| 摘要: |
| 目的：观察参苓白术散对非酒精性脂肪性肝病（NAFLD）大鼠肝细胞、Kupffer细胞、哺乳动物雷帕霉素靶蛋白（mTOR）通路相关基因及蛋白表达的影响,以阐明参苓白术散抗NAFLD的作用机制。方法：将80只大鼠随机分为正常组,模型组及参苓白术散低、高剂量组。对模型组、各药物组连续8周喂养高脂饲料制备NAFLD大鼠模型。造模成功后采用油红O染色观察肝组织病理变化;Ⅳ型胶原酶离体循环灌注法分离肝细胞、Kupfer细胞;全自动生化仪检测血清总胆固醇（TC）、三酰甘油（TG）;酶联免疫吸附法测定肝细胞及Kupffer细胞血管内皮生长因子（VEGF）、干扰素（IFN）-γ含量;实时荧光定量PCR及蛋白质印迹法（Western blot）检测大鼠肝细胞及Kupffer细胞脑内富含的小G蛋白Ras同系物（Rheb）、mTOR复合物1（mTORC1）、核糖体S6蛋白激酶1（S6K1）和真核细胞翻译起始因子4E-结合蛋白1（4E-BP1）mRNA和蛋白表达。结果：模型组大鼠肝组织存在严重的脂质蓄积。血清TC、TG,肝细胞、Kupffer细胞,VEGF、IFN-γ含量,以及Rheb、mTORC1、S6K1、4E-BP1 mRNA和蛋白表达水平较正常组显著升高（P<0.01）,说明造模成功。与模型组比较,各药物干预组血清TC、TG,肝细胞、Kupffer细胞,VEGF、IFN-γ含量,以及Rheb、mTORC1、S6K1、4E-BP1 mRNA和蛋白表达均有不同程度的下调（P<0.05或P<0.01）。其中参苓白术散高剂量组下调趋势最为显著,与低剂量组比较,差异均有统计学意义（P<0.05或P<0.01）。结论：参苓白术散抗NAFLD的作用机制可能与抑制肝细胞及Kupffer细胞mTOR通路中Rheb、mTORC1、S6K1、4E-BP1基因及蛋白表达有关。 |
| 关键词: 非酒精性脂肪性肝病 参苓白术散 mTOR通路 肝细胞 Kupffer细胞 |
| DOI： |
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| Effect of Shenling Baizhu powder on hepatocytes,Kupffer cells,and expression of genes and proteins associated with the mammalian target of rapamycin pathway in rats with nonalcoholic fatty liver disease |
| XU Yongjian,ZHANG Yuncheng,FENG Gaofei |
| (Shenzhen Hospital of Beijing University of Chinese Medicine（Longgang）,Shenzhen 518172,Guangdong,China) |
| Abstract: |
| Objective：To investigate the effect of Shenling Baizhu powder on hepatocytes,Kupffer cells,and expression of genes and proteins associated with the mammalian target of rapamycin (mTOR) pathway in rats with nonalcoholic fatty liver disease (NAFLD),and to clarify the anti-NAFLD mechanism of Shenling Baizhu powder.Methods：A total of 80 rats were randomly divided into normal group,model group,and low- and high-dose Shenling Baizhu powder groups.The rats in the model group and the low- and high-dose Shenling Baizhu powder groups were given high-fat diet to establish a rat model of NAFLD.After successful modeling,oil red O staining was used to observe liver pathological changes;in vitro circulated perfusion with collagenase type IV was performed to isolate hepatocytes and Kupffer cells;an automatic biochemical analyzer was used to measure the serum levels of total cholesterol (TC) and triglyceride (TG);ELISA was used to measure the content of vascular endothelial growth factor (VEGF) and interferon-γ (IFN-γ) in hepatocytes and Kupffer cells;quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression of Ras homolog enriched in brain (Rheb),mTOR complex 1 (mTORC1),S6 kinase 1 (S6K1),and eukaryotic translation initiation factor 4E binding protein 1 (4E-BP1) in hepatocytes and Kupffer cells.Results：Severe lipid accumulation was observed in the rats in the model group.Compared with the rats in the normal group,the rats in the other groups had significantly increases in serum TC and TG,content of VEGF and IFN-γ in hepatocytes and Kupffer cells,and mRNA and protein expression of Rheb,mTORC1,S6K1,and 4E-BP1 in hepatocytes and Kupffer cells (P＜0.05),suggesting that the model was established successfully.Compared with the model group,the drug intervention groups had varying degrees of reductions in serum TC and TG,content of VEGF and IFN-γ in hepatocytes and Kupffer cells,and mRNA and protein expression of Rheb,mTORC1,S6K1,and 4E-BP1 in hepatocytes and Kupffer cells (P＜0.05 or P＜0.01),among which the high-dose Shenling Baizhu powder group had the greatest reductions,and there were significant differences between the high- and low-dose Shenling Baizhu powder groups (P＜0.05 or P＜0.01).Conclusion：Shenling Baizhu powder can exer t an anti-NAFLD effect by inhibiting the mRNA and protein expression of Rheb,mTORC1,S6K1,and 4E-BP1 associated with the mTOR pathway in hepatocytes and Kupffer cells. |
| Key words: nonalcoholic fatty liver disease Shenling Baizhu powder mammalian target of rapamycin pathway hepatocyte Kupffer cell |
