Wang Xinran, Gu Zhoupeng, Huang Yan, Wang Jingyan, Tang Shiqi, Yang Xinyu, Wang Jianwen
Department of Nephrology, The Third Xiangya Hospital, Central South University, Changsha, China.
The Critical Kidney Disease Research Center of Central South University, Changsha, China.
Eur J Med Res. 2024 Dec 28;29(1):631. doi: 10.1186/s40001-024-02248-x.
The involvement of microRNA-668 (miR-668) in the onset and progression of renal fibrosis remains unclear. To this end, we aimed to explore the relevant mechanism of miR-668 in renal fibrosis.
C57BL/6 J male mice were randomly divided into sham-operated, unilateral ureteral obstruction (UUO), and UUO-fenofibrate groups. Based on transfection and drug intervention, HK-2 cells were divided into blank control, TGF-β1, TGF-β1 + fenofibrate (PPARα agonist), mimics-NC, miR-668, mimics-NC + TGF-β1, miR-668 + TGF-β1, miR-668 + TGF-β1 + fenofibrate, miR-668 + TGF-β1 + GW6471 (PPARα inhibitor), mimics-NC + TGF-β1 + fenofibrate, and mimics-NC + TGF-β1 + GW6471 groups. The pathological changes in the renal tissues were observed by hematoxylin-eosin (HE) and Masson staining. The expression of PPARα, PGC-1α, miR-668, E-cadherin, Collagen III (Col III), and α-SMA in the renal tissues or HK-2 cells was detected by western blot, immunohistochemical analyses or real-time quantitative polymerase chain reaction. The regulatory effect of miR-668 on PPARα was verified by dual-luciferase reporter assay.
The expression of PPARα and PGC-1α decreased in UUO mice and TGF-β1-induced HK-2 cells, which was improved by fenofibrate. Compared to the non-transfected group, in TGF-β1-stimulated HK-2 cells, the expression of E-cadherin, PPARα and PGC-1α increased and the expression of Col III and α-SMA decreased in the miR-668-transfected group. The dual-luciferase reporter assay indicated the regulatory effect of hsa-mir-668-3p on PPARα.
MiR-668 can target PPARα and positively regulate the PPARα/PGC-1α pathway to alleviate renal fibrosis.
微小RNA-668（miR-668）在肾纤维化的发生和发展中的作用尚不清楚。为此，我们旨在探讨miR-668在肾纤维化中的相关机制。
将C57BL/6 J雄性小鼠随机分为假手术组、单侧输尿管梗阻（UUO）组和UUO-非诺贝特组。基于转染和药物干预，将HK-2细胞分为空白对照组、转化生长因子-β1（TGF-β1）组、TGF-β1+非诺贝特（过氧化物酶体增殖物激活受体α激动剂）组、模拟物阴性对照组、miR-668组、模拟物阴性对照组+TGF-β1组、miR-668+TGF-β1组、miR-668+TGF-β1+非诺贝特组、miR-668+TGF-β1+GW6471（过氧化物酶体增殖物激活受体α抑制剂）组、模拟物阴性对照组+TGF-β1+非诺贝特组和模拟物阴性对照组+TGF-β1+GW6471组。通过苏木精-伊红（HE）染色和Masson染色观察肾组织的病理变化。采用蛋白质免疫印迹法、免疫组织化学分析或实时定量聚合酶链反应检测肾组织或HK-2细胞中过氧化物酶体增殖物激活受体α（PPARα）、过氧化物酶体增殖物激活受体γ共激活因子1α（PGC-1α）、miR-668、E-钙黏蛋白、Ⅲ型胶原（ColⅢ）和α-平滑肌肌动蛋白（α-SMA）的表达。通过双荧光素酶报告基因检测验证miR-668对PPARα的调控作用。
在UUO小鼠和TGF-β1诱导的HK-2细胞中，PPARα和PGC-1α的表达降低，非诺贝特可改善这种情况。与未转染组相比，在TGF-β1刺激的HK-2细胞中，miR-668转染组中E-钙黏蛋白、PPARα和PGC-1α的表达增加，ColⅢ和α-SMA的表达降低。双荧光素酶报告基因检测表明hsa-mir-668-3p对PPARα有调控作用。
MiR-668可靶向PPARα并正向调节PPARα/PGC-1α通路以减轻肾纤维化。
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