Renal fibrosis denotes a common complication of diabetic nephropathy and is a predominant cause of end-stage renal disease. Despite the association between microRNAs (miRNAs or miRs) and renal fibrosis, miRNAs have been reported to play a vital role in the development of chronic renal fibrosis. Therefore, the aim of the present study was to investigate the possible function of miR-101a in chronic renal fibrosis. Initially, microarray-based gene expression profiling of renal fibrosis was employed to screen the differentially expressed genes. An mouse model of chronic renal fibrosis induced by a unilateral ureteral obstruction (UUO) and an cell model induced by aristolochic acid (AA) were constructed. miR-101a expression was examined using a fluorescence hybridization (FISH) assay and quantitative reverse transcription polymerase chain reaction (qRT-PCR). Then, the interaction between miR-101a and KDM3A was identified using an online website combined with a dual-luciferase reporter assay. Finally, gain- and loss-of-function experiments were conducted to elucidate the effect of miR-101a on the expression of Col1a1, fibronectin, α-smooth muscle actin (α-SMA), and YAP-TGF-β (transforming growth factor β)-Smad signaling pathway-related genes, as well as the degree of renal fibrosis. miR-101a was poorly expressed while KDM3A was robustly induced in chronic renal fibrosis tissues and cells. In addition, miR-101a could target and downregulate KDM3A expression, which led to elevated TGIF1, inhibited expression of Collagen I (Col1a1), fibronectin, α-SMA, YAP1, and TGF-β2 along with the extent of Smad2/3 phosphorylation, as well as delayed renal fibrosis degree. Besides, overexpressed YAP/TGF-β2 or inhibited TGIF1 partially restored the inhibitory effect of miR-101a on chronic renal fibrosis. Taken together, miR-101a could potentially slow down chronic renal fibrosis by the inactivation of the YAP-TGF-β-Smad signaling pathway via KDM3A, highlighting the potential of miR-101a as a therapeutic target for chronic renal fibrosis treatment.

中文翻译：

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miR-101a 上调通过阻断 YAP-TGF-β-Smad 信号通路调节 KDM3A 抑制慢性肾纤维化


肾纤维化是糖尿病肾病的常见并发症，也是终末期肾病的主要原因。尽管 microRNA（miRNA 或 miR）与肾纤维化之间存在关联，但据报道 miRNA 在慢性肾纤维化的发展中发挥着至关重要的作用。因此，本研究的目的是探讨miR-101a在慢性肾纤维化中的可能功能。最初，采用基于微阵列的肾纤维化基因表达谱来筛选差异表达基因。构建单侧输尿管梗阻（UUO）诱导的慢性肾纤维化小鼠模型和马兜铃酸（AA）诱导的细胞模型。使用荧光杂交 (FISH) 测定和定量逆转录聚合酶链反应 (qRT-PCR) 检查 miR-101a 表达。然后，使用在线网站结合双荧光素酶报告基因测定来鉴定 miR-101a 和 KDM3A 之间的相互作用。最后，进行功能获得和丧失实验以阐明 miR-101a 对 Col1a1、纤连蛋白、α-平滑肌肌动蛋白 (α-SMA) 和 YAP-TGF-β（转化生长因子）表达的影响β)-Smad信号通路相关基因，以及肾纤维化程度。在慢性肾纤维化组织和细胞中，miR-101a 表达较差，而 KDM3A 表达强烈。此外，miR-101a可以靶向并下调KDM3A的表达，从而导致TGIF1升高，抑制I型胶原蛋白（Col1a1）、纤连蛋白、α-SMA、YAP1和TGF-β2的表达以及Smad2/3磷酸化的程度，以及迟发性肾纤维化程度。 此外，过表达YAP/TGF-β2或抑制TGIF1可部分恢复miR-101a对慢性肾纤维化的抑制作用。综上所述，miR-101a 可能通过 KDM3A 灭活 YAP-TGF-β-Smad 信号通路来减缓慢性肾纤维化，这凸显了 miR-101a 作为慢性肾纤维化治疗靶点的潜力。