马里苷通过AMPK通路抑制高糖软脂酸诱导大鼠肾小球系膜细胞氧化应激及炎症损伤的作用机制

Marein inhibited oxidative stress and inflammation of high glucose induced HBZY-1 cells via AMPK signaling pathway

目的 研究马里苷介导AMPK信号通路,延缓糖尿病肾病(DN)氧化应激、炎症损伤,抑制纤维蛋白表达的作用。方法高糖软脂酸诱导大鼠肾小球系膜细胞(HBZY^-1)建立体外DN模型,将细胞分为:正常对照组(NG)、模型组(葡萄糖100 mmol·L ^-1 +软脂酸250 μmol·L ^-1 ,HG+PA)、马里苷不同剂量组(25、50、100、200 μmol·L ^-1)。采用MTS法检测马里苷对HBZY^-1细胞增殖的影响,从中筛选最适给药浓度;Western blot法检测各组细胞中NOX4、MCP^-1、TGF-β1、FN、α-SMA、Collagen Ⅵ,以及AMPK蛋白家族中AMPKγ1、p-AMPKα的表达水平。结果马里苷抑制高糖软脂酸诱导的HBZY^-1细胞增殖;下调模型细胞中NOX4、TGF-β1、MCP^-1、α-SMA、FN及Collagen Ⅵ的表达;上调p-AMPKα及AMPKγ1的表达。结论马里苷可能通过活化AMPKγ1,激活AMPK信号通路,抑制HBZY^-1细胞氧化应激、炎症反应与纤维化蛋白的表达,延缓DN肾脏损伤。

Aim To study the role of marein mediated AMPK signaling pathway in delaying oxidative stress,inflammation and fibrotic protein expression in diabetic nephropathy(DN).Methods In vitro diabetic nephropathy model was established by HG+PA induced rat mesangial cells(HBZY-1),and the cultured HBZY-1 cells were divided into normal control group(NG),HG+PA(Glucose100 mmol·L^-1 +Palmitic acid 250 μmol·L^-1 ,HG+PA) model group,HG+PA+marein with different doses of 25 μmol·L^-1 ,50 μmol·L^-1 ,100 μmol·L^-1 ,and 200 μmol·L^-1 groups.MTS was used to detect the effect of marein on HBZY-1 cell proliferation,and the optimal concentration was selected.Western blot was used to test the protein expression of NOX4,TGF-β1,MCP-1,α-SMA,FN,Collagen Ⅵ.Adenosine monophosphate activated(AMPK) protein kinase family of AMPKγ1,p-AMPK α expression were measured.Results Marein inhibited high glucose palmitate-induced proliferation of HBZY-1 cells,down-regulated NOX4,TGF-β1,MCP-1,α-SMA,FN and Collagen Ⅵ expression in model cells.Meanwhile,marein up-regulated both AMPK γ1 and p-AMPKα expression.Conclusions Marein may inhibit the HBZY-1 cell proliferation,oxidative stress,inflammation and fibrosis factors expression in HG+PA induced HBZY-1 cell by activating of both AMPK γ1 and AMPK signaling pathway,thus delaying renal injury in diabetic nephropathy.