达格列净拮抗糖尿病肾脏纤维化作用研究

Study on the Antagonistic Effect of Dapagliflozin on Diabetic Renal Fibrosis

目的探讨达格列净对糖尿病小鼠肾脏以及棕榈酸诱导的小鼠肾小球系膜细胞(SV40)的保护作用。方法将36只SPF级C57BL/6小鼠随机分为正常组、糖尿病组、达格列净组,体外培养SV40并分为正常组、棕榈酸组、正常对照组、棕榈酸+达格列净组。Masson染色检测小鼠肾脏组织中肌纤维与胶原纤维表达情况,Western blot检测小鼠肾脏组织与SV40中Ⅰ型胶原、Ⅲ型胶原和转化生长因子β1(TGF-β1)蛋白表达,qPCR检测小鼠肾脏组织与SV40中Ⅰ型胶原、Ⅲ型胶原、α-平滑肌肌动蛋白(α-SMA)mRNA含量变化,CCK-8法检测SV40增殖水平。结果达格列净可明显改善糖尿病小鼠肾脏纤维化,抑制SV40细胞增殖,降低糖尿病小鼠肾脏组织与SV40中Ⅰ型胶原、Ⅲ型胶原、TGF-β1与α-SMA表达。结论达格列净通过减少胶原沉积,下调TGF-β1与α-SMA,发挥对糖尿病肾脏纤维化的保护机制。

Objective To investigate the protective effect of Dapagliflozin on diabetic mouse kidneys and palmitic acid-induced mouse glomerular mesangial SV40 cells.Methods Thirty-six SPF-grade C57BL/6 mice were randomly divided into normal group,diabetes group,and Dapagliflozin group.SV40 cells were cultured in vitro and divided into normal group,palmitic acid group,and palmitic acid+Dapagliflozin group.Masson staining was used to detect the expression of myofibrils and collagen fibrils in mouse kidney tissues.Western blot was used to detect the protein expression of collagenⅠ,collagenⅢand TGF-β1 in mouse kidney tissues and SV40 cells.qPCR was used to detect the mRNA level changes of collagenⅠ,collagenⅢandα-SMA in mouse kidney tissues and SV40 cells.CCK-8 method was used to detect the proliferation level of SV40 cells.Results Dapagliflozin significantly improved renal fibrosis in diabetic mice,inhibited SV40 cell proliferation,and reduced the expression of collagenⅠ,collagenⅢ,TGF-β1 andα-SMA in kidney tissues of diabetic mice and SV40 cells.Conclusion Dapagliflozin can exert a protective mechanism against diabetic renal fibrosis by reducing collagen deposition and down-regulating TGF-β1 andα-SMA expression.