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 in vivo mouse model of chronic renal fibrosis induced by a unilateral ureteral obstruction (UUO) and an in vitro cell model induced by aristolochic acid (AA) were constructed. miR-101a expression was examined using a fluorescence in situ 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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通过阻断YAP-TGF-β-Smad信号通路来调节KDM3A，miR-101a的上调抑制了慢性肾纤维化。

肾纤维化是糖尿病肾病的常见并发症，并且是终末期肾脏疾病的主要原因。尽管microRNA（miRNA或miRs）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起着至关重要的作用。因此，本研究的目的是研究miR-101a在慢性肾纤维化中的可能功能。最初，基于微阵列的肾纤维化基因表达谱被用来筛选差异表达的基因。构建了由单侧输尿管梗阻（UUO）诱导的慢性肾纤维化的体内小鼠模型和由马兜铃酸（AA）诱导的体外细胞模型。使用荧光原位杂交（FISH）分析和定量逆转录聚合酶链反应（qRT-PCR）检测miR-101a的表达。然后，使用在线网站结合双荧光素酶报告基因测定法鉴定了miR-101a和KDM3A之间的相互作用。最后，进行功能获得和丧失功能实验，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的表达较弱，而KDM3A则被强烈诱导。此外，miR-101a可能靶向并下调KDM3A表达，从而导致TGIF1升高，抑制胶原I（Col1a1）的表达，纤连蛋白，α-SMA，YAP1和TGF-β2以及Smad2 / 3磷酸化程度以及延迟的肾纤维化程度。此外，过表达的YAP /TGF-β2或抑制的TGIF1部分地恢复了miR-101a对慢性肾纤维化的抑制作用。两者合计，miR-101a可能通过经由KDM3A灭活YAP-TGF-β-Smad信号传导途径而减缓慢性肾纤维化，突出了miR-101a作为慢性肾纤维化治疗靶点的潜力。