Protective effect of the Nox4/TGF-β1/p-Smad pathway regulated by sodium tanshinone ⅡA sulfonate on Human Glomerular Mesangial Cells damage induced by high glucose
Objective:To investigate the protective effect and mechanism of sodium tanshinone IIA sulfonate(STS)on Human Glomerular Mesangial Cells(HGMC)damage induced by high glucose.Methods:Cultured HGMC were used as the research objects.The objects were divided into three groups:NG group(normal glucose,5.5 mmol/L glucose),NG+Mnt group(osmotic pressure regulation,5.5 mmol/L glucose+24.5 mmol/L mannitol)and HG group(high glucose:30 mmol/L glucose;HG+low dose of STS:2.5 μg/ml;HG+medium dose of STS:5 μg/ml;HG+high dose of STS:10 μg/ml).HGMC of each group were cultured for 48 h,CCK-8 was used to detect the viability of HGMC,and EDU-488 was used to detect the proliferation of HGMC in each group.The 2,7-dichlorofluorescein diacetate(DCFH-DA)was used to detect ROS content in each group.SOD level,MDA level and T-AOC were detected by WST-8,colorimetric method and ABTS,respectively.The contents of TGF-β1,FN and Collagen Ⅳ of intracellular fibrosis were detected by ELISA.Western blot was used to detect Nox4,TGF-β1,p-Smad 3 and Collagen Ⅳ protein expression levels in each group.Results:The proliferation rate of HGMC,the contents of ROS,MDA,TGF-β1,Collagen Ⅳ and FN,and the expression levels of Colla-gen Ⅳ,TGF-β1,p-Smad3 and Nox4 in HG group were significantly higher than those in NG group,NG+Mnt group and HG+each dose of STS groups.The differences were statistically significant(P<0.05,P<0.01).The SOD level and T-AOC in HGMC in HG group were significantly lower than those in NG group,NG+Mnt group and HG+each dose of STS groups,and the differences were statistically significant(P<0.05,P<0.01).Compared with HG+low dose of STS group,Collagen Ⅳ level in HG+medium dose of STS group was significantly decreased,while Collagen Ⅳ,FN and P-SMad3 levels in HG+high dose of STS group were significantly decreased,with statistical significance(P<0.05,P<0.01).Compared with HG+medium dose of STS group,FN level in HG+high dose of STS group was significantly decreased,and the difference was sta-tistically significant(P<0.05).Conclusion:STS alleviates high glucose-induced oxidative stress and extracellular matrix deposition in HGMC through Nox4/TGF-β1/p-Smad pathway.
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