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TGF-β_1诱导的肾小管上皮细胞转分化是肾小管问质纤维化的重要细胞过程。泛素-蛋白酶体系统在此过程中起重要的调控作用。通过体外观察蛋白酶体抑制剂MG-132对TGF-β_1诱导的HK-2细胞株转分化作用的影响,并探讨其分子机制。RT-PCR法检测Smurf1、Smurf2和Smad7 mRNA的表达,免疫印迹法检测Smurf1、Smurf2和Smad7蛋白的表达,免疫组织化学法检测胞浆中FN和α-SMA的分泌。结果显示:MG-132干预组细胞Smurf1、Smurf2、Smad7 mRNA表达下降,而Smad7蛋白表达增加,胞浆中FN和α-SMA分泌减少。结果表明,MG-132可通过抑制TGF-β_1诱导的HK-2Smurf1、Smurf2的表达,抑制Smad7在26S蛋白酶体中的降解,减轻FN和α-SMA的分泌和沉积,从而抑制HK 2转分化,延缓肾小管间质纤维化的发展。
Transdifferentiation of renal tubular epithelial cells induced by TGF-β_1 is an important cellular process of renal tubule fibrosis. The ubiquitin-proteasome system plays an important regulatory role in this process. The effect of proteasome inhibitor MG-132 on TGF-β1-induced transdifferentiation of HK-2 cell line was observed in vitro and its molecular mechanism was explored. The expression of Smurf1, Smurf2 and Smad7 mRNA were detected by RT-PCR. The expression of Smurf1, Smurf2 and Smad7 protein were detected by Western blotting. The secretion of FN and α-SMA in cytoplasm was detected by immunohistochemistry. The results showed that the mRNA expression of Smurf1, Smurf2 and Smad7 in MG-132 intervention group decreased, while the expression of Smad7 protein increased, while the secretion of FN and α-SMA in cytoplasm decreased. The results showed that MG-132 could inhibit HK 2 transdifferentiation, down-regulate the expression of HK-2Smurf1 and Smurf2 induced by TGF-β 1, inhibit the degradation of Smad7 in 26S proteasome, alleviate the secretion and deposition of FN and α-SMA, Delay the development of tubulointerstitial fibrosis.