microRNA-132对肾间质成纤维细胞分泌细胞外基质的调控作用

Effect of adjusting and controlling microRNA-132 on extracellular matrix of secretion cells in renal interstitial fibroblast

目的:探讨microRNA-132(miR-132)对转化生长因子(transforming growth factor,TGF)β1刺激后大鼠肾间质成纤维细胞(NRK-49F)的细胞外基质分泌的调控作用。方法:体外培养NRK-49F细胞株,经TGF-β1(2、3、5 ng/mL)作用24 h,或细胞转染miR-132模拟物(rno-miR-132 mimics,50 ng/mL)作用24 h,或rno-miR-132 mimics与TGF-β1共同作用24 h,茎环实时荧光定量PCR(real-time quanti-tative PCR,RT-qPCR)法检测细胞miR-132表达,RT-qPCR法和Western Blot法分别检测细胞胶原蛋白I(Collagen I,Col I)mRNA和蛋白表达。结果:①不同浓度TGF-β1刺激后NRK-49F细胞miR-132和Col I的表达显著升高(P<0.01)。②rno-miR-132 mimics(50 ng/mL)作用NRK-49F细胞Col I表达显著升高(P<0.01)。③与TGF-β1(3 ng/mL)阳性对照组相比,rno-miR-132 mimics(50 ng/mL)与TGF-β1(3 ng/mL)共同作用NRK-49F细胞Col I表达显著升高(P<0.01)。结论:miR-132可以促进TGF-β1诱导的NRK-49F细胞分泌细胞外基质。

Objective： To explore the adjusting and controlling effect of microRNA-132 （miR-132） on extracellular matrix of the secretion cells in renal interstitial fibroblast （NRK-49F） cells after stimulation of TGF-151. Methods： The cultured NRK-49F cells were treated with different concentration （2,3,5 ng/mL） TGF-01 alone, or with rno-miR-132 mimic （50 ng/mL） alone, or co-treated with rno-miR-132 mimics and TGF-β1 for 24 hours. Stem ring RT-qPCR was used to assay the expression of miR- 132. The expressions of mRNA and protein of Collagen I （Col I） were assessed with RT-qPCR and western blot respectively. Results： ①miR-132 and Col I expressions increased in a dose-dependent manner （P〈0.01） treated with TGF- β1. ② Treated with mo-miR-132 mimics, mRNA and protein of Col I expression increased （P〈0.01）. ③Compared with TGF- β1 treated group, the expression of Col I treated with rno-miR-132 mimics and TGF-β1 both groups was increased （P〈O.O1）. Conclusion： MiR-132 can promote the TGF- β1inducing secretion and deposition of extracellular matrix.