TFEB通路的大豆苷抑制高糖诱导的肾小球系膜增殖作用机制

The mechanism of daidzin inhibiting hyperglycemic mesangial proliferation based on PPAR⁃α/TFEB pathway

〔目的〕考察大豆苷在高糖条件下对肾小球系膜细胞增殖作用的影响,并探讨其可能作用机制。〔方法〕MTT法检测肾小球系膜细胞增殖水平;荧光素酶报告基因法评估大豆苷对PPAR⁃α、TFEB转录活性影响;Q⁃PCR法检测TGF⁃β1、collagenⅣ、TFEB、LIMP2的mRNA表达量;Docking分析大豆苷和PPAR⁃α⁃LBD的结合情况。〔结果〕10、20μmol/L大豆苷组显著抑制高糖刺激下肾小球系膜细胞增殖,同时5、10、20μmol/L大豆苷组显著降低高糖条件下TGF⁃β1、colla⁃genⅣ的mRNA表达量;相比空白对照组,大豆苷能够促进PPAR⁃α、TFEB荧光素酶活力;PPAR⁃α的选择性抑制剂(GW6471)逆转了大豆苷TFEB转录活性的增加;大豆苷增加TFEB、LIMP2的mRNA的表达量,大豆苷在PPAR⁃α⁃LBD的THR279、CYS⁃278、GLU282三个氨基酸位点处与其形成稳定的氢键。〔结论〕大豆苷能够抑制高糖条件下肾小球系膜的增殖,其机制可能与结合PPAR⁃α⁃LBD调控TFEB有关。

[Objective]To investigate the effect of daidzein on the proliferation of glomerular mesangial under high glucosemedium,and explore its possible mechanism.[Methods]MTT assay was used to delect the proliferation;luciferase reportergene assay was used to evaluate transcriptional activities of PPAR-α and TFEB;the mRNA expression levels of TGF-β1,collagenⅣ,TFEB and LIMP2 were detected by Q-PCR;Autodocking analyzes daidzein binding with PPAR-α-LBD.[Results]Daidzeinsignificantly inhibited proliferation of glomerular mesangial and decreased the mRNA expression levels of TGFβ1 and Collagen Nunder high glucose medium.Compared with the blank control group,the activity of PPAR-α and TFEB luciferase was increased,but selective imhibitors of PPAR-αreversed the inrease i TFEB transcriptional activity induced by daidain.Daidain increasedthe mRNA expression of TFEB and LIMP2,and formed stable hydrogen bond wih PPAR-α-LBD at THR279,CYS-278,GLU282.[Concltusion]Daidzein imhibits glomerular mesangial proliferation under high glucose medium,and its mechanismmay be binding with PPAR-α-LBD to regulate TFEB.