKY family transcription factors in a phosphorylated manner, thereby increasing the content of secondary metabolites. However, the mechanism of interaction between MAPKs and WRKYs in F. suspensa remains unclear. In this study, we cloned the genes of FsWRKY4 and FsMAPK3, and performed a bioinformatics analysis. The expression patterns of FsWRKY4 and FsMAPK3 were analyzed in the different developmental stages of leaf and fruit from F. suspensa using real-time fluorescence quantitative PCR (qRT-PCR). Subcellular localization analysis of FsWRKY4 and FsMAPK3 proteins was performed using a laser scanning confocal microscope. The existence of interactions between FsWRKY4 and FsMPAK3 in vitro was verified by yeast two-hybridization. Results showed that the cDNA of FsWRKY4 (GenBank number: OR566682) and FsMAPK3 (GenBank number: OR566683) were 1587 and 522 bp, respectively. The expression of FsWRKY4 was higher in the leaves than in fruits, and the expression of FsMAPK3 was higher in fruits but lower in leaves. The subcellular localization results indicated that FsWRKY4 was localized in the nucleus and FsMAPK3 in the cytoplasm and nucleus. The prey vector pGADT7-FsWRKY4 and bait vector pGBKT7-FsMAPK3 were constructed and co-transferred into Y2H Glod yeast receptor cells. The results indicated that FsWRKY4 and FsMAPK3 proteins interact with each other in vitro. The preliminary study may provide a basis for more precise elucidation of the synthesis of secondary metabolites in F. suspensa. [ABSTRACT FROM AUTHOR]

Weeping forsythia is an important ornamental, ecological and medicinal plant. Brown leaf spots limit the large-scale production of weeping forsythia as a medicinal crop. Alternaria alternata is a pathogen causing brown leaf spots in weeping forsythia; however, its pathogenesis and the immune response mechanisms of weeping forsythia remain unclear. In this study, we identified two mechanisms based on morphological anatomy, physiological indexes and gene expression analyses. Our results showed that A. alternata induced leaf stomata to open, invaded the mesophyll, dissolved the cell wall, destroyed the cell membrane and decreased the number of chloroplasts by up-regulating the expression of auxin-activated signaling pathway genes. Alternaria alternata also down-regulated iron-ion homeostasis and binding-related genes, which caused an increase in the levels of iron ions and reactive oxygen species in leaves. These processes eventually led to programmed cell death, destroying palisade and spongy tissues and causing the formation of iron rust spots. Alternaria alternata also caused defense and hypersensitive responses in weeping forsythia through signaling pathways mediated by flg22-like and elf18-like polypeptides, ethylene, H2O2 and bacterial secretion systems. Our study provides a theoretical basis for the control of brown leaf spots in weeping forsythia. [ABSTRACT FROM AUTHOR]

In this experiment, Forsythia suspensa was used as raw material to extract insoluble dietary fiber (IDF) by double enzyme method, and then the extracted IDF was chemically modified by carboxymethylation (CM), hydroxypropylation (HP) and cross-linking (CL), and their physicochemical properties, structure and emulsifying properties were analyzed. The results showed that the modified treatment reduced the water holding capacity of Forsythia suspensa IDF and significantly increased the oil holding capacity by 21%~53.0% (P

In this study, the process of steam distillation to collect volatile oils from Forsythia suspensa (F. suspensa) and Lonicera japonica (L. japonica) was optimized according to the concept of quality by design. First, the liquid/material ratio, distillation time, and collection temperature were identified as critical process parameters by a review of the literature and single-factor experiments. Then, a Box–Behnken design was used to study the quantitative relationship between the three process parameters, two raw material properties, and the yield of volatile oil. A mathematical model was established with an R2 value exceeding 0.90. Furthermore, the design space of the volatile oil yield was calculated by a probability-based method. The results of a verification experiment showed that the model was accurate and the design space was reliable. A total of 16 chemical constituents were identified in the volatile oil from mixtures of F. suspensa and L. japonica. The content of β-pinene was the highest (54.75%), and the composition was similar to that of the volatile oil of F. suspensa. The results showed that when F. suspensa and L. japonica were distilled together, the main contribution to the volatile oil was from F. suspensa. The volatile oil yield from the combination of F. suspensa and L. japonica was not higher than that from L. japonica. [ABSTRACT FROM AUTHOR]

Besides active substances, Forsythia suspensa is rich in dietary fiber (DF), but it is often wasted or discarded and not put to good use. In order to improve the function of Forsythia DF, it was modified using alkaline hydrogen peroxide (AHP) and cellulase (EM). Compared to the control DF (ODF), the DF modified using AHP (AHDF) and EM (EMDF) had a looser microstructure, lower crystallinity, and higher oil holding capacity (OHC) and cation exchange capacity (CEC). The AHP treatment significantly increased the water holding capacity (WHC) and water swelling ability (WSA) of the DF, while the EM treatment achieved just the opposite. Moreover, the functional properties of AHDF and EMDF, including their cholesterol adsorption capacity (CAC), nitrite ion adsorption capacity (NAC), glucose adsorption capacity (GAC), glucose dialysis retardation index (GDRI), α-amylase inhibitory activity, and DPPH radical scavenging activity, were far better than those of ODF. Together, the results revealed that AHP and EM modifications could effectively improve or enhance the physicochemical and functional properties of Forsythia suspensa DF.

Forsythia suspensa is a deciduous shrub that belongs to the family Myrtaceae, and its dried fruits are used as medicine. F. suspensa contains several secondary metabolites, which exert pharmacological effects. One of the main active components is forsythin, which exhibits free radical scavenging, antioxidant, anti-inflammatory, and anti-cancer effects. Mitogen-activated protein kinase (MAPKs) can increase the activity of WRKY family transcription factors in a phosphorylated manner, thereby increasing the content of secondary metabolites. However, the mechanism of interaction between MAPKs and WRKYs in F. suspensa remains unclear. In this study, we cloned the genes of FsWRKY4 and FsMAPK3, and performed a bioinformatics analysis. The expression patterns of FsWRKY4 and FsMAPK3 were analyzed in the different developmental stages of leaf and fruit from F. suspensa using real-time fluorescence quantitative PCR (qRT-PCR). Subcellular localization analysis of FsWRKY4 and FsMAPK3 proteins was performed using a laser scanning confocal microscope. The existence of interactions between FsWRKY4 and FsMPAK3 in vitro was verified by yeast two-hybridization. Results showed that the cDNA of FsWRKY4 (GenBank number: OR566682) and FsMAPK3 (GenBank number: OR566683) were 1587 and 522 bp, respectively. The expression of FsWRKY4 was higher in the leaves than in fruits, and the expression of FsMAPK3 was higher in fruits but lower in leaves. The subcellular localization results indicated that FsWRKY4 was localized in the nucleus and FsMAPK3 in the cytoplasm and nucleus. The prey vector pGADT7-FsWRKY4 and bait vector pGBKT7-FsMAPK3 were constructed and co-transferred into Y2H Glod yeast receptor cells. The results indicated that FsWRKY4 and FsMAPK3 proteins interact with each other in vitro. The preliminary study may provide a basis for more precise elucidation of the synthesis of secondary metabolites in F. suspensa.

The effects of varying concentrations of Bacillus subtilis suspensions on the growth physiology and Cd accumulation of Forsythia suspense under moderate Cd pollution stress were studied using a pot experiment. The results showed that Bacillus subtilis suspension significantly alleviated Cd stress on the growth of F. suspense (P＜0.05). Compared with those in the Cd treatment group (15 mg·kg-1 Cd+200 mL tap water), the plant height, fresh weight of stems and leaves, fresh weight of roots, chlorophyll a content in leaves, and total chlorophyll content in the H treatment group (15 mg·kg-1 Cd+200 mL bacteria solution) increased by 112.59%, 96.50%, 95.68%, 210.53%, and 162.30%, respectively. Different concentrations of bacterial suspensions significantly decreased Cd content in the roots, stems, leaves, and rhizosphere soil of F. suspensa; increased superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activity; and decreased malondialdehyde (MDA) content in the leaves of F. suspensa (P＜0.05). Compared with those in the Cd treatment group, the Cd content in the roots, stems, leaves, and rhizosphere soil of F. suspense seedlings treated with different concentrations of bacterial suspension decreased by 34.23%, 29.35%, and 44.01%, respectively; the activities of SOD, POD, and CAT increased by 530.48%, 103.78%, and 133.33%, respectively; and the MDA content in the leaves reduced by 64.41%. These results showed that Bacillus subtilis suspension could significantly alleviate Cd stress on the growth of F. suspensa. By increasing the activities of SOD, POD, and CAT in leaves and reducing lipid peroxidation damage, the tolerance of plants to Cd improved, and the accumulation of Cd in F. suspensa was inhibited. [ABSTRACT FROM AUTHOR]

Forsythia suspensa (Thunb.) has been widely used in traditional medicines in Asia. According to the 2020 edition of Chinese Pharmacopoeia, phillyrin is the main active ingredient in F. suspensa, which is effective in clearing heat, reducing swelling, and dispersing nodules. F. suspensa leaf is a non-toxic substance and it can be used to make a health tea. Here, we combine elicitors and transcriptomics to investigate the inducible biosynthesis of the phillyrin from the F. suspensa. After the fruits and leaves of F. suspensa were treated with different concentrations of methyl jasmonate (MeJA), the content of phillyrin in the fruits reached a peak at 200 µM MeJA for 12 h, but which was decreased in leaves. To analyze the differences in key enzyme genes involved in the phillyrin biosynthesis, we sequenced the transcriptome of F. suspensa leaves and fruits treated with 200 µM MeJA for 12 h. We hypothesized that nine genes related to coniferin synthesis including: F. suspensa UDP-glycosyltransferase (FsUGT); F. suspensa 4-coumarate coenzyme CoA ligase (Fs4CL); and F. suspensa Caffeoyl-CoA O-methyltransferase (FsCCoAOMT) etc. The qRT-PCR analysis of genes related to phillyrin biosynthesis was consistent with RNA-seq analysis. We also investigated the dynamic changes of genes in F. suspensa leaves and fruits at different time points after 200 µM MeJA treatment, which laid the foundation for further study of the molecular mechanisms regulating the biosynthesis of phillyrin. [ABSTRACT FROM AUTHOR]

Forsythia suspensa is a famous ornamental and medicinal plant in Oleaceae. CCD family is involved in the synthesis of pigments, volatiles, strigolactones, and abscisic acid (ABA) in plants. In this study, the CCD family in F. suspensa was analyzed at the genome level. A total of 16 members of the CCD family were identified, which included 11 members of the carotenoid cleavage dioxygenases (CCD) subfamily and 5 members of the 9-cis epoxycarotenoid dioxygenases (NCED) subfamily. The expression analysis of different tissues demonstrated that three FsCCD1 genes might be involved in the synthesis of pigments and volatiles in flowers and fruits. Three CCD4 genes were effectively expressed in flowers, while only FsCCD4-3 was effectively expressed in fruits. Comparison of CCD4 between Osmanthus fragrans and F. suspensa showed that the structure of FsCCD4-1 is was comparable that of OfCCD4-1 protein, indicating that the protein might be performing, especially in catalyzing the synthesis of b-ionone. However, further comparison of the upstream promoter regions showed that the proteins have major differences in the composition of cis-elements, which might be responsible for differences in b-ionone content. On the other hand, four NCED genes were significantly up-regulated under cold stress while two were up-regulated in drought stress. The data showed that these genes might be involved in the synthesis of ABA. Taken together, our data improves understanding of the CCD family and provides key candidate genes associated with cold and drought stresses in F. suspensa. [ABSTRACT FROM AUTHOR]

Forsythiasides are a kind of phenylethanol glycosides existing in Forsythia suspensa (Thunb.) Vahl, which possesses extensive pharmacological activities. According to the different groups connected to the nucleus, forsythiasides can be divided into A-K. In recent years, numerous investigations have been carried out on forsythiasides A, B, C, D, E, and I, which have the effects of cardiovascular protection, anti-inflammation, anti-oxidation, neuroprotection, et al. Mechanistically, forsythiasides regulate toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor kappaB (NF-κB), nuclear factor-erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) and other signaling pathways, as well as the expression of related cytokines and kinases. Further exploration and development may unearth more treatment potential of forsythiasides and provide more evidence for their clinical applications. In summary, forsythiasides have high development and application value.

• The enzyme and ultrasonic treatment accelerate cell rupture and release the active ingredients. • FS was extracted by ultrasound-assisted β-cyclodextrin in water. • Different extraction processes were evaluated in terms of energy consumption, CO 2 production. • EUA-β-CDE increased the antioxidant and anti-inflammatory activities of FS. With the proposal of the 2030 Agenda for Sustainable Development, the Chinese medicine extraction technology has been innovatively improved to prioritize low energy consumption, sustainability, and minimized organic solvent utilization. Forsythia suspensa (FS) possesses favorable pharmacological properties and is extensively utilized in traditional Chinese medicine. However, due to the limitations of the composition and extraction methods, its potential has not been fully developed. Thus, a combination of ultrasound-assisted extraction (UAE), enzyme-assisted extraction (EAE), and β-cyclodextrin extraction (β-CDE) was employed to isolate and purify rutin, phillyrin, and forsythoside A from FS. The results demonstrated that the efficiency of extracting enzymatic and ultrasound assisted β-cyclodextrin extraction (EUA-β-CDE) was highly influenced by the temperature and duration of hydrolysis, as well as the duration of the extraction process. According to the results of the single-factor experiment, Box-Behnken design (BBD) in Response surface method (RSM) was used to optimize the experimental parameters to achieve the maximum comprehensive evaluation value (CEV) value. The EUA-β-CDE compared with other extraction methods, has good extraction effect and low energy consumption by high performance liquid chromatography (HPLC), scanning electron microscopy (SEM), calculation of power consumption and CO 2 emission The EUA-β-CDE compared with other extraction methods, has good extraction effect and low energy consumption by HPLC, SEM, calculation of power consumption and CO 2 emission. Then, the structural characteristics of EUA-β-CDE of FS extract had significant interaction with β-CD by Fourier infrared spectroscopy (FT-IR) and differential scanning calorimetry (DSC). In addition, EUA-β-CDE extract has good antioxidant and anti-inflammatory activities. The establishment of EUA-β-CDE of FS provides a new idea for the development and application of other sustainable extraction methods of traditional Chinese medicine. [ABSTRACT FROM AUTHOR]

With the widespread use and abuse of antibiotics, Helicobacter pylori (H. pylori) has become seriously drug resistant. The development of new antibiotics is an important way to solve H. pylori 's drug resistance. Screening antibacterial ingredients from natural products is a convenient way to develop new antibiotics. Phillygenin, an effective antibacterial component, was selected from the natural product, forsythia, in this study. Its minimal inhibitory concentration (MIC) for 18 H. pylori strains was 16–32 μg/ml. The minimum bactericidal concentration (MBC) of H. pylori G27 was 128 μg/ml; the higher the drug concentration and the longer the time, the better the sterilization effect. It was non-toxic to gastric epithelial cell (GES)-1 and BGC823 cells at the concentration of 100 μg/ml. It presented a better antibacterial effect on H. pylori in an acidic environment, and after 24 days of induction on H. pylori with 1/4 MIC of phillygenin, no change was found in the MIC of H. pylori. In the mechanism of action, phillygenin could cause ATP leakage and inhibit the biofilm formation; the latter was associated with the regulation of spoT and Hp1174 genes. In addition, phillygenin could regulate the genes of Nhac, caggamma, MATE, MdoB, flagellinA , and lptB , leading to the weakening of H. pylori 's acid resistance and virulence, the diminishing of H. pylori 's capacity for drug efflux, H. pylori 's DNA methylation, the initiation of human immune response, and the ATP leakage of H. pylori , thus accelerating the death of H. pylori. In conclusion, phillygenin was a main ingredient inhibiting H. pylori in Forsythia suspensa , with a good antibacterial activity, high safety, strong specificity, better antibacterial effect under acidic conditions, and low risk of resistance development by H. pylori. Its mechanism of action was mainly associated with inhibiting the biofilm formation and resulting in ATP leakage. In addition, phillygenin was shown to be able to reduce the acid resistance and virulence of H. pylori. [ABSTRACT FROM AUTHOR]

With the widespread use and abuse of antibiotics, Helicobacter pylori (H. pylori) has become seriously drug resistant. The development of new antibiotics is an important way to solve H. pylori's drug resistance. Screening antibacterial ingredients from natural products is a convenient way to develop new antibiotics. Phillygenin, an effective antibacterial component, was selected from the natural product, forsythia, in this study. Its minimal inhibitory concentration (MIC) for 18 H. pylori strains was 16–32 μg/ml. The minimum bactericidal concentration (MBC) of H. pylori G27 was 128 μg/ml; the higher the drug concentration and the longer the time, the better the sterilization effect. It was non-toxic to gastric epithelial cell (GES)-1 and BGC823 cells at the concentration of 100 μg/ml. It presented a better antibacterial effect on H. pylori in an acidic environment, and after 24 days of induction on H. pylori with 1/4 MIC of phillygenin, no change was found in the MIC of H. pylori. In the mechanism of action, phillygenin could cause ATP leakage and inhibit the biofilm formation; the latter was associated with the regulation of spoT and Hp1174 genes. In addition, phillygenin could regulate the genes of Nhac, caggamma, MATE, MdoB, flagellinA, and lptB, leading to the weakening of H. pylori's acid resistance and virulence, the diminishing of H. pylori's capacity for drug efflux, H. pylori's DNA methylation, the initiation of human immune response, and the ATP leakage of H. pylori, thus accelerating the death of H. pylori. In conclusion, phillygenin was a main ingredient inhibiting H. pylori in Forsythia suspensa, with a good antibacterial activity, high safety, strong specificity, better antibacterial effect under acidic conditions, and low risk of resistance development by H. pylori. Its mechanism of action was mainly associated with inhibiting the biofilm formation and resulting in ATP leakage. In addition, phillygenin was shown to be able to reduce the acid resistance and virulence of H. pylori.

In this study, the process of steam distillation to collect volatile oils from Forsythia suspensa (F. suspensa) and Lonicera japonica (L. japonica) was optimized according to the concept of quality by design. First, the liquid/material ratio, distillation time, and collection temperature were identified as critical process parameters by a review of the literature and single-factor experiments. Then, a Box–Behnken design was used to study the quantitative relationship between the three process parameters, two raw material properties, and the yield of volatile oil. A mathematical model was established with an R2 value exceeding 0.90. Furthermore, the design space of the volatile oil yield was calculated by a probability-based method. The results of a verification experiment showed that the model was accurate and the design space was reliable. A total of 16 chemical constituents were identified in the volatile oil from mixtures of F. suspensa and L. japonica. The content of β-pinene was the highest (54.75%), and the composition was similar to that of the volatile oil of F. suspensa. The results showed that when F. suspensa and L. japonica were distilled together, the main contribution to the volatile oil was from F. suspensa. The volatile oil yield from the combination of F. suspensa and L. japonica was not higher than that from L. japonica.

Forsythia suspensa (Thunb.) has been widely used in traditional medicines in Asia. According to the 2020 edition of Chinese Pharmacopoeia, phillyrin is the main active ingredient in F. suspensa, which is effective in clearing heat, reducing swelling, and dispersing nodules. F. suspensa leaf is a non-toxic substance and it can be used to make a health tea. Here, we combine elicitors and transcriptomics to investigate the inducible biosynthesis of the phillyrin from the F. suspensa. After the fruits and leaves of F. suspensa were treated with different concentrations of methyl jasmonate (MeJA), the content of phillyrin in the fruits reached a peak at 200 µM MeJA for 12 h, but which was decreased in leaves. To analyze the differences in key enzyme genes involved in the phillyrin biosynthesis, we sequenced the transcriptome of F. suspensa leaves and fruits treated with 200 µM MeJA for 12 h. We hypothesized that nine genes related to coniferin synthesis including: F. suspensa UDP-glycosyltransferase (FsUGT); F. suspensa 4-coumarate coenzyme CoA ligase (Fs4CL); and F. suspensa Caffeoyl-CoA O-methyltransferase (FsCCoAOMT) etc. The qRT-PCR analysis of genes related to phillyrin biosynthesis was consistent with RNA-seq analysis. We also investigated the dynamic changes of genes in F. suspensa leaves and fruits at different time points after 200 µM MeJA treatment, which laid the foundation for further study of the molecular mechanisms regulating the biosynthesis of phillyrin.

Forsythia suspensa (Thunb.) Vahl (FS) is an important plant with high edible and medicinal values. The edible fruit of FS is a common traditional medicine in China, Japan and Korea. Compared to the research on the phytochemistry and pharmacology of the fruits, study on the other parts of FS , such as leaves and flowers is still limited. In this study, an integrated strategy based on ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF-MS/MS), plant metabolomics and correlation analysis was established for comprehensively chemical characterization of fruits, leaves and flowers of FS, and discovery of α-glucosidase and pancreatic lipase inhibitory metabolites. The plant metabolic profiling of fruits, leaves and flowers of FS was performed by UPLC-QTOF-MS/MS, and a total of 74 secondary metabolites, including 15 phenylethanoid glycosides, 20 lignans, 10 cyclohexanol derivatives, 11 organic acids, 9 flavonoids, 3 triterpenes, and 10 other compounds were identified. Then, 29 differential secondary metabolites that responsible to distinguish the fruits, leaves and flowers of FS were further screened out by multivariate statistical analysis. Meanwhile, the α-glucosidase and pancreatic lipase inhibition of different parts of FS were evaluated and compared by in vitro experiments. The results demonstrated that the leaves of FS showed the highest inhibition on α-glucosidase and pancreatic lipase with IC 50 of 0.17 ± 0.04 mg/mL and 0.56 ± 0.33 mg/mL, respectively. Then, the correlation between differential metabolites and enzyme inhibitory activities were investigated by Pearson correlation analysis, and 12 potential α-glucosidase and pancreatic lipase inhibitors were screen out. Additionally, the α-glucosidase and pancreatic lipase inhibitory activities of five potential enzyme inhibitory components, including quercitrin, rutin, kaempferol-3-O-rutinoside, pinoresinol-4-O-glucoside and phillyrin were further validated by in vitro assays and molecular docking analysis. The results showed that all the five potential inhibitors showed good inhibitory effects on α-glucosidase and pancreatic lipase, and the binding of the five inhibitors to enzymes mainly through hydrogen bonding, hydrophobic force, and ionic bonding. This study provided a feasible strategy for comparison and discrimination of different parts of medicinal plant and discovery of bioactive components, and also provided useful information for future utilization of different parts of FS. [ABSTRACT FROM AUTHOR]

Abstract Background The dried fruits of Forsythia suspensa has generally been used to clear heat and detoxify in traditional Korean and Chinese medicine. Oxaliplatin is a first-line treatment chemotherapeutic agent for advanced colorectal cancer, but it induces peripheral neuropathy as an adverse side effect affecting the treatment regimen and the patient’s quality of life. The present study was conducted to evaluate the neuroprotective effects of an aqueous extract of F. suspensa fruits (EFSF) on oxaliplatin-induced peripheral neuropathy. Methods The chemical components from EFSF were characterized and quantified using the ultra-high performance liquid chromatography-diode array detector system. The cytotoxicities of anticancer drugs in cancer cells and PC12 cells were assessed by the Ez-Cytox viability assay. To measure the in vitro neurotoxicity, the neurite outgrowth was analyzed in the primary dorsal root ganglion (DRG) cells, and neural PC12 cells that were differentiated with nerve growth factor. To evaluate the in vivo neuroprotective activity, the von Frey test was performed in six-week-old male mice (C57BL/6) receiving EFSF (60–600 mg/kg) in the presence of 20–30 mg/kg cumulative doses of oxaliplatin. Thereafter, the mice were euthanized for immunohistochemical staining analysis with an antibody against PGP9.5. Results EFSF attenuated the cytotoxic activities of the various anticancer drugs in neural PC12 cells, but did not affect the anticancer activity of oxaliplatin in human cancer cells. Oxaliplatin remarkably induced neurotoxicities including cytotoxicity and the inhibited neurite outgrowth of DRG and neural PC12 cells. However, the co-treatment of EFSF (100 μg/ml) with oxaliplatin completely reversed the oxaliplatin-induced neurotoxicity. Forsythoside A, the major component of EFSF, also exerted remarkable neuroprotective effects against the oxaliplatin-induced neurotoxicity. In addition, EFSF (60–200 mg/kg) significantly alleviated the oxaliplatin-induced mechanical allodynia and loss of intra-epidermal nerve fiber to the levels of the vehicle control in the mouse peripheral neuropathy model. Conclusions EFSF could be considered a useful herbal medicine for the treatment of peripheral neuropathy in cancer patients receiving chemotherapy with oxaliplatin.

Objective: To compare the anti-inflammatory, antitumor and anti-bacterial effects of the single extract (in granules) and the prepared drug in pieces of Forsythia Suspense ( Lianqiao , a traditional Chinese herbal medicine)., Methods: In zebrafish embryo models of CuSO 4 exposure, tail transection and LPS microinjection-induced inflammation, the anti-inflammatory effects of 10 μg/mL DEX, single extract of Forsythia Suspense, and the water extract of the prepared drug (400, 600, and 800 μg/mL) were evaluated by observing neutrophil counts, RT- qPCR, HE staining and survival analysis. Zebrafish embryo models bearing different human tumor cell xenografts were used to assess the anti-tumor effect of the drugs in different dosage forms by fluorescence staining and HE staining. The microbroth dilution method was used to evaluate the antibacterial efficacy of the drugs., Results: In the zebrafish embryo models of inflammation, both of the two dosage forms of Forsythia Suspense significantly inhibited neutrophil aggregation, reduced the mRNA expressions of TNF-α, IL-6, P38, Jnk, Erk and P65, and increased the survival rate of zebrafish. They both showed obvious inhibitory effects against xenografts of different human cancer cells including colon cancer cells (HCT116), pancreas adenocarcinoma cells (PANC-1), lung cancer cells (A549), liver cancer cells (Hep3B) and cervical carcinoma cells (Hela) in zebrafish embryos, and exhibited strong anti-bacterial effects at the concentration of 15.63 mg/mL., Conclusion: The two dosage forms of Forsythia Suspense have similar anti-inflammatory, antitumor and antibacterial effects, but their effects for inhibiting IL-6, P65, and Jnk mRNA expressions and HCT116 cell proliferation differ significantly at low doses in zebrafish.

This study systematically evaluated the effect of Forsythia suspensa extract on dextran sodium sulfate (DSS)-induced ulcerative colitis (UC) and determined its mechanism of action. The results showed that Forsythia suspensa extract significantly inhibited DSS-induced UC in mice. In vivo mechanistic studies revealed that Forsythia suspensa extract relieved the symptoms of colitis by enhancing antioxidant activity and inhibiting pyroptosis. Further in vitro experiments on the mechanism of Forsythia suspensa showed that it reduced the level of reactive oxygen species (ROS) in J774A.1 cells. We found that Forsythia suspensa extract enhanced cellular antioxidation activity and inhibited pyroptosis. After silencing NLRP3, it was found to play an important role in pyroptosis. In addition, after Nrf2 was silenced, the inhibitory effect of Forsythia suspensa extract on cell pyroptosis was eliminated, indicating an interaction between Nrf2 and NLRP3. Metabonomics revealed that Forsythia suspensa extract significantly improved metabolic function in colitis mice by reversing the abnormal changes in the levels of 9 metabolites. The main metabolic pathways involved were glutathione metabolism, aminoacyl-tRNA biosynthesis and linoleic acid metabolism. In conclusion, we found that Forsythia suspensa extract significantly alleviated DSS-induced UC injury through the Nrf2-NLRP3 pathway and relieved metabolic dysfunction.

Forsythia suspensa is an important medicinal plant and native to China. In this article, the complete chloroplast genome of F. suspensa was studied and annotated. The chloroplast genome of F. suspensa is 156,404 bp in length, which has a large single-copy region (LSC) of 87,150 bp, a small single-copy region (SSC) of 17,812 bp, and two of inverted-repeat regions (IRs) of 25,716 bp. The overall nucleotide composition is: 30.8% (48,103 bp) of A, 31.4% (49,193 bp) of T, 19.2% (30,066 bp) of C and 18.6% (29,042 bp) of G, and the total content of the chloroplast genome A + T content of 62.2% and G + C content of 37.8%. The chloroplast genome of F. suspensa contains 133 genes, including 88 protein-coding genes (PCGs), 37 transfer RNA (tRNAs), and 8 ribosome RNA (rRNAs). The phylogenetic tree result shows that F. suspensa is closely related to F. intermedia in the phylogenetic relationship by the maximum-likelihood (ML) method.

Purpose: To investigate the effect of Forsythia suspensa extract (FSE) on apoptosis and proliferation in A549 human lung cancer cells. Methods: Inverted microscope was employed to observe morphological changes in A549 cells after exposure to FSE. Trypan blue staining of living cells was used to construct the cell growth curve after treatment with varying concentrations of FSE. The influence of FSE on cell proliferation, apoptosis and cell cycle was determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay, while protein expressions of key apoptosis-related enzymes were evaluated by immunocytochemical method. Results: FSE inhibited the growth of A549 lung cancer cells at a concentration range of 10 - 150 µg/mL. Flow cytometry results showed that FSE induced apoptosis in A549 cells. The proportion of cells in G0/G1-phase increased significantly (p < 0.01), while the proportion of cells in S- and G2/M-phase decreased correspondingly, indicating that the cells were in G0/G1-phase arrest. Cell cycle arrest and apoptosis-inducing effect gradually rose with increase in FSE concentration. With increasing concentrations of FSE, there was also significant increase in the expressions of caspase-3 (p < 0.05), caspase-8 (p < 0.01) and caspase-9 (p < 0.05), but significant decrease in Ki-67 (p < 0.01) and p21 ras protein (p < 0.01). Conclusion: FSE exerts significant inhibitory effect on the proliferation of A549 lung cancer cells. Therefore, the plant can potentially be developed for the treatment of lung cancer. [ABSTRACT FROM AUTHOR]

Forsythiae Fructus (FF), the dried fruit of Forsythia suspensa, has been widely used as a heat-clearing and detoxifying herbal medicine in China. Green FF (GF) and ripe FF (RF) are fruits of Forsythia suspensa at different maturity stages collected about a month apart. FF undergoes a complex series of physical and biochemical changes during fruit ripening. However, the clinical uses of GF and RF have not been distinguished to date. In order to comprehensively compare the chemical compositions of GF and RF, NMR-based metabolomics coupled with HPLC and UV spectrophotometry methods were adopted in this study. Furthermore, the in vitro antioxidant and antibacterial activities of 50% methanol extracts of GF and RF were also evaluated. A total of 27 metabolites were identified based on NMR data, and eight of them were found to be different between the GF and RF groups. The GF group contained higher levels of forsythoside A, forsythoside C, cornoside, rutin, phillyrin and gallic acid and lower levels of rengyol and β-glucose compared with the RF group. The antioxidant activity of GF was higher than that of RF, but no significant difference was observed between the antibacterial activities of GF and RF. Given our results showing their distinct chemical compositions, we propose that NMR-based metabolic profiling can be used to discriminate between GF and RF. Differences in the chemical and biological activities of GF and RF, as well as their clinical efficacies in traditional Chinese medicine should be systematically investigated in future studies.

An efficient ultrasonic-assisted extraction (UAE) method was developed for simultaneous extraction of three active compounds, forsythiaside A (FSA), phillyrin (PHI) and rutin (RT), from the fruits of Forsythia suspensa. The effects of various factors including a binary mixed solvent of methanol/water and ethanol/water, the pH of the solvent, particle size, temperature, solvent to material ratio, ultrasonic input power and extraction time on UAE were investigated in detail. The mass transfer mechanism of UAE using different mixed solvents was further explained by comparison with the maceration extraction method. The response surface methodology was used to optimize the experimental variables including ethanol concentration, solvent to material ratio and extraction time. The optimized conditions for the simultaneous extraction of RT, FSA and PHI were: particle size 60-80 mesh, temperature 30 °C, ultrasonic power 200 W, ethanol concentration 50%, solvent to material ratio 32 mL/g and extraction time 37 min. Compared to conventional extraction methods, UAE provided the highest extraction efficiency and offered many advantages including the reduction of solvent, temperature and time for extraction. [ABSTRACT FROM AUTHOR]

The aim of the study was to develop an alternative capillary zone electrophoresis (CZE) for simultaneous determination of phillyrin (1), phillygenin (2), epipinoresinol-4-O-β-glucoside (3), pinoresinol-4-O-β-glucoside (4), lariciresinol (5), pinoresinol (6), isolariciresinol (7) and vladinol D (8) in Forsythia suspensa. The structural types of lignans 1-8 could be attributed to bisepoxylignans (1-4 and 6), monoepoxylignans (5 and 8) and cyclolignan (7). The major difficulties in the CZE separation of 1-8 could be summarization as the simultaneous presence of free lignans (1, 2 and 5-8) and lignan glucosides (3 and 4) and simultaneous occurrence of two pairs of isomers (3 and 4 as well as 5 and 7). Without the addition of β-CD and methanol, the resolution of these analytes was quite poor. However, in this study, compounds 1-8 were excellently separated from each other within 15 min under optimized conditions with a borax running buffer (40 mM borax, pH 10.30) containing 2 mM β-CD and 5% methanol (v/v) at the voltage of 20 kV, temperature of 35 °C and detection wavelength of 234 nm. Validation of the method included tests of linearity, precision, repeatability, stability and accuracy. In addition, the method offered inherent advantages such as lower analytical cost, no need of specific columns and use of small amounts of organic solvents and reagents. Finally, this green and economic CZE was successfully applied for the determination of these bioactive components 1-8 in F. suspensa fruits and its commercial extracts. [ABSTRACT FROM AUTHOR]

A novel iridoid glycoside trimer named forsydoitriside A ( 1 ) and five new iridoid glycosides ( 2–6 ) were isolated from the fruits of Forsythia suspensa together with two known compounds ( 7 , 8 ). These new structures were elucidated by comprehensive spectroscopic data and the comparison of experimental and calculated electronic circular dichroism spectra. Compounds 1 – 8 were all assayed on acetaminophen-induced HepG2 cell damage. The results exhibited that compounds 2 , 3 , 5 and 6 possessed strong hepatoprotective activities against the damage in HepG2 cell. [ABSTRACT FROM AUTHOR]

Generally, essential oils and components of interest are extracted from plants using organic solvent, distillation, ultrasound and supercritical extraction methods. Ultrasonic extraction (UE) has the advantage of high efficiency, but its process is complicated and it has numerous variables. In this study, an L18-Hunter experimental design was applied for the first time to investigate the practicability of applying UE to Forsythia suspensa seed oil. Six potential high-risk variables, including numerical and non-numeric types, were obtained from the risk analysis and their impacts on global yield and antioxidant activity were screened. Furthermore, oils obtained by different extraction processes (i.e., UE, supercritical fluid extraction (SFE), soxhlet extraction (SE) and hydrodistillation extraction (HD)) were analyzed. A comparative study of these oils was characterized and compared by FT-IR, GC-MS and antioxidant activity. The obtained results show that the type of solvent, solvent-to-solid ratio, extraction power and time were the significant variables affecting the extraction yield, whereas antioxidant activity was only affected by the type of solvent. The regression coefficients of the yield and antioxidant activity models were 0.79 and 0.91, and the ANOVA of the models were 0.013 and Forsythia suspensa seed oil extraction process, but also provided a time- and cost-effective advantage to the food and pharmaceutical industry when compared to the SFE, SE and HD extraction processes.

Forsythia suspensa is an important medicinal plant and traditionally applied for the treatment of inflammation, pyrexia, gonorrhea, diabetes, and so on. However, there is limited sequence and genomic information available for F. suspensa. Here, we produced the complete chloroplast genomes of F. suspensa using Illumina sequencing technology. F. suspensa is the first sequenced member within the genus Forsythia (Oleaceae). The gene order and organization of the chloroplast genome of F. suspensa are similar to other Oleaceae chloroplast genomes. The F. suspensa chloroplast genome is 156,404 bp in length, exhibits a conserved quadripartite structure with a large single-copy (LSC; 87,159 bp) region, and a small single-copy (SSC; 17,811 bp) region interspersed between inverted repeat (IRa/b; 25,717 bp) regions. A total of 114 unique genes were annotated, including 80 protein-coding genes, 30 tRNA, and four rRNA. The low GC content (37.8%) and codon usage bias for A- or T-ending codons may largely affect gene codon usage. Sequence analysis identified a total of 26 forward repeats, 23 palindrome repeats with lengths >30 bp (identity >90%), and 54 simple sequence repeats (SSRs) with an average rate of 0.35 SSRs/kb. We predicted 52 RNA editing sites in the chloroplast of F. suspensa, all for C-to-U transitions. IR expansion or contraction and the divergent regions were analyzed among several species including the reported F. suspensa in this study. Phylogenetic analysis based on whole-plastome revealed that F. suspensa, as a member of the Oleaceae family, diverged relatively early from Lamiales. This study will contribute to strengthening medicinal resource conservation, molecular phylogenetic, and genetic engineering research investigations of this species. [ABSTRACT FROM AUTHOR]

Understanding the genetic mechanisms of adaptation to environmental variables is a key concern in molecular ecology and evolutionary biology. Determining the adaptive evolutionary direction and evaluating the adaptation status of species can improve our understanding of these mechanisms. In this study, we sampled 20 populations of Forsythia suspensa to infer the relationship between environmental variables and adaptive genetic variations. Population structure analysis revealed that four genetic groups of F. suspensa exist resulting from divergent selection driven by seven environmental variables. A total of 26 outlier loci were identified by both BayeScan and FDIST2, 23 of which were environment-associated loci (EAL). Environmental association analysis revealed that the environmental variables related to the ecological habitats of F. suspensa are associated with high numbers of EAL. Results of EAL characterization in F. suspensa are consistent with the hypothesis that ecological habitats determine the adaptive evolution of this species. Moreover, a model of species adaptation to environmental variables was proposed in this study. The adaptation model was used to further evaluate the adaptation status of F. suspensa to environmental variables. This study will be useful to help us in understanding the adaptive evolution of species in regions lacking strong selection pressure. [ABSTRACT FROM AUTHOR]

Chemical investigation of the 70% ethanol extract of the unripe fruits of Forsythia suspensa resulted in the isolation of a novel caffeoyl phenylethanoid glycoside, lianqiaoxinoside B, together with the known compound forsythoside H. The new compound was elucidated to be 1'',2''-[β-(3,4,-dihydroxylphenyl)-α,β-dioxoethanol]-3''-O-caffeoyl-O-α-rhamnopyranosyl-(1→6)-O-β-glucopyranoside by extensive spectroscopic and chemical studies. Lianqiaoxinoside B and forsythoside H showed strong antioxidant and antimicrobial activities in vitro by the 2,2'-azinobis-3-ethylbenzothiazoline-6-sulphonate (ABTS) radical-scavenging assay and plate method. This study can be further extended to exploit for the possible application of caffeoyl phenylethanoid glycosides as the alternative antioxidants and antimicrobial agents of natural origin.

The aim of the study was to develop an alternative capillary zone electrophoresis (CZE) for simultaneous determination of phillyrin (1), phillygenin (2), epipinoresinol-4-O-β-glucoside (3), pinoresinol-4-O-β-glucoside (4), lariciresinol (5), pinoresinol (6), isolariciresinol (7) and vladinol D (8) in Forsythia suspensa. The structural types of lignans 1–8 could be attributed to bisepoxylignans (1–4 and 6), monoepoxylignans (5 and 8) and cyclolignan (7). The major difficulties in the CZE separation of 1–8 could be summarization as the simultaneous presence of free lignans (1, 2 and 5–8) and lignan glucosides (3 and 4) and simultaneous occurrence of two pairs of isomers (3 and 4 as well as 5 and 7). Without the addition of β-CD and methanol, the resolution of these analytes was quite poor. However, in this study, compounds 1–8 were excellently separated from each other within 15 min under optimized conditions with a borax running buffer (40 mM borax, pH 10.30) containing 2 mM β-CD and 5% methanol (v/v) at the voltage of 20 kV, temperature of 35 °C and detection wavelength of 234 nm. Validation of the method included tests of linearity, precision, repeatability, stability and accuracy. In addition, the method offered inherent advantages such as lower analytical cost, no need of specific columns and use of small amounts of organic solvents and reagents. Finally, this green and economic CZE was successfully applied for the determination of these bioactive components 1–8 in F. suspensa fruits and its commercial extracts.