马里苷活化SIRT3蛋白延缓高糖及软脂酸诱导HBZY-1细胞纤维化损伤的研究  

Research on activating SIRT3 protein by marein to delay fibrosis damage induced by high glucose and palmitic acid in HBZY-1 cells

作　　者：阿米拉·阿不拉提 郭凤 马洪艳[1] 毛新民[1] 姚蓝[1] AMILA·Abulati;GUO Feng;MA Hongyan;MAO Xinmin;YAO Lan(Institute of Traditional Chinese Medicine,Xinjiang Medical University,Urumqi 830011,Xinjiang,China;Provincial and Ministry Co-constructed State Key Laboratory Cultivation Base of Xinjiang Indigenous Medicinal Resources Utilization,CAS Key Laboratory of Chemistry of Plant Resources in Arid Regions,Xinjiang Technical Institute of Physics and Chemistry,Chinese Academy of Sciences,Urumqi 830011,Xinjiang,China)

机构地区：[1]新疆医科大学中医学院,新疆乌鲁木齐830011 [2]新疆特有药用资源利用省部共建国家重点实验室培育基地,中国科学院干旱区植物资源化学重点实验室,中国科学院新疆理化技术研究所,新疆乌鲁木齐830011

出　　处：《上海中医药杂志》2022年第9期82-88,共7页Shanghai Journal of Traditional Chinese Medicine

基　　金：新疆维吾尔自治区教育厅高校科研计划项目(XJEDU2020Y026)

摘　　要：目的研究马里苷介导沉默信息调节因子3(SIRT3)/固醇调节元件结合蛋白-1(SREBP-1)信号通路修复糖尿病肾病(DN)肾脏纤维化的作用。方法高糖(HG)和软脂酸(PA)诱导大鼠肾小球系膜细胞(HBZY-1)建立体外条件下DN模型,将细胞分为正常对照组、模型组和马里苷不同剂量组;倒置显微镜观察各组细胞数量及形态的变化;Western blot法检测各组细胞中SIRT3/SREBP-1通路蛋白、纤维化损伤相关蛋白转化生长因子-β1(TGF-β1)、磷酸化(p)-Smad同源物2(Smad2)、p-Smad3、Smad4的表达水平;采用Discovery studio软件将马里苷与SIRT3进行分子对接;免疫荧光法检测各组细胞中纤连蛋白(FN)和胶原Ⅳ(CollagenⅣ)的表达和定位。结果马里苷可抑制HG及PA诱导的HBZY-1细胞增殖,使细胞间隙增大,细胞密度降低;马里苷能下调模型细胞中SREBP-1、TGF-β1、p-Smad2、p-Smad3、Smad4蛋白的表达;马里苷能上调活性SIRT3表达,同时分子对接结果表明马里苷与SIRT3蛋白能较好结合;马里苷可抑制HG及PA诱导的细胞中FN和CollagenⅣ蛋白表达。结论马里苷可能通过活化SIRT3蛋白,介导SIRT3/SREBP-1通路,下调TGF-β1/Smads信号通路蛋白,进而减轻HG以及PA诱导HBZY-1细胞的肾脏炎症和纤维化损伤。Objective To study the role of marein in mediating SIRT3/SREBP-1 signaling pathway to repair renal fibrosis in diabetic nephropathy(DN).Methods Rats’glomerular mesangial cells(HBZY-1)were induced by high glucose(HG)and palmitic acid(PA)to establish a DN model in vitro.The cells were divided into normal control group,model group and different dosage groups of marein.Inverted microscope was used to observe the changes in the number and morphology of cells in each group.Western blot was used to detect the expression levels of SIRT3/SREBP-1 pathway protein,fibrosis damage-related proteins transforming growth factor-β1(TGF-β1),p-Smad2,p-Smad3 and Smad4 in each group.Discovery studio was used to molecularly dock marein with silent information regulator 3(SIRT3).Immunofluorescence method was used to detect the expression and localization of fibronectin(FN)and CollagenⅣof cells in each group.Results Marein inhibited the proliferation of HBZY-1 cells induced by HG and PA,which increased the intercellular and decreased the cell density.Marein down-regulated the expression of SREBP-1,TGF-β1,p-Smad2,pSmad3 and Smad4 proteins in model cells.Marein up-regulated the expression of active SIRT3,and the molecular docking results showed that marein and SIRT3 protein binded well.Marein inhibited the expression of FN and CollagenⅣprotein in cells induced by HG and PA.Conclusion Marein may reduce the kidney inflammation and fibrosis damage induced by HG and PA in HBZY-1 cells by activating SIRT3 protein,mediating the SIRT3/SREBP-1 pathway and down-regulating the TGF-β1/Smads signaling pathway protein.

关 键 词：糖尿病肾病 肾脏纤维化 马里苷 两色金鸡菊 药理作用 中药研究 

分 类 号：R285.5[医药卫生—中药学]