Department of Urology, Shenzhen Children's Hospital, Shenzhen, Guangdong Province, China.
Laboratory of Pelvic Floor Muscle Function, Shenzhen Children's Hospital, Shenzhen, Guangdong Province, China.
Braz J Med Biol Res. 2021 Jan 25;54(3):e9206. doi: 10.1590/1414-431X20209206. eCollection 2021.
Renal fibrosis is one of the most significant pathological changes after ureteral obstruction. Transforming growth factor-β (TGF-β) signaling pathway plays essential roles in kidney fibrosis regulation. The aims of the present study were to investigate effects of microRNA-302b (miR-302b) on renal fibrosis, and interaction between miR-302b and TGF-β signaling pathway in murine unilateral ureteral obstruction (UUO) model. Microarray dataset GSE42716 was downloaded by retrieving Gene Expression Omnibus database. In accordance with bioinformatics analysis results, miR-302b was significantly down-regulated in UUO mouse kidney tissue and TGF-β1-treated HK-2 cells. Masson's trichrome staining showed that miR-302b mimics decreased renal fibrosis induced by UUO. The increased mRNA expression of collagen I and α-smooth muscle actin (α-SMA) and decreased expression of E-cadherin were reversed by miR-302b mimics. In addition, miR-302b up-regulation also inhibited TGF-β1-induced epithelial mesenchymal transition (EMT) of HK-2 cells by restoring E-cadherin expression and decreasing α-SMA expression. miR-302b mimics suppressed both luciferase activity and protein expression of TGF-βR2. However, miR-302b inhibitor increased TGF-βR2 luciferase activity and protein expression. Meanwhile, miR-302b mimics inhibited TGF-βR2 mRNA expression and decreased Smad2 and Smad3 phosphorylation in vivo and in vitro. Furthermore, over-expression of TGF-βR2 restored the miR-302b-induced decrease of collagen I and α-SMA expression. In conclusion, this study demonstrated that miR-302b attenuated renal fibrosis by targeting TGF-βR2 to suppress TGF-β/Smad signaling activation. Our findings showed that elevating renal miR-302b levels may be a novel therapeutic strategy for preventing renal fibrosis.
肾纤维化是输尿管梗阻后最显著的病理变化之一。转化生长因子-β(TGF-β)信号通路在肾脏纤维化调控中发挥着重要作用。本研究旨在探讨微小 RNA-302b(miR-302b)对肾纤维化的影响,以及 miR-302b 与 TGF-β 信号通路在单侧输尿管梗阻(UUO)模型中的相互作用。通过检索基因表达综合数据库(GEO),下载微阵列数据集 GSE42716。根据生物信息学分析结果,miR-302b 在 UUO 小鼠肾脏组织和 TGF-β1 处理的 HK-2 细胞中显著下调。Masson 三色染色显示 miR-302b 模拟物减少了 UUO 诱导的肾纤维化。miR-302b 模拟物还通过恢复 E-钙黏蛋白的表达和降低α-平滑肌肌动蛋白(α-SMA)的表达,逆转了胶原 I 和α-SMA 的 mRNA 表达增加以及 E-钙黏蛋白表达降低。此外,miR-302b 的上调还通过恢复 E-钙黏蛋白的表达和降低α-SMA 的表达,抑制了 TGF-β1 诱导的 HK-2 细胞上皮间质转化(EMT)。miR-302b 模拟物抑制了 TGF-βR2 的荧光素酶活性和蛋白表达。然而,miR-302b 抑制剂增加了 TGF-βR2 的荧光素酶活性和蛋白表达。同时,miR-302b 模拟物抑制了体内和体外 TGF-βR2 的 mRNA 表达,并降低了 Smad2 和 Smad3 的磷酸化。此外,TGF-βR2 的过表达恢复了 miR-302b 诱导的胶原 I 和α-SMA 表达降低。综上所述,本研究表明,miR-302b 通过靶向 TGF-βR2 抑制 TGF-β/Smad 信号激活,减轻肾纤维化。我们的研究结果表明,提高肾脏 miR-302b 水平可能是预防肾纤维化的一种新的治疗策略。