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.

Study on effect of gypenosides on insulin sensitivity of rats with diabetes mellitus via regulating NF-κB signaling pathway / 中国中药杂志

This study focused on the ameliorative effects of gypenosides(GPS) on insulin sensitivity and inflammatory factors in rats with type 2 diabetes mellitus(T2 DM) and explored their possible molecular mechanisms. After the successful establishment of T2 DM model, diabetic rats were randomly divided into four groups, including model group, GPS groups(200, 100 mg·kg~(-1)) and metformin group(100 mg·kg~(-1)), with healthy rats serving as the control. After 6-week intragastric administration, fasting blood glucose(FBG) and oral glucose tolerance were examined. The levels of insulin, C-peptide, tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), interleukin-6(IL-6) and C-reactive protein(CRP) in serum were examined. Then the homeostasis model assessment of insulin resistance(HOMA-IR) and insulin sensitivity index(ISI) were calculated. The protein expression levels of phosphorylated insulin receptor substrate-1(p-IRS-1) and phosphorylated protein kinase B(p-Akt) in skeletal muscle were measured by Western blot, as well as those of phosphorylated inhibitor of nuclear factor-κB(NF-κB) kinase β(p-IKKβ), phosphorylated alpha inhibitor of NF-κB(p-IκBα) and phosphorylated p65 subunit of NF-κB(p-p65) in adipose tissue. The relative expression levels of glucose transporter 4(GLUT4) mRNA in skeletal muscle and NF-κB mRNA in adipose tissue were measured by qRT-PCR, and the morphological changes of pancreatic tissue were observed. Compared with the model group, the GPS groups witnessed significant decrease in FBG, marked amelioration of impaired oral glucose tolerance and significant increase in ISI. Further, the high-dose GPS group saw significantly reduced HOMA-IR, TNF-α, IL-1β and CRP, significantly increased expression levels of p-IRS-1(Tyr), p-Akt and GLUT4, and markedly inhibited p-IRS-1(Ser), p-IKKβ, p-IκBα, p-p65 and NF-κB. The concentration of CRP and the expression levels of p-IRS-1(Ser), p-IKKβ, p-IκBα and NF-κB were remarkably reduced in the low-dose GPS group. However, GPS was found less effective in the regulation of serum insulin, C-peptide and IL-6 levels and the alleviation of pancreatic islet injury. The results indicated that GPS can reduce FBG and improve insulin sensitivity in diabetic rats possibly by regulating the NF-κB signaling pathway, inhibiting inflammation, and thereby regulating the expression of key proteins in the insulin signaling pathway.

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.

Qualitative analysis of Gynostemma longipes for medicinal usage / 中国中药杂志

The Qinling-Daba Mountains area is the main producing areas of Gynostemma longipes for medicinal usage, and samples of wild whole plants in Pingli, Shaanxi Province and Qingchuan, Sichuan Province were collected. The ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UHPLC-Q-TOF-MS~E) was used to profile the chemical compositions and analyze the similarities and differences of G. longipes samples in these areas. Based on the accurate molecular weight and fragment information obtained from Q-TOF-MS~E, the structures of the main components were identified by combining with the mass spectra, chromatographic behaviors of reference standards and related literatures. The results showed that the components of wild G. longipes from different places among Qinling-Daba Mountains area were similar. Forty-five chemical components were identified in the whole plant of G. longipes from Pingli, Shaanxi Province, including 43 triterpenoid saponins and 2 flavonoids which contain all main peaks in its fingerprint. The main components are dammarane-type triterpenoid saponins, such asgypenoside ⅩLⅨ, gypenoside A and its malonylated product of glycosyl.

Study on quality standard of gypenosides extract and Gypenosides Tablets / 中国中药杂志

In response to no national standard for Gynostemma pentaphyllum, a market survey was carried out, and 17 batches of gypenosides extract and 29 batches of Gypenosides Tablets on the market were collected. With gypenoside A as an index, the TLC qualitative identification and HPLC quantitative evaluation method of gypenosides extract and tablets was established. Based on the determination results of 17 batches of gypenosides extract and 29 batches of Gypenosides Tablets, the quality standards of gypenosides extract and tablets were formulated respectively, so as to give suggestions for improving the quality standards of gypenosides extract and tablets. Compared with the existing ministerial standards, the qualitative identification and quantitative detection of specific components were added, in order to provide scientific basis and suggestions for the revision of the quality standard of gypenosides extract and tablet preparation.

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.

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.

Analysis phylogenetic relationship of Gynostemma (Cucurbitaceae) / 中国中药杂志

The sequences of ITS, matK, rbcL and psbA-trnH of 9 Gynostemma species or variety including 38 samples were compared and analyzed by molecular phylogeny method. Hemsleya macrosperma was designated as outgroup. The MP and NJ phylogenetic tree of Gynostemma was built based on ITS sequence, the results of PAUP phylogenetic analysis showed the following results: (1) The eight individuals of G. pentaphyllum var. pentaphyllum were not supported as monophyletic in the strict consensus trees and NJ trees. (2) It is suspected whether G. longipes and G. laxum should be classified as the independent species. (3)The classification of subgenus units of Gynostemma plants is supported.

Preparation of novel gypenosides long-circulating liposomes consisted by sphingomyelin and beta-sitosterol modified by PEG / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the feasibility of preparing novel gypenosides long-circulating liposomes with PEG grafted on beta-sitosterol (PEG-Sito).</p><p><b>METHOD</b>Succinicanhydride was adopted to connect beta-sitosterol and PEG 2000. Sphingomyelin and PEG-Sito was used as material to prepare gypenosides long-circulating liposomes by using ethanol injection method. Encapsulation efficiency was determined by using protamine precipitation method. H-NMR was used to verify the synthesis of PEG-Sito, the novel gypenosides long-circulating liposomes were characterized by particle size, zeta potential and atomic force microscope.</p><p><b>RESULT</b>The synthesis of PEG-Sito was verified by 1H-NMR. Encapsulation efficiency of long-circulating liposomes prepared by ethanol injection method was 74.3%, particle size was 288.1 nm, zeta potential was -20.25 mV, the morphology were round observed by AFM.</p><p><b>CONCLUSION</b>The novel gypenosides long-circulating liposomes prepared with PEG-Sito was feasible, it had a high encapsulation efficiency and good morphology.</p>

Study on preparation of gypenosides liposomes composed of sphingomyelin and cholesterol and its quality evaluation / 中国中药杂志

<p><b>OBJECTIVE</b>To discuss the feasibility of preparing gypenosides liposomes with sphingomyelin and cholesterol, and optimize the preparation process and prescription.</p><p><b>METHOD</b>Gypenosides liposomes were prepared with sphingomyelin and cholesterol. The entrapment efficiency was determined by the protamine precipitation method. The entrapment efficiency was taken as the evaluation index to screen out the optimum preparation process for the new-type gypenosides liposomes. The preparation process was optimized by the orthogonal design. The new-type gypenosides liposomes were characterized by grain size, potential and atomic force microscope (AFM).</p><p><b>RESULT</b>Ethanol injection was the optimum process to prepare gypenosides liposome with sphingomyelin and cholesterol as follow: the ratio of gypenosides to sphingomyelin was 1: 10, the ratio of sphingomyelin to cholesterol was 4: 1, the pH of PBS buffer solution was 7.0, the hydration temperature was 45 degrees C, the entrapment efficiency was 79.06%, particle size was 191.4 nm, the Zeta potential was -33.16 mV, and the morphology were round under AFM.</p><p><b>CONCLUSION</b>It was feasible to prepare gypenosides liposome with sphingomyelin and cholesterol. The gypenosides liposomes prepared by the optimum preparation process were good in morphology, particle size and reproducibility.</p>

Effect of gypenosides on DMN-induced liver fibrosis in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of gypenosides on DMN-induced liver fibrosis in rats.</p><p><b>METHOD</b>A rat liver fibrosis model was established by injecting DMN intraperitoneally. Four weeks later, model rats were randomly devided into three groups: the model group, the gypenosides treated group (200 mg x kg(-1)) and the colchicine treated group (0.1 mg x kg(-1)), with 10 specimens for each group. After a 2-week treatment, following parameters were observed: (1) last body weight, weight ratio between liver and spleen; (2) content of liver hydroxyproline (Hyp); (3) activity of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyltransferase (gamma-GT), content of albumin (Alb) and total bilirubin( TBiL) in serum; (4) liver pathology (Sirius red staining and HE staining); (5) activity of liver superoxide dismutase (SOD), glutathione reduced (GSH), glutathione peroxidase (GSH-Px) and content of liver maleic dialdehyde (MDA).</p><p><b>RESULT</b>There were classic liver cirrhosis pathological changes in model groups. Compared with the normal group, liver Hyp content, activity of serum ATL, AST, gamma-GT and content of serum TBiL, MDA of model groups significantly increased; content of serum Alb and liver GSH, activity of liver SOD and GSH-Px decreased significantly in model groups. In comparison with the model group, liver cirrhosis remarkable improved in the gypenosides group, content of liver Hyp reduced significantly (P < 0.01), which was equal to the colchicine group. Compared with the model group, liver function parameters improved markedly in the gypenosides group; liver SOD and GSH-Px activities significantly increased; MDA content reduced significantly (P < 0.05).</p><p><b>CONCLUSION</b>Gypenosides shows an effect in treating DMN-induced liver fibrosis in rats.</p>

Uniform designed research on the active ingredients assembling of Chinese medicine prescription for anti-liver fibrosis / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the method and significance for studying active anti-liver fibrosis ingredients consisted Chinese medicine compound prescription based on Chinese medicine theory.</p><p><b>METHODS</b>Optimized prescription was screened out, adopting uniform block design with 4-factor 8-level table and regression analysis, through applying the four known effective ingredients (cordyceps sinensis polysaccharide, salvianolic acid B, amygdaloside and gypenosides) of Fuzheng Huayu Capsule (FZHYC, a new Chinese medine anti-liver fibrosis drug) to two rat liver fibrosis models established separately by dimethylnitrosamine (DMN) and CCl4, and taking the liver content of hydroxyproline (Hyp) as the screen index. Then a further study for comparing and verifying the efficacy of the obtained optimized prescription was conducted on the two former models respectively by observing the changes of Hyp content in liver, serum ALT activity and fibrosis pathology after medication, controlled by the original FZHYC and the recipe assembled by all the four ingredients.</p><p><b>RESULTS</b>Two optimized prescriptions (OPA and OPB) were screened out separately in the studies conducted on the two models. Both of them were consisted of cordyceps sinensis polysaccharide, Amygdaloside and Gypenosides, but different in constituent ratio, i.e., the ratio in OPA was 60 : 80: 50, and that in OPB, 20: 160: 50. Verifying study showed both OPA and OPA were significantly effective, with the efficacy equivalent to that of FZHYC (P>0.05). However, when they were used in combining with salvianolic acid B (the cutout ingredient in the screening), the efficacy lowered surely.</p><p><b>CONCLUSIONS</b>Uniform design is a valuable method in the compatibility research of Chinese Medicine drugs' composition. To assemble a new compound recipe reasonably based on the prescription of traditional compound recipe could make its effect equivalent to that of the original prescription. Ingredients or constituents in a prescription, either presented synergistic or antagonistic effects, are not randomly stacked together, and they should be orderly assembled in intrinsic rules of qualitative and quantitative changing.</p>

Effect and mechanism of gypenoside on the inflammatory molecular expression in high-fat induced atherosclerosis rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To evaluate the effect and possible mechanism of gypenoside (GP) on expression of inflammatory factors in aortic lesion of rats with high-fat induced atherosclerosis.</p><p><b>METHODS</b>Atherosclerotic rat model was established by feeding high-fat diet and intraperitoneal injection of vitamin D3. Sixty healthy male SD rats were randomly divided into the normal group, the model group, the simvastatin treated group and the three GP groups treated respectively with different dosages of GP. Rats were sacrificed 7 weeks later, their histopathological changes in thoracic aorta were observed by light microscope; expressions of intercellular adhesion molecule 1 (ICAM-1), monocyte chemotactic protein-1 (MCP-1) and nuclear factor-kappaBp65 (NF-kappaBp65) in aortic wall were detected by immunohistochemistry; serum level of oxidized low-density lipoprotein (ox-LDL) was determined by ELISA; serum total antioxidant capacity determined by colorimetry, and serum malondialdehyde (MDA) level determined by Thiobarbituric acid method.</p><p><b>RESULTS</b>In comparing with the model group, GPS showed actions in lessening the atherosclerosis lesion; reducing expressions of ICAM-1, MCP-1 and NF-kappaBp65 in aortic wall (P<0.01) and serum levels of MDA, ox-LDL (P < 0.01), as well as increasing the serum level of total antioxidant capacity (P < 0.01 ).</p><p><b>CONCLUSION</b>GP can down-regulate the expressions of ICAM-1 and MCP-1, inhibit the atherosclerosis formation in experimental rats, its mechanism might be related with its anti-oxidation effect and further inhibiting on the NF-kappaB activation.</p>

Identification of Gynostemma from their adulterant Cayratia japonica by PCR-RFLP / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a convenient and effective method for the identification of Gynostemma and Cayratia japonica.</p><p><b>METHOD</b>Eight species, including Gynostemm pentaphyllum, G. pentagynum, G. cardiospermum, G. longipe, G. yixingense, G. laxiflorum, G. guangxiense and C. japonica were investigated through PCR - RFLP of six chloroplast DNA fragments. The six gene fragments were digested by six restriction endonuclease respectively, including Taq I, Hpa II, EcoR I, Rsa I, Hha I, Hind III.</p><p><b>RESULT</b>Seven species of Gynostemma and their adulterant could be identified by trnK1f-trnK2r and Rsa.</p><p><b>CONCLUSION</b>PCR - RFLP provides a quick, reliable molecular marker technique for identification of Cynostemma and their adulterant Cayratia japonica.</p>

The effect of Gynostemma pentaphyllum (GP) on plasma lipoprotein metabolism and lipoperoxidation lipoprotein in the experimental hyperglycemia rats / 中国应用生理学杂志

<p><b>AIM</b>To study the regulation mechanism of GP on plasma lipoprotein metabolism and To explore its mechanism of anti-lipoperoxidation in the experimental hyperglycemia rats.</p><p><b>METHODS</b>The rats were raised with high fat diet for six weeks,and the model of hyperglycemia was then established. After that, those rats were treated with high or low dose of GP and xuezhikang as a masculine comparison for four weeks. Then, those rat were executed, and detected the plasma TC, TG, LDL-C, HDL-C, GSH-Px, at the same time the SOD, CAT and MDA concentration also be mensurated.</p><p><b>RESULTS</b>The results showed that high and low dose of GP could decrease the concentration of serum LDL-C, cholesterol and triglyceride remarkably and raise the concentration of HDL-C. The activity of GSH-Px, SOD and CAT in GP groups were promoted and the level of MDA was decreased distinctly.</p><p><b>CONCLUSION</b>The GP can therapy the abnormity of serum lipid and has obviously anti-lipoperoxidation affection.</p>

Analysis of the inhibitory effect of gypenoside on Na(+), K (+)-ATPase in rats' heart and brain and its kinetics / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the effects of gypenoside (Gyp) on the activity of microsomal Na(+), K(+)-ATPase in rat's heart and brain in vitro.</p><p><b>METHODS</b>The microsomal Na(+), K(+)-ATPase was prepared from rat's heart and brain by differential centrifugation. The activity of microsomal Na(+), K(+)-ATPase was assayed by colorimetric technique. Enzyme kinetic analysis method was used to analyze the effect of Gyp on the microsomal Na(+), K(+)-ATPase of rats.</p><p><b>RESULTS</b>Gyp reversibly inhibited the brain and heart's microsomal Na(+), K(+)-ATPase in a concentration-dependent manner, and showed a more potent effect on enzyme in the brain. The IC(50) of Gyp for the heart and brain were 58.79+/-8.05 mg/L and 52.07+/-6.25 mg/L, respectively. The inhibition was enhanced by lowering the Na(+), or K(+) concentrations or increasing the ATP concentration. Enzyme kinetic studies indicated that the inhibitory effect of Gyp on the enzyme is like that of competitive antagonist of Na(+), the counter-competitive inhibitor for the substrate ATP, and the mixed-type inhibitor for K(+).</p><p><b>CONCLUSION</b>Gyp displays its cardiotonic and central inhibitory effects by way of inhibiting heart and brain's microsomal Na(+), K(+)-ATPase activities in rats.</p>

Effect of total flaveos of Gymostemma pentaphyllum on protein expression of Fas/FasL genes and TNF-alpha concentration in cultured neonatal rat cardiomyocytes with hypoxia-reoxygenation / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of total flaveos of Gymostemma pentaphyllum on the protein expression of apoptosis-associated Fas/FasL gene and tumor necrosis factor-alpha (TNF-alpha) concentration in cultured neonatal rat cardiomyocytes with hypoxia-reoxygenation (H/R).</p><p><b>METHOD</b>A cultured primary neonatal rat cardiomyocytes model with H/R was erected, experiments were divided into six groups, (1)control group, (2)H/R group, (3)15 mg x L(-1) TFG plus H/R group, (4)45 mg x L(-1) TFG plus H/R group, (5) 105 mg x L(-1) TFG plus H/R group, (6)105 mg x L(-1) TFG group. TNF-aconcentration in cultured cardiomyocytes with H/R, was determined by ELISA method, the protein expression of Fas/FasL genes were estimated by immunohisto-chemistry.</p><p><b>RESULT</b>After cardiomyocytes were made with H/R, Compared with control group, the positive expression index (PEI) of Fas/FasL proteins in cardiomyocytes increased significantly, Compared with H/R groups, the PEI of Fas/FasL proteins were lower significantly in groups with different dosages of TFG (P < 0.05). TFG inhibited the secretion of TNF-alpha from myocardial cells and increased the survival rate of myocardial cells.</p><p><b>CONCLUSION</b>The protein expression of apoptosis-associated Fas/FasL genes increased during H/R. The TFG can protect myocardium against H/R injury by decreasing the production of TNF-alpha, downregulating the protein expression of Fas/FasL genes, and then inhibiting myocyte apoptosis.</p>