Abstract：
BACKGROUND: Although microRNA (miR)-150-5p participates in the progression of renal fibrosis, its mechanism of action remains elusive.
METHODS: A mouse model of unilateral ureteral obstruction was used. The in vitro renal fibrosis model was established by stimulating human kidney 2 (HK-2) cells with transforming growth factor beta 1 (TGF-β1). The expression profiles of miR-150-5p, zinc finger E-box binding homeobox 1 (ZEB1), and other fibrosis- and epithelial-mesenchymal transition (EMT)-linked proteins were determined using Western blot and quantitative reverse transcription polymerase chain reaction. The relationship between miR-150-5p and ZEB1 in HK-2 cells was confirmed by a dual-luciferase reporter assay.
RESULTS: Both in vivo and in vitro renal fibrosis models revealed reduced miR-150-5p expression and elevated ZEB1 level. A significant decrease in E-cadherin levels, as well as increases in alpha smooth muscle actin (α-SMA) and collagen type I (Col-I) levels, was seen in TGF-β1-treated HK-2 cells. The overexpression of miR-150-5p ameliorated TGF-β1-mediated fibrosis and EMT. Notably, miR-150-5p acts by directly targeting ZEB1. A significant reversal of the inhibitory impact of miR-150-5p on TGF-β1-mediated fibrosis and EMT in HK-2 cells was observed upon ZEB1 overexpression.
CONCLUSIONS: MiR-150-5p suppresses TGF-β1-induced fibrosis and EMT by targeting ZEB1 in HK-2 cells, providing helpful insights into the therapeutic intervention of renal fibrosis.
METHODS: A mouse model of unilateral ureteral obstruction was used. The in vitro renal fibrosis model was established by stimulating human kidney 2 (HK-2) cells with transforming growth factor beta 1 (TGF-β1). The expression profiles of miR-150-5p, zinc finger E-box binding homeobox 1 (ZEB1), and other fibrosis- and epithelial-mesenchymal transition (EMT)-linked proteins were determined using Western blot and quantitative reverse transcription polymerase chain reaction. The relationship between miR-150-5p and ZEB1 in HK-2 cells was confirmed by a dual-luciferase reporter assay.
RESULTS: Both in vivo and in vitro renal fibrosis models revealed reduced miR-150-5p expression and elevated ZEB1 level. A significant decrease in E-cadherin levels, as well as increases in alpha smooth muscle actin (α-SMA) and collagen type I (Col-I) levels, was seen in TGF-β1-treated HK-2 cells. The overexpression of miR-150-5p ameliorated TGF-β1-mediated fibrosis and EMT. Notably, miR-150-5p acts by directly targeting ZEB1. A significant reversal of the inhibitory impact of miR-150-5p on TGF-β1-mediated fibrosis and EMT in HK-2 cells was observed upon ZEB1 overexpression.
CONCLUSIONS: MiR-150-5p suppresses TGF-β1-induced fibrosis and EMT by targeting ZEB1 in HK-2 cells, providing helpful insights into the therapeutic intervention of renal fibrosis.
摘要：
背景：尽管microRNA（miR）-150-5p参与肾纤维化的进展，其作用机制仍然难以捉摸。
方法：采用单侧输尿管梗阻小鼠模型。用转化生长因子β1(TGF-β1)刺激人肾2(HK-2)细胞建立体外肾纤维化模型。miR-150-5p的表达谱,锌指E盒绑定homeobox1(ZEB1),以及其他纤维化和上皮间质转化（EMT）连接的蛋白使用Westernblot和定量逆转录聚合酶链反应进行测定。HK-2细胞中miR-150-5p和ZEB1之间的关系通过双荧光素酶报告基因测定来证实。
结果：体内和体外肾纤维化模型均显示miR-150-5p表达降低和ZEB1水平升高。E-cadherin水平显着下降，以及α平滑肌肌动蛋白（α-SMA）和I型胶原蛋白（Col-I）水平的增加，在TGF-β1处理的HK-2细胞中观察到。miR-150-5p的过表达改善了TGF-β1介导的纤维化和EMT。值得注意的是,miR-150-5p通过直接靶向ZEB1起作用。在ZEB1过表达后，观察到miR-150-5p对TGF-β1介导的HK-2细胞纤维化和EMT的抑制作用的显着逆转。
结论：MiR-150-5p通过在HK-2细胞中靶向ZEB1抑制TGF-β1诱导的纤维化和EMT，为肾纤维化的治疗干预提供有益的见解。
方法：采用单侧输尿管梗阻小鼠模型。用转化生长因子β1(TGF-β1)刺激人肾2(HK-2)细胞建立体外肾纤维化模型。miR-150-5p的表达谱,锌指E盒绑定homeobox1(ZEB1),以及其他纤维化和上皮间质转化（EMT）连接的蛋白使用Westernblot和定量逆转录聚合酶链反应进行测定。HK-2细胞中miR-150-5p和ZEB1之间的关系通过双荧光素酶报告基因测定来证实。
结果：体内和体外肾纤维化模型均显示miR-150-5p表达降低和ZEB1水平升高。E-cadherin水平显着下降，以及α平滑肌肌动蛋白（α-SMA）和I型胶原蛋白（Col-I）水平的增加，在TGF-β1处理的HK-2细胞中观察到。miR-150-5p的过表达改善了TGF-β1介导的纤维化和EMT。值得注意的是,miR-150-5p通过直接靶向ZEB1起作用。在ZEB1过表达后，观察到miR-150-5p对TGF-β1介导的HK-2细胞纤维化和EMT的抑制作用的显着逆转。
结论：MiR-150-5p通过在HK-2细胞中靶向ZEB1抑制TGF-β1诱导的纤维化和EMT，为肾纤维化的治疗干预提供有益的见解。
