Identification of terpene synthase gene family in Gynostemma pentaphyllum and expression pattern analysis under abiotic stresses / 中国中药杂志

The present study aimed to investigate the composition of the terpene synthase(TPS) gene family in Gynostemma pentaphyllum and its role in abiotic stresses. The G. pentaphyllum TPS gene family was identified and analyzed at the genome-wide level using bioinformatics analysis, and the expression patterns of these family members were analyzed in different tissues of G. pentaphyllum as well as under various abiotic stresses. The results showed that there were 24 TPS gene family members in G. pentaphyllum with protein lengths ranging from 294 to 842 aa. All of them were localized in the cytoplasm or chloroplasts and unevenly distributed on the 11 chromosomes of G. pentaphyllum. The results of the phylogenetic tree showed that the G. pentaphyllum TPS gene family members could be divided into five subfamilies. As revealed by the analysis of promoter cis-acting elements, TPS gene family members in G. pentaphyllum were predicted to respond to a variety of abiotic stresses such as salt, low temperature, and dark stress. The analysis of gene expression patterns in different tissues of G. pentaphyllum revealed that nine TPS genes were tissue-specific in expression. The qPCR results showed that GpTPS16, GpTPS17, and GpTPS21 responded to a variety of abiotic stresses. This study is expected to provide references in guiding the further exploration of the biological functions of G. pentaphyllum TPS genes under abiotic stresses.

Gynostemma pentaphyllum saponins alleviate non-alcoholic fatty liver disease in rats by regulating intestinal flora and short-chain fatty acid metabolism / 中国中药杂志

This study aimed to explore the effects of Gynostemma pentaphyllum saponins(GPs) on non-alcoholic fatty liver disease(NAFLD) induced by high-fat diet in rats and reveal the underlying mechanism. The NAFLD model rats were prepared with high-fat diet. Forty male Sprague Dawley(SD) rats were randomly assigned into the control group, model group, and low-, moderate-, and high-dose GPs(50, 100, and 150 mg·kg~(-1), respectively) groups. After intragastric administration for 8 continuous weeks, we determined the body weight, liver weight, the levels of total cholesterol(TC), triglyceride(TG), low-density lipoprotein cholesterol(LDL-c), high-density lipoprotein cholesterol(HDL-c), alanine aminotransferase(ALT), and aspartate aminotransferase(AST) in serum, and the levels of TC, TG, malondialdehyde(MDA), superoxide dismutase(SOD), catalase(CAT), and interleukin 6(IL-6) in the liver. Furthermore, we observed the pathological changes of liver tissue by oil red O staining and hematoxylin-eosin(HE) staining, sequenced the 16 S rRNA of the intestinal flora in rat feces, and determined the content of short-chain fatty acids in rat feces. The results showed that GPs inhibited the excessive weight gain of high-fat diet-induced NAFLD in rats, reduced the liver weight, lowered the TC, TG, LDL-c, AST, and ALT levels in serum(P<0.05), and rose the HDL-c level in serum(P<0.01). GPs relieved the liver damage caused by high-fat diet, mainly manifested by the lowered levels of TC, TG, MDA, and IL-6 in the liver(P<0.01) and elevated levels of CAT and SOD in the liver. Furthermore, GPs reversed the intestinal flora disorder caused by high-fat diet, restored the diversity of intestinal flora, increased the relative abundance of Bacteroides, and reduced the relative abundance of Firmicutes and the ratio of Firmicutes to Bacteroides. Moreover, GPs promoted the proliferation of beneficial bacteria such as Akkermansia, Bacteroides, and Parabacteroides, and inhibited the growth of harmful bacteria such as Desulfovibrio, Escherichia-Shigella, and Helicobacter. GPs increased the content of short-chain fatty acids(acetic acid, propionic acid, and butyric acid)(P<0.01). These findings indicate that GPs can alleviate the high-fat diet-induced NAFLD in rats via regulating the intestinal flora and short-chain fatty acid metabolism.

Simultaneous quantitative analysis of nine saponins in Gynostemma pentaphyllum before and after heat processing based on UPLC-Q-Trap-MS / 中国中药杂志

Heat-processed Gynostemma pentaphyllum has strong biological activity, and saponins are the main components. To investigate the changes of saponins in G. pentaphyllum before and after heat processing, the present study determined and analyzed the content of nine saponins in G. pentaphyllum from Zhangzhou of Fujian and Jinxiu of Guangxi by ultra-high performance liquid chromatography with quadrupole ion-trap mass spectrometry(UPLC-Q-Trap-MS). The separation of the analytes was performed on an ACQUITY UPLC BEH C_(18) column(2.1 mm×50 mm, 1.7 μm) at 30 ℃, with acetonitrile and 0.1% formic acid in water as the mobile phase by gradient elution, and the flow rate was 0.3 mL·min~(-1). Quantitative analysis was performed using electrospray ionization source(ESI) in the multiple reaction-monitoring(MRM) mode. The results showed that the content of saponins with biological activities increased after heat processing. Specifically, gypenoside L, gypenoside LI, damulin A, damulin B, ginsenoside Rg_3(S), and ginsenoside Rg_3(R) in G. pentaphyllum produced in Zhangzhou of Fujian increased by 7.369, 8.289, 12.155, 7.587, 0.929, and 1.068 μg·g~(-1), respectively, while the content of ginsenoside Rd, gypenoside LVI, and gypenoside XLVI, which were abundant in the raw materials, decreased by 0.779, 19.37, and 9.19 μg·g~(-1), respectively. The content of gypenoside L, gypenoside LI, damulin A, damulin B, ginsenoside Rg_3(S), and ginsenoside Rg_3(R) in G. pentaphyllum produced in Jinxiu of Guangxi increased by 0.100, 0.161, 0.317, 0.228, 3.280, and 3.395 μg·g~(-1), respectively, while the content of ginsenoside Rd, gypenoside LVI, and gypenoside XLVI in the raw materials was reduced by 1.661, 0.014, and 0.010 μg·g~(-1), respectively. The results suggest that heat processing is an effective way to transform rare gypenosides. Furthermore, it is found that there are great differences in the content of gypenosides in different regions.

Mechanism of Gynostemma pentaphyllum in treatment of metabolism associated fatty liver disease based on network pharmacology and molecular docking / 中国中药杂志

The present study explored the mechanism of action of Gynostemma pentaphyllum in the treatment of metabolism associa-ted fatty liver disease(MAFLD) by network pharmacology and molecular docking. The main active components and action targets of G. pentaphyllum were collected from TCMSP. Disease-related targets were obtained from GeneCards, OMIM and TTD, and the common targets of the three databases were screened out, which were converted to the genes with standard names by UniProt. The drug-disease common target genes were obtained through Venn tool and uploaded to STRING for the construction of the protein-protein interaction(PPI) network. Cytoscape was used to construct and analyze the drug-active component-common target-disease network. The gene ontology(GO) analysis and Kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analysis were performed on the common targets by DAVID. Pymol was adopted to perform molecular docking of active components and the common targets and predict their binding ability. Twenty-four active components(such as gypenosides, quercetin and sitosterol) of G. pentaphyllum were screened out. Ninety-two targets were obtained and 54 common targets were identified. Key targets included TNF, IL6, PTGS2, TP53, CCL2 and VEGFA. GO analysis on biological processes, molecular functions and cellular components and KEGG pathway analysis were performed, and the results indicated that NF-κB, PI3 K-Akt, TNF and HIF-1 signaling pathways were mainly involved. Molecular docking results showed that gypenosides and quercetin had a strong binding ability to TNF, IL6 and PTGS2. The findings of this study revealed that the therapeutic efficacy of G. pentaphyllum on MAFLD might be achieved by resisting inflammation and oxidative stress and improving insulin resistance, providing ideas and a theoretical basis for the development and application of G. pentaphyllum in the treatment of MAFLD.

A novel dammarane-type saponin from Gynostemma pentaphyllum and its neuroprotective effect / 中国中药杂志

One new and two known dammarane-type saponins were isolated from the leaves of Gynostemma pentaphyllum using various chromatographic methods. Their structures were identified by HR-ESI-MS,~( 1)H-NMR, ~(13)C-NMR, 2 D-NMR spectra as 2α,3β,12β,20,24(S)-tetrahdroxydammar-25-en-3-O-[β-D-glucopyranosyl(1→2)-β-D-glucopyranosyl]-20-O-β-D-xylopyranosyl(1→6)-β-D-glucopyranoside(1, a new compound, namely gypenoside J5) and 2α,3β,12β,20,24(R)-tetrahdroxydammar-25-en-3-O-[β-D-glucopyranosyl(1→2)-β-D-glucopyranosyl]-20-O-β-D-xylopyranosyl(1→6)-β-D-glucopyranoside(2) and 2α,3β,12β,20-tetrahydroxy-25-hydroperoxy-dammar-23-en-3-O-[β-D-glucopyranosyl(1→2)][β-D-glucopyranosyl]-20-O-[β-D-xylopyranosyl(1→6)]-β-D-glucopy-ranoside(3), respectively. Compounds 1 and 2 were a pair of C-24 epimers. All compounds showed weak cytotoxicity agxinst H1299, HepG2, PC-3, SH-SY5 Y cancer cell lines. However, they exerted protective effect against SH-SY5 Y cellular damage induced by H_2O_2 dose-dependently, of which compound 1 displayed the strongest antioxidant effect. The present study suggested that G. pentaphyllum has antioxidative potential and the saponins from G. pentaphyllum are considered as the active compounds with neuroprotecitve effect.

Study on the mechanism of Gynostemma pentaphyllum intervention on atherosclerosis based on network pharmacology / 国际中医中药杂志

Objective:To explore the mechanism of Gynostemma pentaphyllum in the treatment of atherosclerosis (AS) based on network pharmacology. Methods:TCMSP was used to analyze the chemical components and targeted effect of Gynostemma pentaphyllum, through the database like OMIM, TTD, drugbank and digsee to predict and screen the targeted effect of AS. The genes corresponding to the target were queried by UniProt database, and then the compound target (gene) network and protein-protein interaction network (PPI) were constructed by using Cytoscape 3.2.1 to screen out the core target. Finally, the function enrichment analysis of Gene Ontology (GO) and the pathway enrichment analysis based on Kyoto Encyclopedia of genes and genomes (KEGG) were carried out by David, and the mechanism of action was studied. Results:The compound-target network consisted of 13 compounds and 150 corresponding targets. The key targets were PGR, NR3C2, NCOA2, PPARG, PTGS1, PTGS2, etc. PPI network contains 131 proteins and 46 core proteins. There are 480 GO item in GO function enrichment analysis, including 403 entries in biological process (BP), 35 entries in cell composition (CC), and 42 entries in molecular function (MF). 25 signaling pathways related to AS were obtained by enrichment and screening of KEGG pathway, involving PI3K-AKT, TNF, HIF-1, MAPK, toll like receptor and other signaling pathways.Conclusions:This paper discussed the mechanism of Gynostemma pentaphyllum in the treatment of AS through network pharmacology, which provides new ideas and methods for further research and exploration of the mechanism of Gynostemma pentaphyllum in the treatment of AS.

Study on anti-hyperglycemia effect of aqueous extract from Gynostemma pentaphyllum leaves on diabetes rats / 中草药

Objective: The aim of this study was to investigate the anti-hyperglycemia effect and mechanism of aqueous extract from Gynostemma pentaphyllum (GP) leaves on STZ-induced diabetic rats. Methods: Diabetic rat models were established by intraperitoneal injection of streptozotocin (STZ, 50 mg/kg). A total of 21 successful SD male rats were randomly divided into model group (STZ). G. pentaphyllum leaves aqueous extract low dose group (GP•H2O-L, 100 mg/kg) and high dose group (GP•H2O-H, 500 mg/kg), another seven normal rats were taken as the control group. Blood samples were taken from the 2nd and 3rd weekends to detect plasma glucose and triglyceride concentrations; Real-time PCR was used to detect the expression of TNF-α and GLUT-4 mRNA in skeletal muscle; Western blotting was used to detect GLUT-4 total protein in skeletal muscle; The expression of GLUT-4 protein on skeletal muscle sarcolemma was observed by immunofluorescence staining. Results: The results showed that the food intake and water intake of the STZ group were significantly increased compared with the control group, while the body weight and skeletal muscle weight were obviously decreased; Plasma triglyceride and blood glucose concentrations and the expression of TNF-α mRNA in skeletal muscle were significantly increased, while the expression of GLUT-4 mRNA, GLUT-4 total protein and GLUT-4 protein in skeletal muscle sarcolemma was obviously decreased. Compared with the STZ group, the high-dose aqueous extract of G. pentaphyllum leaves significantly reduced the blood glucose of STZ-induced diabetic rats, and reversed the expression of TNF-α mRNA and GLUT-4 protein in skeletal muscle. Conclusion: The aqueous extract from G. pentaphyllum leaves could reduce hyperglycemia in STZ-induced diabetic rats, and its mechanism may be related to increasing the expression of GLUT-4 protein on skeletal muscle sarcolemma and inhibiting skeletal muscle inflammation.

Triterpenoids from Gynostemma pentaphyllum and their inhibition activity to α-glycosidase and protein tyrosine phosphatase 1B / 中草药

Objective: To determine the total saponins from Gynostemma pentaphyllum, the dammarane-type triterpenoids of its hydrolysate, and its hypoglycemic activity. Methods: Compounds from the acid hydrolyzate extracts and total saponins were isolated by silica gel, recrystal and preparative liquid chromatography, and their structures were identified by the NMR spectral analysis. The sensitive screening modles of α-glucosidase and PTP1B inhibitors were established in vitro. The inhibitory kinetics of compounds were also investigated. Using the method of computer aided drug design of active site, PTP1B interact with the strongest active compound for docking simulation. Results: Seven compounds were isolated from the acid hydrolyzate of total saponins, which identified as gpsapogenin A (1), 20(S)-panaxadiol (2), gypensapogenin F (3), 20(R)-protopanaxadiol (4), (23S)-3β- hydroxydama-20,24-diene-21-carboxylic acid 21,23-lactone (5), gypsapogenin A (6), and (20S,24S)-3β,20,21β,23β,25- pentahydroxy-21,24-epoxydammarane (7). Five compounds were isolated from total saponins, including (20R,23R)- 3β,20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-6-O- acetyl-β-D-glucopyranoside (8), (20S,23S)-3β,20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-6-O-acetyl-β-D-glucopyranoside (9), (20R,23R)-19-oxo-3β,20-dihydroxydammar-24-en-21-oci acid 21,23-lactone3-O-[α-L-rhamnopyranosyl-(1→2)][β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (10), (20S)-3β,20,21- trihydroxydammar-23,25-diene 3-O-{[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-β-D-glucopyranosyl}-21-O-β- D-glucopyranoside (11), and (20S,23S)-3β,20-dihydroxydammar-24-en-21-oic acid and 21,23-lactone 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl-(1→3)]-β-D-glucopyranoside (12). Conclusion: Beside compound 4, the other compounds showed inhibitory activity against α-glucosidase and PTP1B. For the α-glucosidase and PTP1B inhibitions assay, compound 9 indicated the strongest inhibitory effect with IC50 2.10 and 1.07 μmol/L, respectively.

Hangover relieving and antioxidant effects of Gynostemma pentaphyllum (Thunb.) Makino and/or Hovenia dulcis Thunb. extracts

The present study attempts to study alcohol metabolizing and antioxidant properties of Gynostemma pentaphyllum(Thunb.) Makino distillate (GPD) and combination effects with Hovenia dulcis Thunb. extract (HDE) on these activities.The alcohol-metabolizing activity of GPD with/without HDE was determined by assessing alcohol dehydrogenase(ADH) and acetaldehyde dehydrogenase (ALDH) activities. To define the effect of GPD with/without HDE on alcoholmetabolism, antioxidant activities and total phenolic content of GPD with/without HD extract were evaluated using2-diphenyl-1-picrylhydrazyl free radical scavenging, ferrous chelating assays, and the Folin–Ciocalteu method.Cytotoxicity against human normal liver CHANG cells was also evaluated using the 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. GPD treatment alone or in combination with HDE significantly increased ADHand ALDH activities; combined treatment was most effective. Total phenolic and flavonoid contents were greater incombination than the level found in GPD alone. GPD revealed a synergistic antioxidant effect when combined withHDE. GPD and/or HDE had no antiproliferative activity against the normal liver cell line. These results suggest thatGPD-HDE combination is the possible natural resource for the management of alcohol-induced liver injury.

Analysis of Gynostemma pentaphyllum sapoins from different regions based on LC-MS and pattern recognition method / 中草药

Objective: The fingerprint of Gynostemma pentaphyllum was established with liquid chromatograph-mass spectrometer (LC-MS), and the main chromatographic peaks were preliminary identified, and combined with multi-variable analysis pattern recognition method to identify G. pentaphyllum from different origins. Methods: Inertsil ODS-SP (250 mm × 4.6 mm, 5 μm) C18 column was eluted with acetonitrile-0.1% formic acid as mobile phase gradient. The detection wavelength was 203 nm. The mass spectrometry equipped with electrospray ionization (ESI) was used as detector and operated under the negative ion mode. A total of 21 batches of G. pentaphyllum were analyzed by cluster analysis, principal component analysis, and corresponding analysis. Results: The fingerprints of G. pentaphyllum were established by LC-MS, and nine common peaks and 10 characteristic peaks were calibrated. According to the mass spectrometry information and literature comparison, 16 chromatographic peaks were qualitatively analyzed. Combined with multivariate analysis, G. pentaphyllum from different habitats was distinguished. It would establish the foundations for quality control and comprehensive evaluation of G. pentaphyllum. Conclusion: The LC-MS and pattern recognition analysis can be used to distinguish G. pentaphyllum from different regions and the targeting compounds of interest would be separated.

Gypenoside granules improved lipid metabolism in C57BL/6J mice with hyperlipidemia / 中国药科大学学报

@#To investigate the hypolipidemic effects of gypenosides granules and its combination with lipitor, a model of hyperlipidaemia C57BL/6J mice was established by high-fat diet feeding for 4 weeks. The mice were randomly divided into blank group, model group, lipitor group(10 mg/kg of lipitor), low dose group(90 mg/kg of gypenosides granules), medium dose group(120 mg/kg of gypenosides granules), high dose group(180 mg/kg of gypenosides granules)and the combination group(180 mg/kg of gypenosides granules and 10 mg/kg of lipitor). After 4 weeks of continuous administration, the contents of serum lipid indexes, serum ALT, AST and apolipoprotein B(ApoB)were measured. The liver tissues of mice were observed by H&E staining. The expression levels of key factors involved in hepatic cholesterol metabolism were observed by RT-PCR and Western blot methods, such as adenosine triphosphate combined box transporter A1(ABCA1), liver X receptor(LXRα), cholesterol 7 alpha hydroxylase(CYP7A1)and type BΙ scavenger receptor(SR-BΙ). The results revealed that gypenosides granules significantly decreased the mice body weight, total abdominal fat area and the level of serum total cholesterol(TC). The combination group showed a more significant reduction in TC level than the other administration groups. Moreover, gypenosides granules treatment remarkably increased the protein expression of ABCA1 and up-regulated the mRNA expression of ABCA1, CYP7A1 and SR-BI. The above results suggest that gypenosides granules can significantly reduce blood lipid contents, and the combination therapy with lipitor show better the lipid-lowering effect. Meanwhile, gypenosides granules can decrease the level of serum transaminase. Preliminary exploration suggests the lipid-lowering mechanism of gypenosides granules may be involved in cholesterol reversal to regulate the level of TC.

Protective effects of flavonoids from Gynostemma pentaphyllum on oxidative damage in LLC-PK1 cells / 中国中药杂志

Four flavonoids were isolated from Gynostemma pentaphyllum by chromatography methods and their structures were identified by MS and NMR spectra data as quercetin-3-O-( 2″,6″-di-α-L-rhamnosyl)-β-D-galactopyranoside( 1),quercetin-3-O-( 2″,6″-di-α-L-rhamnosyl)-β-D-glucopyranoside( 2),quercetin-3-O-( 2″-α-L-rhamnosyl)-β-D-galactopyranoside( 3),and quercetin-3-O-( 2″-α-L-rhamnosyl)-β-D-glucopyranoside( 4). Among them,compounds 1-3 were obtained from the Cucurbitaceae family for the first time.The four flavonoids showed potent antioxidant effects against the DPPH,·OH and ■radicals in vitro,especially for DPPH radical scavenging activity with the IC50 values of 71. 4,29. 5,48. 3 and 79. 2 μmol·L~(-1),respectively. Moreover,the four flavonoids displayed strong cytoprotection against AAPH-induced oxidative damage in LLC-PK1 cells by suppressing the increase of malondialdehyde( MDA) and the decrease of the superoxide dismutase( SOD) and glutathione( GSH). Since further research is needed to prove its efficacy in vivo and clinical trial,the study may provide four potential antioxidants from G. pentaphyllum.

Simultaneous determination of eleven amino acids in Gynostemma pentaphyllum by UFLC-QTRAP-MS/MS / 中成药

AIM To establish a UFLC-QTRAP-MS/MS method for the simultaneous content determination of phenylalanine,isoleucine,methionine,valine,proline,tyrosine,aspartic acid,histidine,arginine,lysine and glutamic acid in Gynostemma pentaphyllum (Thunb.) Makino.METHODS The analysis of aqueous extract of G.pentaphyllum was performed on a Waters XBridge Amide column (2.1 mm × 100 mm,3.5 μm),with the mobile phase comprising of water-acetonitrile (containing 0.2％ formic acid) flowing at 0.6 mL/min in a gradient elution manner.RESULTS Eleven amino acids showed good linear relationships within their own ranges (r ＞0.998 9),whose average recoveries were 90.74％-103.05％ with the RSDs of 0.61％-5.00％.CONCLU-SION This accurate,stable and reproducible method can be used for the quality control of G.pentaphyllum.

Simultaneous Determination of Eight Saponins in Gynostemma pentaphyllum(Thunb.) Makino by HPLC / 广州中医药大学学报

Objective To establish a high performance liquid chromatography(HPLC) method for the simultaneous determination of eight components in Gynostemma pentaphyllum(Thunb.) Makino (ginsenosides Rb1, Rb3, Rd, Re, F2, and gypenosides A, XLIX and XVII). Methods HPLC was performed on Agilent Eclipse XDB-C18(4.6 mm × 250 mm, 5 μm) chromatographic column with 0.3％ formic acid solution (A)- acetonitrile (B) as the mobile phase by gradient elution, flow rate was 1.5 mL·min-1, detection wavelength was 203 nm, and column temperature was set at 50 ℃. Results There was a good linear relationship between peak area and concentration of eight components (r ≥ 0.999) , the recovery was in the range of 97％ to 100％. Conclusion The established method is simple, rapid, accurate, reliable, and reproducible, and can be used for the quality control of Gynostemma pentaphyllum(Thunb.) Makino.

Effects of flavonoids from Gynostemma pentaphyllum on A549 cells damaged by hydrogen peroxide / 中国中药杂志

This study focuses on the therapeutical effect of flavonoids from Gynostemma pentaphyllum on human lung carcinoma A549 cells induced by H₂O₂ oxidative stress and its possible mechanisms. The oxidative damage model was established using different concentrations H₂O₂ to induce A549 cell for different hours, and then treated with the flavonoids for 10 hours. The effects of flavonoids from G. pentaphyllum on cell viability of A549 cell damaged by H₂O₂ were detected by MTT assay. The contents of ROS were detected by DCFH-DA fluorescent probe method via flow cytometer. The contents of MDA, SOD and GSH were detected by TBA，NBT and DTNB-linked colorimetry assay, respectively. Expressions levels of Nrf2, NQO1 and HO-1 in A549 cells were evaluated by Western blot. The results showed that the cell activity was decreasing with the rise of H₂O₂ concentration within the range of 200-700 μmol·L⁻¹. The cell viability was 60.4% after treated with 500 μmol·L⁻¹H₂O₂ for 10 h, so it was chosen to be as an oxidant stress model. Compared with normal group，the contents of SOD, GSH and HO-1 expressions were lower after damaged with H₂O₂. On the contrary, the contents of ROS and MDA expressions were increased. Compared with model group, the contents of SOD, GSH and the expressions of Nrf2, NQO1 and HO-1 were increased after treated with flavonoids from G. pentaphyllum. The above results demonstrate that flavonoids from G. pentaphyllum may attenuate the effect of H₂O₂-induced oxidative stress on A549 cell by resisting oxidation. The finding may provide a biological evidence for the application of the G. pentaphyllum to fight the oxidative stress related diseases.

Comparison of saponins from Gynostemma pentaphyllum leaves prepared by different processing methods / 中国中药杂志

To investigate the differences of chemical compositions in Gynostemma pentaphyllum leaves prepared by different processing methods. Ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used to compare the chemical compositions between shade-dried processing and drum-dried processing. Forty six gypenosides were identified by control comparison， liquid chromatography-mass spectrometry(LC-MSn) fragmentation information， and literature data. The mass spectral peak area statistics was combined with principal component analysis(PCA)， and the results showed that eight batches of Gynostemma pentaphyllum leaves samples were divided into two groups according to the two different processing methods; ten chemical compositions with significant differences were screened according to mass spectrum information combined with partial least-squares discriminant analysis(PLS-DA). The result showed that most parent nucleus of the gypenosides contained three to four glycosides in drum-dried samples， and one to two glycosides in the shade-dried samples. It was inferred from further MS analysis that desugarization of gypenosides was present to produce secondary glycosides with the effect of glucosidase in the shade-drying， thus resulting in difference in compositions. This study provided data support for harvesting， processing and quality control of Gynostemma pentaphyllum leaves.

Effect of light quality on total gypenosides accumulation and related key enzyme gene expression in Gynostemma pentaphyllum / 中草药·英文版

Objective: Light quality has effect on the accumulation of gypenosides in the medicinal plant Gynostemma pentaphyllum in the family Cucurbitaceae, while the squalene synthase (SS) and squalene epoxidase (SE) are the key enzymes for gypenoside biosynthesis. The objective of this study was to elucidate the relationship between light quality and biosynthesis key enzyme involving the regulation of gypenoside accumulation. Methods: The content of total gypenosides was measured by colorimetric method and the expression of SS and SE gene was determined by quantitative Real-time PCR in the seedlings of G. pentaphyllum which were grown with different light quality. Results: Light quality showed remarkable impacts on the accumulation of total gypenosides. The highest content of total gypenosides in the plant under red light condition was determined, followed by blue light and white light, while the lowest content was recorded under dark condition. qRT-PCR analysis proved that the expression levels of SS and SE genes were also affected by light quality. The high-level gene expressions of SS and SE were found in the plant under red light condition, followed by blue light, with the least content in darkness. The statistical analysis revealed that the total gypenosides were significantly different in different light treatment and the content of total gypenosides was positively related to the expression of SS and SE genes. Conclusions: Light quality regulates gypenoside accumulation via altering the expression of SS and SE in G. pentaphyllum.

Research progress on chemical constituents in hydrolyzed products of gypenosides and their pharmacological activities / 药物评价研究

The biologically active constituents in Gynostemma pentaphyllum are dammarane-type glycosides,called gypenosides.They are believed to be the highest contents of this herb,easy to obtain,and mainly active in anti-tumor,controlling the blood glucose,lipid-lowering,cardiovascular protection,etc.The saponins may change into sub-glucoside after hydrolysis,for the intemal acetal glucoside structure is vulnerable to acid,alkali,and enzyme degradation.Through searching the literatures in recent years,this paper summarized the chemical constituents in the hydrolyzed products of gypenosides,for providing references to discover novel and more active lead compounds.

Research Progress in Chemical Constituents and Pharmacological Activities of Gynostemma pentaphyllum / 中国药学杂志

This paper summarized the chemical constituents and pharmacological activities of Gynostemma pentaphyllum based on systematic literature research. G. pentaphyllum mainly contains dammarane-type saponins and a total of 165 gynostemma saponins were obtained from 1976 to 2011.In recent years, some new gypenosides were isolated and identified. Two hundred and one gypenosides were classified and summarized in this paper. In addition, G. pentaphyllum also contains a variety of other chemical components including flavonoids, polysaccharides, amino acids, and trace elements, which are worth further research. The pharmacological effects of G. pentaphyllum including antitumor, regulating blood lipid, hypoglycemic, liver protection, anti-senility and improving immunity effects were summarized. This review will provide some information for further research of G. pentaphyllum.

Preventive Effect of Gynostemma pentaphyllum Water Extract on Model Rats with Water Immersion Stress-induced Gastric Ulcer / 中国药房

OBJECTIVE:To investigate the preventive effect of Gynostemma pentaphyllum water extract on model rats with stress-induced gastric ulcer. METHODS:60 rats were randomly divided into normal group,model group,positive group (Sanjiu weitai granules,3.69 g/kg)and G. pentaphyllum water extract high-dose,medium-dose,low-dose groups(4.0,2.0,1.0 g/kg,cal-culated by crude drug),10 in each group. All rats were intragastrically administrated,once a day,for one week. After 2 h of ad-ministration,except for normal group,rats were induced to stress gastric ulcer model by water immersion method in other groups. After 3 h of modeling,ulcer index (UI),ulcer inhibition rate were detected;superoxide dismutase (SOD) and malondialdehyde (MDA)in serum,tumor necrosis factor-α(TNF-α),prostaglandin E2(PGE2),nitric oxide(NO)and endothelin-1(ET-1)levels in gastric mucosal tissue were determined. RESULTS:Compared with normal group,SOD level in model group was decreased, while MDA level was increased in serum(P<0.05);PGE2,NO levels were decreased,TNF-α,ET-1 levels were increased in gas-tric mucosal tissue (P<0.05). Compared with model group,UI in administration groups were obviously decreased,above-men-tioned indexes were obviously improved,which showed certain dose-dependence. And the levels of PEG2,ET-1 of gastric mucosal tissue in G. pentaphyllum water extract high-dose group improved more obviously than positive group,with statistical significances (P<0.05). CONCLUSIONS:G. pentaphyllum water extract plays a role in preventing gastric ulcer by regulating serum SOD-MDA balance,gastric mucosal tissue NO-ET-1 balance and reduce the degree of gastric mucosa injury,etc.