miR-188-5p对肾间质纤维化过程中MMP-13表达的影响

目的:探讨miR-188-5p对肾间质纤维化过程中基质金属蛋白酶(matrixmetalloproteinases,MMPs)表达的影响。方法:采用TGF-β1诱导肾小球系膜细胞(Mesangial cells,MC)纤维化,观察纤维化过程中miR-188-5p和MMP-13的表达变化;通过报告基因实验研究miR-188-5p对MMP-13的调控作用;通过脂质体转染的方法将人工合成的miR-188-5p mimics/inhibitor转入细胞内增加或减少miR-188-5p的表达,进一步观察miR-188-5p对MMP-13表达的影响。结果:TGF-β1诱导肾小球系膜细胞纤维化后miR-188-5p的表达明显升高,而MMP-13表达下调;报告基因实验表明miR-188-5p对MMP-13的表达有明显的调控作用,减少miR-188-5p的表达后MMP-13的表达上调,而增加miR-188-5p的表达后MMP-13的表达下调,呈明显负性调控。结论:在肾间质纤维化过程中,miR-188-5p可能通过抑制MMP-13的表达而发挥促纤维化的作用,本研究为肾间质纤维化的治疗提供了新的靶点。

Effect of miR-188-5p on the Expression of MMP-13 During RenalInterstitial Fibrosis in Mice*

Objective： To explore the role of miR- 188-5p on the expression of matrixmetallo- proteinases 13 （MMPs- 13） during renal interstitial fibrosis in mice. Methods： Mice renal mesangial cells （MC） were treated with TGF-β1 at concentration of 2ng/ml for 24h, then the mRNA and protein level of miR-188-5p and MMP-13 were tested. Next, reporter gene experiment was used to verify the effect ofmiR-188-5p on the regulation of MMP-13. In order to observe the effect on MMP-13, level ofmiR-188-5p in MC cell were then up/down regulated by mimics/inhibitor through Lipofectamine transfection. Results： TGF-β1 increased the expression of the miR-188-5p, while the mRNA and protein level of MMP-13 were reduced. Reporter gene experiment confirmed the negative regulation ofmiR-188-5p on MMP-13 expression. Up-regulation of miR-188-5p could inhibit the expression of MMP-13, while down-regulation of miR-188-5p could increade the level of MMP-13. Conclusion： In renal interstitial fibrosis, miR-188-5p may play a fibrogenic role by down-regulating the expression of MMP-13. This study provided a new therapeutic target for renal interstitial fibrosis.