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.

Therapeutic effects of gypenosides on hypercholesterolemia and it protective effect on liver injury / 中国药科大学学报

@#In order to explore the therapeutic effects and preliminary mechanism of gypenosides (GP) on hypercholesterolemia, as well as the protective effect on liver injury induced by high-dose simvastatin and high cholesterol diet (HCD), the hypercholesterolemia model of golden hamster was established by high cholesterol diet. The experimental animals were divided into blank group, model group, GP low and high dose groups (60 mg/kg, 120 mg/kg), simvastatin group (10 mg/kg), and GP high dose combined with simvastatin group (120 mg/kg + 10 mg/kg).The efficacy was investigated through dynamic monitoring serum cholesterol and liver function related indexes after drug treatment of 14 and 23 days. The results showed that GP could significantly reduce the levels of serum low density lipoprotein cholesterol (LDL-C), total cholesterol (TC), triglyceride (TG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP), increase the level of serum high density lipoprotein cholesterol (HDL-C), and reduce the secretion of PCSK9. It is suggested that GP has a good therapeutic effect on HCD diet-induced hypercholesterolemia hamsters, which may be related to its inhibition of PCSK9 secretion. In addition, GP can significantly ameliorate liver damage caused by HCD diet and high-dose simvastatin. These findings provide a scientific basis and useful reference for the combination of GP and statins to reduce toxicity and increase efficacy.

Mechanism by which gypenosides alleviate oxidative stress injury induced by h2o2 in rat osteoblasts / 中国组织工程研究

BACKGROUND: Gypenosides have antioxidant properties, with beneficial effects such as reducing blood pressure, anti-aging and anti-tumor, but the specific protective mechanism is not clear. It is also unknown whether gypenosides have effect on the proliferation and differentiation of oxidative stress-damaged osteoblasts. OBJECTIVE: To investigate the mechanism by which gypenosides alleviate oxidative stress injury in rat osteoblasts and the effect on the proliferation and differentiation of oxidatively damaged osteoblasts. METHODS: Monolayer cell culture method was used to separate neonatal rat skull cells for the culture of osteoblasts. In this experiment, there were three groups, with normal culture medium as blank group, normal culture medium+oxidative damage as control group, and normal culture medium containing gypenosides and oxidative damage as experimental group. Osteoblasts in the experimental and control groups were cultured in the culture medium containing 150 μmol/L H2O2. After 3 and 5 days of intervention, cell counting kit-8 method was used to detect the effects of gypenosides on oxidative damage of osteoblasts. Alkaline phosphatase staining was used to detect alkaline phosphatase activity on day 7 after induction. Alizarin red staining was used on day 21 of induction to observe osteoblast mineralization. Western blot was used to detect the expression of NOX4, bone morphogenetic protein 2 and Smad4. The experimental protocol was approved by the Animal Ethics Committee of Guangzhou University of Chinese Medicine. RESULTS AND CONCLUSION: Gypenosides could promote the proliferation of oxidatively damaged osteoblasts. The results of alkaline phosphatase staining and alizarin red staining showed that gypenosides could promote the differentiation of oxidatively damaged osteoblasts. Compared with the control group, gypenosides could downregulate the expression of NOX4 protein and upregulate the expression of bone morphogenetic protein 2 and Smad4 protein in the experimental group, with statistically significant results (P < 0.05). All these findings indicate that gypenosides have a protective effect on H2O2-induced oxidative stress injury in osteoblasts, and promote the proliferation and differentiation of damaged osteoblasts. The mechanism may be related to the inhibition of Nox4 protein expression and the activation of bone morphogenetic protein/Smad pathway.

Study on the Effects of Gypenosides on Gene Expression of Major Urinary Proteins in Hypercholesterolemia Model Mice / 中国药房

OBJECTIVE：To investigate the effects of gypenosides （GPs）on gene expression of major urinary proteins （Mups） in liver tissue of hypercholesterolemia model mice. METHODS ：C57BL/6J mice were divided into control （ND）group，model （HFD）group and GPs therapy （GP）group according to body weight （BW），with 11 mice in each group. Except for ND group ， other groups were given high-lipid diet to induce hypercholesterolemia model. From the 17th week of feeding ，ND group and HFD group were given constant volume of 0.1％CMC-Na solution intragastrically ；GP group were given GPs suspension （250 mg/kg） intragastrically，once a day ，for consecutive 22 weeks. BW ，the levels of blood glucose （BG）and blood lipid （TC，LDL-C）were detected in each group. Total RNA of liver tissue was extracted ，and reverse transcription library was constructed and RNA-seq sequencing was performed. The differentially expressed genes were screened by PCA ，volcano map and scatter plot. RT-qPCR was used for verification for differentially expressed genes. The correlation between the expression of differentially expressed genes and the above pharmacodynamic indexes was analyzed by bivariate analysis. RESULTS ：Compared with ND group ，BW，the levels of BG，TC and LDL-C in HFD group were increased significantly （P＜0.05）. Compared with HFD group ，above indexes of GP group were decreased significantly except for BW （P＜0.05）. PCA showed that the data of ND group and HFD group distributed in different quadrants ，and the data distribution of GP group was between above two groups. mRNA of Mup4，Mup5，Mup11，Mup15 and Mup21 in liver tissue of mice were increased significantly after treated with high-fat diet （P＜0.05）. mRNA of Mup3，Mup4， Mup5，Mup8，Mup12 and Mup21 were decreased significantly after treated with GPs （P＜0.05）. In ND group vs. HFD group and HFD group vs. GP group ，mRNA of Mup4，Mup5 and Mup21 genes changed significantly and the trend was opposite. Results of RT-qPCR verification showed that compared with ND group ，relative mRNA expression of Mup4，Mup5 and Mup21 gene were increased significantly in HFD group （P＜0.05）. Correlation analysis revealed that mRNA expression of Mup5 was positively correlated with the levels of TC and BG （r＝0.727 1，0.670 6，P＜0.05），mRNA expression of Mup4 and Mup21 were positively correlated with the level of BG （r＝0.737 8，0.721 5，P＜0.05）. CONCLUSIONS ：GPs can regulate the expression of Mups genes in liver tissue of hypercholesterolemia model mice ， and reduce glucose and lipid level through regulating the mRNA over-expression of Mup4，Mup5 and Mup21.

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.

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.

Gypenosides protect acute alcoholic liver injury through Nrf2/nf-kb signaling pathway in mice / 中国药理学通报

Aim To investigate the protective effect of gypenosides on acute alcoholic liver injury in mice and its mechanism. Methods The BALB/c mice were divided into five groups, control, alcoholic injury group and three protection groups by different doses of gypenosides. The mice in protection groups were given different doses of gypenosides everyday for one week. After the mice were sacrificed, the peripheral blood and the liver were collected. The pathology of liver tissue sections was observed by HE staining. The ALT, AST, TC and TG were determined by automatic biochemical analyzer. TNF-a and IL-6 mRNA levels were determined by real-time PGR. The content of MDA and GSH, the activities of SOD and CAT and the level of GSH were determined by kits respectively. The protein levels of proteins of Nrf2 and NF-kB signaling pathways were assayed by Western blot. Results Alcohol treatment increased the levels of ALT, AST, TC and TG as well as the expression levels of TNF-a and IL-6. At the same time, alcohol treatment increased the content of MDA and decreased the activities of SOD, CAT and GSH. The alcohol treatment also inhibited the Nrf2 signaling pathway and induced the NF-kB translocation to nuclear. The gypenoside supplementation significantly inhibited all the above changes in a dose-dependent way. Conclusion The gypenoside supplementation shows significant protection on the acute alcoholic liver oxidative injury in mice by regulating Nrf2/NF- kB signaling pathway.

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.

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.

Effects of gypenosides on PCSK9 gene expression and blood lipids lowered by simvastatin / 中国病理生理杂志

AIM:To explore the effect of gypenosides ( GPs) on PCSK9 gene expression in hyperlipidemic rat liver and the blood lipids lowered by simvastatin .METHODS: Healthy male SD rats ( n=60 ) were randomized into 5 groups:normal control group , hyperlipidemic model group , simvastatin group , GPs group and GPs combined with simvasta-tin group ( combined group ) .The rats in all groups were fed high-fat diet except normal control group which were fed with ordinary diet.The rats in control group and hyperlipidemic model group were gavaged with 0.3%CMC-Na every day.The rats in GPs group were gavaged with GPs at 160 mg? kg-1? d-1 .The rats in simvastatin group were gavaged with simvas-tatin at 5 mg? kg-1? d-1 .The rats in combined group were gavaged with GPs and simvastatin .The experiment lasted for 8 weeks.The rats were anesthetized with chloral hydrate , and abdominal arterial blood samples were collected to detect the total cholesterol ( TC) , triglyceride ( TG) , low-density lipoprotein cholesterol ( LDL-C) and high-density lipoprotein cho-lesterol ( HDL-C) .The body weight and the wet weight of the livers were measured , and the liver index was calculated . The pathological changes of the livers were observed under microscope with HE staining .The expression of PCSK9 and low-density lipoprotein receptor ( LDLR) at mRNA and protein levels was determined by real-time PCR and Western blot .RE-SULTS:The model of hyperlipidemia rats was established successfully .Compared with model group , the levels of TC , TG and LDL-C in simvastatin group, GPs group and combined group were obviously decreased (P<0.05), and the HDL-C levels were obviously upregulated (P<0.05).Compared with model group, the liver indexes in simvastatin group, GPs group and combined group were obviously decreased (P<0.05).The pathological changes of the liver tissues showed that hepatic adipose appeared in model group , and that in simvastatin group and GPs group had different degrees of relief , espe-cially in combined group .Compared with model group , the mRNA expression levels of PCSK 9 and LDLR in simvastatin group were obviously increased , while the mRNA expression levels of PCSK 9 in GPs group and combined group were obvi-ously decreased (P<0.05), and the mRNA expression of LDLR in combined group was obviously increased (P<0.05). Compared with model group , the protein expression of PCSK 9 and LDLR in simvastatin group was obviously increased , while the protein expression levels of PCSK 9 in GPs group and combined group were obviously reduced , and the LDLR pro-tein levels were obviously increased (P<0.05).CONCLUSION:Gypenosides inhibit the expression of PCSK9 and in-crease the expression of LDLR in the liver .The combination of gypenosides and simvastatin promotes the lipid-lowering effect of simvastatin and attenuates hepatic steatosis , which may be related to inhibiting the expression of PCSK 9 in the liv-er.

Effect of gypenosides on RAGE and TGF-β1 expression in human mesangial cells induced by AGEs / 中国药理学通报

Aim To observe the effect of gypenosides ( GP) on the expression of receptor for advanced gly-cated endproducts ( RAGE ) and transforming growth factor-β1 ( TGF-β1 ) in human mesangial cells( HMCs) induced by AGEs. Methods HMCs were cultured in DMEM of low glucose containing 15% fetal bovine ser-um in vitro, which were divided into four groups: the normal group, model group, GP group and positive control group. In addition to the normal group, the other groups were stimulated by AGEs ( 200 mg · L-1 );furthermore, GP group was intervened with dif-ferent concentrations(25,75,175 mg·L-1) of GP, while control group was given 10 mmol · L-1 of amin-oguanidine hydrochloride. The expression of RAGE and TGF-β1 protein of each group was detected by Western blot; the expression of RAGE and TGF-β1 mRNA of each group was detected by RT-PCR. Re-sults The expression of RAGE, TGF-β1 protein and mRNA in HMCs induced by AGEs in the model group was significantly higher than that of the normal group ( P<0. 01 );compared with the positive control group ( P<0. 01 ) , GP could obviously reduce the expression of RAGE, TGF-β1 protein and mRNA in a dose-de-pendent manner. Conclusion GP can reduce the ex-pression of RAGE in HMCs induced by AGEs, block AGEs-RAGE signaling pathway and decrease the ex-pression of the downstream factor TGF-β1 , therefore, it plays the role in the resistance of rennal fibrosis in DN.

Effects of Gypenosides on Dopaminergic Neuronal Cell Death in 6-Hydroxydopamine-lesioned Rat Model of Parkinson's Disease with Long-term L-DOPA Treatment

The goal of this study was to determine whether gypenosides (GPS) exert protective effects against dopaminergic neuronal cell death in a 6-hydroxydopamine (OHDA)-lesioned rat model of Parkinson's disease (PD) with or without long-term 3,4-dihydroxyphenylalanine (L-DOPA) treatment. Rats were injected with 6-OHDA in the substantia nigra to induce PD-like symptoms; 14 days after injection, groups of 6-OHDA-lesioned animals were treated for 21 days with GPS (25 or 50 mg/kg) and/or L-DOPA (20 mg/kg). Dopaminergic neuronal cell death was assessed by counting tyrosine hydroxylase (TH)-immunopositive cells in the substantia nigra and measuring levels of dopamine, norepinephrine, 3,4-dihydroxyphenylacetic acid (DOPAC), and homovanillic acid (HVA) in the striatum. Dopaminergic neuronal cell death induced by 6-OHDA lesions was ameliorated by GPS treatment (50 mg/kg). L-DOPA treatment exacerbated 6-OHDA-induced dopaminergic neuronal cell death; however, these effects were partially reversed by GPS treatment (25 and 50 mg/kg). These results suggest that GPS treatment is protective against dopaminergic neuronal cell death in a 6-OHDA-lesioned rat model of PD with long-term L-DOPA treatment. Therefore, GPS may be useful as a phytotherapeutic agent for the treatment of PD.

Effect of gypenosides on RAGE expression and oxidative stress in human mesangial cells induced by AGEs / 中草药

Objective: To observe the effect of gypenosides (GP) on the expression of receptor for advanced glycated endproducts (RAGE) and the oxidative stress status of human mesangial cells (HMCs) induced by AGEs. Methods: HMCs were cultured in DMEM of low glucose containing 13% fetal bovine serum in vitro, which were divided into six groups: normal control group (DMEM of low glucose containing 13% fetal bovine serum), AGEs group (AGEs 200mg/L), GP low-dose group (25 mg/L and AGEs 200 mg/L), GP mid-dose group (75 mg/L and AGEs 200 mg/L), GP high dose group: (150 mg/L and AGEs 200 mg/L), and aminoguanidine positive control group (0.1 mmol/L and AGEs 200 mg/L). The expression of RAGE mRNA levels of each group was detected using semi-quantitative RT-PCR method, protein expression levels by Western blotting after cultured 72 h. Simultaneously, the levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in cell supernatant, and intracellular glutathione trace (GSH) in cell were measured. Results: AGEs could significantly promote the expression of RAGE and mRNA in HMCs (P<0.01), and enhance cellular oxidative stress levels. GP could effectively inhibit the expression of RAGE and mRNA, increase the level of SOD in cell supernatant and GSH in cell, reduce the level of MDA in cell supernatant in a dose-dependent manner compared with the control group, the difference was significant (P<0.05). Conclusion: AGEs stimulation can induce the high expression of RAGE in HMCs and enhance the level of oxidative stress in vitro. GP can down the high expression of RAGE stimulated by AGEs, while improve the oxidative stress in cells, and its possible mechanism of GP may block AGEs-RAGE-oxidative stress signaling pathways mediated by RAGE, it shows the lower levels of MDA and SOD and higher levels of GSH in cell.

Research on GPs Inhibition of miRNA-122 Expression and Lipid-lowering Effect Via Regulation of Lipid Metabolism Enzyme Activity / 世界科学技术-中医药现代化

This study was aimed to explore whether gypenosides (GPs) can inhibit the expression of miRNA-122 and regulate the lipid metabolism enzyme activity to play a role in lipid-lowering effect. A total of 48 healthy male SD rats were randomly divided into 4 groups, which were the normal control group (C), hyperlipidemic model group (M), simvastatin group (S) and the GPs group (G). All groups were fed with high-fat diets except the normal control group which was fed with normal diets. The GPs, which were dissolved in 0.3% sodium carboxymethyl cellulose (CMC-Na) solution, were given by the intragastric administration. The C group and M group were given 0.3% CMC-Na solution (1 mL/100 g) daily. The G group was given 160 mg·kg-1 of GPs daily. The S group was given 5 mg·kg-1 of simvastatin daily. The experiment was continued for 8 weeks. After the last medication, rats were fasted for 12 hours. Rats were anesthetized with chloral hydrate (7%). Abdominal arterial blood samples were collected to detect the total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C). The wet weight of liver was weighed and the liver index was measured. The liver total RNA was extracted to determine the expression of miRNA-122 by the real-time PCR. The homogenates of liver tissues were prepared for the determination of hepatic lipase (HL), lipoprotein lipase (LPL) and HMG-CoA reductase activity. Cholesterol micelle formation experiments were implementedin vitro. The results showed that compared with the normal control group, TC, TG and LDL-C levels of the model group were significantly increased (P< 0.01), while the HDL-C levels in each group were obviously decreased (P< 0.05). Compared with the model group, TC, TG and LDL-C levels of the S group and G group were obviously decreased (P< 0.05), and the HDL-C level was obviously increased (P< 0.05). Compared with the model group, the liver indexes of the S group and G group were obviously decreased (P< 0.05). Compared with the hyperlipidemia model group, the expressions of miRNA-122 of the S group and G group were significantly reduced (P< 0.05). Compared with the hyperlipidemia model group, the activity of HMG-CoA reductase was obviously reduced in the S group and G group (P< 0.05), but the HL and LPL activities were obviously increased (P< 0.05). GPs can inhibit the formation of cholesterol micelles to some extent. It was concluded that GPs can effectively reduce the blood lipid level in hyperlipidemic rats, in order to relieve the hepatic fatty lesions. Its lipid-lowering mechanism was related to its inhibition of miRNA-122 expression and regulation of lipid metabolism enzyme activity, as well as the inhibition on the formation of cholesterol micelles.

Effect of Gypenosides on Lipid Peroxidation in Rats with Type 2 Diabetes Mellitus Complicated by Nonalcoholic Fatty Liver Disease / 医药导报

Objective To obserVe the effect of gyPenoside on liPid Peroxidation and hePatic lesion in rats with tyPe 2 diabetes mellitus and nonalcohol fatty liVer disease. Methods Totally,65 SPF male SD rats were randomly diVided into blank control grouP (grouP N),NAFLD model grouP (grouP NM),and NAFLD with T2DM model grouP. The NAFLD with T2DM model grouP was further diVided into three subgrouPs:JH grouP,Perfused with 1 g·kg-1 ·d-1 GPS;JL grouP,Perfused with 0. 5 g·kg-1 ·d-1 GPS;model control grouP,Perfused with the same Volume of water. Blood sugar,triglycerides ( TG) ,total cholesterol ( TC) ,alanine aminotransferase ( ALT) ,asPart aminotransferase ( AST) ,adePonectin ( ADP) in the Plasma were measured. TG, malondialdehyde (MDA),and suPeroxide dismutase (SOD) in the liVer tissue were also tested. Results ADP leVel was (7. 46±1. 12),(3. 58±0. 98),(4. 89±1. 02),(4. 79±1. 01) and (4. 13±0. 89) ng·mL-1 in N,M,NM,JH and JL grouPs, resPectiVely. The ADP leVel was significantly higher in grouP JH and JL than in grouP M (P<0. 01),and significantly higher in grouP JH than in grouP JL (P<0. 05). MDA leVel was (2. 98±0. 09),(4. 22±0. 11),(3. 66±0. 10),(3. 72±0. 11),(3. 99±0. 13) nmol·mL_1 in N,M,NM,JH and JL grouPs,resPectiVely. The MDA leVel was significantly lower in grouP JH and JL than in grouP M (P<0. 01),and significantly lower in grouP JH than in grouP JL (P<0. 05). SOD leVel was (240. 8±17. 4), (149. 9±20. 6),(181. 6±19. 4),(209. 8±19. 2),(189. 4±18. 9) U·mL_1 in N,M,NM,JH,and JL grouPs,resPectiVely. SOD leVel was significantly higher in grouP JH and JL than in grouP M (P<0. 01),and significantly higher in grouP JH than in grouP JL (P<0. 05). TG leVel was (28. 98±1. 68),(214. 46±5. 44),(198. 46±6. 98),(142. 87±6. 64) and (164. 92±7. 56) mg·g-1 in N,M,NM,JH and JL grouPs,resPectiVely. TG leVel was significantly lower in grouP JH and JL than in grouP M ( P<0. 01),and significantly lower in grouP JH than in grouP JL (P<0. 05). ALT and AST were significantly lower in grouP JH and JL than in grouP M (P<0. 01),and significantly lower in grouP JH than in grouP JL (P<0. 05). Conclusion The liPid Peroxidation in the liVer of rats with T2DM comPlicated with NAFLD can be reduced by gyPenoside,and hePatic lesion may be alleViated through inhibition of liPid Peroxidation.

Preparation and characterization of gypenosides liposomes consisted by lecithin and stigmasterol / 中草药

Objective: To study the feasibility of preparing gypenoside liposomes (GLs) with stigmasterol instead of cholesterol, and to optimize the prescription and preparation of GLs. Methods: GLs without cholesterol were prepared by ethanol injection method with lecithin and stigmasterol, and the encapsulation efficiency was determined by protamine precipitation method. The encapsulation efficiency was adopted to evaluate and optimize the preparation process as response in orthogonal design assay. GLs were characterized by Dynamic Light Scattering and Atomic Force Microscope (AFM) using particle size and electric potential as indexes. Results: Ethanol injection was the best method to prepare GLs with lecithin and stigmasterol. The optimal preparation process determined by orthogonal design assay was as follows: the ratio of gypenosides to lecithin was 1:10, the ratio of lecithin to stigmasterol was 4:1, the pH value of PBS buffer solution was 7.0, and the hydration temperature was 35°C. The entrapment efficiency was (77.00 ± 1.17)%, the particle size was (202.6 ± 3.9) nm, the PDI was 0.108 ± 0.003, the zeta potential was (-28.66 ± 0.86) mV, and the morphology was round observed by AFM. Conclusion: The preparation of GLs with lecithin and stigmasterol was feasible. The GLs prepared by optimum preparation process have high encapsulation efficiency, good morphology, and reproducibility.

The effect of gypenosides on hydrogen sulfide in liver tissue and plasma of rat with type 2 diabetes mellitus and nonalcoholic fatty liver disease / 国际中医中药杂志

Objective To observe the influence ofgypenosides on hydrogen sulfide in liver tissue and plasma of rat with type 2 diabetes mellitus and nonalcoholic fatty liver disease.Methods 58 SPF male SD rats,with body mass 220～250 g,were randomly divided into a blank control group (group N,n=7),and a NAFLD and T2DM model group (Group M,n=51).Group N was fed with ordinary diet in the first four weeks,group M was fed with diets of high fat and sugar,injected with 40 mg/kg STZ overnight,and the same diets for the next four weeks.The rat model with T2DM and NAFLD was build.NAFLD and T2DM model group were divided into three groups:a high dose GPS group (JH,n=9) injected with 1 g/kg · d-1 GPS,a low dose GPS group (JL,n=9) injected with 0.5 g/kg · d-1 GPS,and a model group (M,n=9) injected with the same volume of water,and high fat diet at the same time.The treatment period was six weeks,and the experiment period was fourteen weeks.TG,TC,BS,and H2S in the plasma of rat were tested,and H2S in the liver tissue of the rat was tested.Results ①The changes of H2S in plasma:group JH [(4.30±0.43) μmol/L] and JL [(3.83 ±0.47) μmol/L] was lower than group M [(2.67 ± 0.41) μmol/L],there was a significant difference.②The changes of H2S in the liver tissue:group JH [(333.52±37.94) pmol/min/mg/protein] and JL [(275.81 ±36.07)pmol/min/mg/protein] was lower than group M [(237.8± 33.05) pmol/min/mg/protein],there was a significant difference.③BS levels:group JH(10.86±3.46)mmol/L,group JL (14.78±3.39)mmol/L,group M(18.84±4.24) mmol/L,group JH and JL was lower than group M,there was a significant difference (P＜0.01).④The plasma TG level:group N (0.96±0.09) mmol/L,group JH (2.82± 0.66) mmol/L,group JL (1.83± 0.56) mmol/L,group M (3.97 ± 0.64) mmol/L.group JH and JL was lower than group M,there was a significant difference (P＜0.01).Conclusion Gypenoside can reduce the blood sugar,triglycerides,and total cholesterol in rat with with type 2 diabetes mellitus and nonalcohol fatty liver disease.H2S concentrations in plasma and liver tissue of the rats with T2DM and NAFLD were increased by GPS,showing dose dependence.Gypenosides can also improve metabolism of blood glucose and lipid in rats with T2DM and NAFLD.

Effects of gypenosides on the expressions of p53 and p21 protein in photodamaged BALB/c mouse skin / 中华皮肤科杂志

Objective To explore the mechanism underlying the effects of gypenosides (GP) against photodamage. Methods Eighty BALB/c mice were equally divided into 8 groups, i.e., blank control group (untreated), UVB model group (irradiated with UVB), GP I group (irradiated with UVB before topical application of GP), GPⅡ group (irradiated with UVB followed by topical application of GP), VitE I group (irradiated with UVB after topical application of Vitamine E cream), VitE Ⅱ group (irradiated with UVB followed by topical application of Vitamine E cream), Vehicle group Ⅰ (irradiated with UVB after application of the drug vehicle),and Vehicle group Ⅱ (irradiated with UVB before application of the drug vehicle). UVB irradiation was performed once every other day for 14 days. Mice were sacrificed after the last irradiation and skin specimens were obtained from the irradiated sites, and the levels of p53 and p21 protein were measured by Western blot in the specimens. Results The expression level of p53 protein was significantly lower in the blank control group than in the UVB model group (0.11 ± 0.08 vs. 0.22 ± 0.12) and GP Ⅰ group (0.44 ± 0.23, P < 0.01),in the blank control group and UVB model group than in the GP Ⅱ group (0.48 ± 0.24, P < 0.01, 0.05). VitE Ⅰ group (0.49 ± 0.29) and VitE II group (0.50 ± 0.27) were similar to the GP groups in the expression of p53 protein. No statistical difference was observed in the expression of p21 protein between the eight groups. Conclusion The upregulation of p53 protein expression in epidermal cells may be related to the mechanisms underlying the protective effect of 1.5% GP cream against photodamage.

Effects of gypenosides to the immune deficit mouse's non-specific immunological function / 中国基层医药

Objective To explore the pharmacological function of immune enhancer of gypenosides and discuss the dose-effect relationship.Methods The gypenosides was used as an immune enhancer to cure the immune deficit mouse and observe non-specific immunological function by carbon clear test.All of the mice were administrated with CTX and then were divided into six groups:control group,high dose group,middle dose group,lower dose group.Respectively,the carbon clear test was performed in each group and the variation of non-specific immune function was observed.Results The carbon clear test showed that the index of carbon clear test in control group was markedly decreased while the index of carbon clear testing high and middle group was increased.The effect of gypenosides was dose-dependent on the non-specific immune.Conclusion Gypenosides can markedly increase the non-specific immune function.