ABSTRACT
Objective: The protective effect of Xiaochaihu Decoction on non-alcoholic steatohepatitis (NASH) model mice was studied by constructing a methionine-choline deficiency (MCD) diet-induced NASH model in mice. Methods: C57BL/6 mice, as the research objects, were randomly divided into normal control group, model group, Xiaochaihu Decoction (high, medium and low doses) group, Yishanfu group and Qianggan Capsule group. The NASH model was established by feeding MCD feeds, and model intervention was carried out at the same time by giving different drugs in groups; Changes in body weight, daily food intake, and daily water volume of the mice were recorded during the experiment. HE staining of liver tissue was performed at the end of the experiment to observe pathological changes. and the levels of biochemical indicator of alanine aminotransferase (ALT), Aspartate aminotransferase (AST), total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), interleukin 6 (IL-6), and tumor necrosis factor-α (TNF-α) in serum, and the changes of TC and TG levels in liver tissue were detected, RT-PCR was used to detect the expression levels of fatty acid synthase (FAS) and sterol regulatory element binding protein 1c (SREBP-1c) in liver tissues. Results: Data such as mouse weight, daily food intake, daily water intake, and organ coefficients indicated that MCD diet-induced model mice showed weight loss, decreased intake, and decreased liver wet weight. The weight, intake and liver coefficient of mice in Xiaochaihu Decoction group were significantly higher than those in the model group; The results of HE staining showed that Xiaochaihu Decoction could significantly reduce the degree of steatosis and inflammation of liver tissue, and improve the morphology and structure of liver cells; The results of serum biochemical indicators showed that Xiaochaihu Decoction significantly reduced the levels of TG, TC, AST, ALT, IL-6, TNF-α and increased the level of HDL-C in NASH model mice; RT-PCR results showed that the gene expression levels of FAS and SREBP-1c in the liver tissue of the model group mice were significantly increased, and the administration of Xiaochaihu Decoction could significantly reduce the gene expression levels of FAS and SREBP-1c. Conclusion: Xiaochaihu Decoction has obvious protective effect on NASH mouse model induced by MCD diet. It may play a lipid-lowering role by regulating the expression of inhibitors of fatty acid synthetase genes (FAS, SREBP-1c), reducing fat accumulation, and inhibiting the expression of inflammatory factors to improve liver tissue damage.
ABSTRACT
Farnesoid X receptor (FXR) is a member of the nuclear receptor superfamily of ligand-activated transcription factors. As a metabolic regulator, FXR plays key roles in bile acid and cholesterol metabolism and lipid and glucose homeostasis. Therefore, FXR is a potential drug target for several metabolic syndromes, especially those related to lipidemia disorders. In the present study, we identified small molecule SIPI-7623, a derivative of an extract from Oriental wormwood (Artemisia capillaris), and found that it specifically upregulated the expression of cholesterol-7-alpha-hydroxylase (CYP7A1), downregulated the expression of sterol-regulatory element-binding protein 1c (SREBP-1c) in the liver, and inhibited the expression of ileal bile acid binding-protein (IBABP) in the ileum of rats. We found that inhibition of FXR by SIPI-7623 decreased the level of cholesterol and triglyceride. SIPI-7623 reduced the levels of cholesterol and triglyceride in in vitro HepG2 cell models, ameliorated diet-induced atherosclerosis, and decreased the serum lipid content on rats and rabbits model of atherosclerosis in vivo. Furthermore, SIPI-7623 decreased the extent of atherosclerotic lesions. Our resutls demonstrated that antagonism of the FXR pathway can be employed as a therapeutic strategy to treat metabolic diseases such as hyperlipidemia and atherosclerosis. In conclusion, SIPI-7623 could be a promising lead compound for development of drugs to treat hyperlipidemia and atherosclerosis.
Subject(s)
Animals , Humans , Male , Rabbits , Rats , Artemisia , Chemistry , Atherosclerosis , Drug Therapy , Genetics , Metabolism , Cholesterol , Metabolism , Cholesterol 7-alpha-Hydroxylase , Genetics , Metabolism , Drugs, Chinese Herbal , Hyperlipidemias , Drug Therapy , Genetics , Metabolism , Hypolipidemic Agents , Liver , Metabolism , Rats, Sprague-Dawley , Receptors, Cytoplasmic and Nuclear , Genetics , Metabolism , Sterol Regulatory Element Binding Protein 1 , Genetics , Metabolism , Triglycerides , MetabolismABSTRACT
Farnesoid X receptor (FXR) is a member of the nuclear receptor superfamily of ligand-activated transcription factors. As a metabolic regulator, FXR plays key roles in bile acid and cholesterol metabolism and lipid and glucose homeostasis. Therefore, FXR is a potential drug target for several metabolic syndromes, especially those related to lipidemia disorders. In the present study, we identified small molecule SIPI-7623, a derivative of an extract from Oriental wormwood (Artemisia capillaris), and found that it specifically upregulated the expression of cholesterol-7-alpha-hydroxylase (CYP7A1), downregulated the expression of sterol-regulatory element-binding protein 1c (SREBP-1c) in the liver, and inhibited the expression of ileal bile acid binding-protein (IBABP) in the ileum of rats. We found that inhibition of FXR by SIPI-7623 decreased the level of cholesterol and triglyceride. SIPI-7623 reduced the levels of cholesterol and triglyceride in in vitro HepG2 cell models, ameliorated diet-induced atherosclerosis, and decreased the serum lipid content on rats and rabbits model of atherosclerosis in vivo. Furthermore, SIPI-7623 decreased the extent of atherosclerotic lesions. Our resutls demonstrated that antagonism of the FXR pathway can be employed as a therapeutic strategy to treat metabolic diseases such as hyperlipidemia and atherosclerosis. In conclusion, SIPI-7623 could be a promising lead compound for development of drugs to treat hyperlipidemia and atherosclerosis.
Subject(s)
Animals , Humans , Male , Rabbits , Rats , Artemisia , Chemistry , Atherosclerosis , Drug Therapy , Genetics , Metabolism , Cholesterol , Metabolism , Cholesterol 7-alpha-Hydroxylase , Genetics , Metabolism , Drugs, Chinese Herbal , Hyperlipidemias , Drug Therapy , Genetics , Metabolism , Hypolipidemic Agents , Liver , Metabolism , Rats, Sprague-Dawley , Receptors, Cytoplasmic and Nuclear , Genetics , Metabolism , Sterol Regulatory Element Binding Protein 1 , Genetics , Metabolism , Triglycerides , MetabolismABSTRACT
Objective To investigate the effect and mechanism of liver X receptor ( LXR ) agonist on expression of fatty acid synthase( FAS) in diabetic kidney. Methods In the part of in vivo study, immunostaining was used to detect the FAS protein expression in kidney. 16-week-old male db/db mice on C57BL/6 background were administered via gavage a LXR synthetic agonist, TO901317, at a dose of 3 mg · kg-1 · d-1 or vehicle ( 0. 5%Carboxymethyl Cellulose Sodium, CMC-Na) alone for 7 d;Quantitative RT-PCR and Western blot were used to detect mRNA and protein levels of FAS and SREBP-1. In the part of in vitro study, MCT cell(a mouse murine proximal tubule cell line)was treated with 10μmol/L TO901317 for 24 h or transfected with active SREBP-1c expression vector (SREBP-1cN). HEK293 cells(a human renal tubule cell line)were transfected with mFAS-(1. 7 kb)-luc, LXR expression vector or SREBP-1cN for 12 h. Quantitative RT-PCR and luciferase reproter assay were utilized to examine FAS mRNA level and FAS promoter activity. Results FAS was abundantly expressed in renal cortex, with low expresson in renal glomeruli. The mRNA and protein expressious of FAS in kidney of db/db mice were lowered compared with db/m mice. TO90137 treatment increased FAS mRNA expression by 1. 3-fold. TO901317 increased expression of SREBP-1 in kidneys of db/m and db/db mice by 5. 1-fold and 17-fold, respectively. TO901317 and overexpression of SREBP-1c increased expression of FAS in MCT cells by 1. 5-fold and 1. 8-fold. Transcription activity of FAS were induced by TO901317, LXR, and SREBP-1cN overexpressions in HEK293 cells. Conclusions Both direct(LXRE)and indirect(SREBP-1c)mechanisms may contribute to the up-regulation of FAS expression by LXR in renal proximal tubule cells.
ABSTRACT
Objective To observe the fatty deposition in thapsigargin-induced endoplasmic reticulum stress model in hepatocytes and to discuss the possible mechanism. Methods Hepatocytes (L02 cell and HepG2 cell line) were divided into control group and experimental group (treated with 1 μmol/L thapsigargin). Fatty deposition in the hepatocytes was observed by biochemical assay and oil red O staining. Real-time PCR was used to test the expression of SREBP-1c and LXRs mRNA. And Western blotting analysis was used to examine the expression of protein of GRP78, SREBP-1c and LXRs. Results Western blotting analysis showed that GRP78 protein expression in the experimental group was remarkably higher than that inthe control group (P<0. 05), indicating the successful establishment of the endoplasmic reticulum stress model in hepatocytes. The hepatocyte fatty deposition in the experimental group was significantly more than that in the control group 48 h after thapsigargin exposure(P<0. 01). The expression of SREBP-1c mRNA and protein in the experimental group was significantly higher than that in the control group (P<0. 05), and the expression of LXRs mRNA and proteinwas not significantly between the two groups. Conclusion Endoplasmic reticulum stress may induce hepatocyte fatty deposition throuth up-regulating SREBP-1c, and LXRs is not involved in the process.
ABSTRACT
Objective To observe the fatty deposition in thapsigargin-induced endoplasmic reticulum stress model in hepatocytes and to discuss the possible mechanism. Methods Hepatocytes (L02 cell and HepG2 cell line) were divided into control group and experimental group (treated with 1 μmol/L thapsigargin). Fatty deposition in the hepatocytes was observed by biochemical assay and oil red O staining. Real-time PCR was used to test the expression of SREBP-1c and LXRs mRNA. And Western blotting analysis was used to examine the expression of protein of GRP78, SREBP-1c and LXRs. Results Western blotting analysis showed that GRP78 protein expression in the experimental group was remarkably higher than that inthe control group (P<0. 05), indicating the successful establishment of the endoplasmic reticulum stress model in hepatocytes. The hepatocyte fatty deposition in the experimental group was significantly more than that in the control group 48 h after thapsigargin exposure(P<0. 01). The expression of SREBP-1c mRNA and protein in the experimental group was significantly higher than that in the control group (P<0. 05), and the expression of LXRs mRNA and proteinwas not significantly between the two groups. Conclusion Endoplasmic reticulum stress may induce hepatocyte fatty deposition throuth up-regulating SREBP-1c, and LXRs is not involved in the process.
ABSTRACT
Objective To explore the expression of sterol regulatory element binding protein 1C (SREBP-1C) and glucose-regulated protein 94(GRP-94)in hyperhomocysteinmia and to evaluate the effects of endoplasmic reticulum stress proteins on hepatocytes lipids metabolism. Methods After hyperhomocysteinmia C57BL/6 mice model being induced by high methionine diet, TGE and CHO of Hepatocytes were determined, and the expression of SREBP-1C and GRP-94 was assessed by RT-PCR and Western blot. All data were compared to those in control group′s. Results The level of plasmic homocysteine(Hcy) and hepatocytes TGE or CHO of high methionine diet mice at different time point significantly ascended(P
ABSTRACT
Objective To explore the effects of TNF alpha on the expression of sterol-regulatory element binding protein-1c, SREBP-1c mRNA and the content of triglyceride (TG) in the models of cultured steatosis hepatocytes. Methods Steatosis models of hepatocytes were established by adding oleic acid to the growing L-02 cells, and then cultured in present with TNF alpha or its antibody. The expressions of SREBP-1c and fatty acid synthetase (FAS) mRNA were measured with RT-PCR, lipid droplets in the hepatocytes were observed with oil red staining and the TG contents in hepatocytes were measured with analyzed kit. Results SREBP-1c mRNA was upregulated in the TNF alpha treatment group in comparison with the normal control group(P