Upregulation of miR-101a Suppresses Chronic Renal Fibrosis by Regulating KDM3A via Blockade of the YAP-TGF-β-Smad Signaling Pathway
Upregulation of miR-101a Suppresses Chronic Renal Fibrosis by Regulating KDM3A via Blockade of the YAP-TGF-β-Smad Signaling Pathway
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DOI：
10.1016/j.omtn.2020.01.002
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发表时间：
2020-03-06
影响因子：
8.8
通讯作者：
Jiang, Nan
中科院分区：
文献类型：
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作者：
Ding, Hong;Xu, Yanyan;Jiang, Nan
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（miRNAs或miRs）与肾纤维化之间存在关联，但已有报道称miRNAs在慢性肾纤维化的发展中起着至关重要的作用。因此，本研究的目的是探讨miR-101 a在慢性肾纤维化中的可能功能。首先，基于基因芯片的肾纤维化基因表达谱被用来筛选差异表达的基因。建立了单侧输尿管梗阻（UUO）致小鼠慢性肾纤维化的体内模型和马兜铃酸（AA）诱导的体外细胞模型。使用荧光原位杂交（FISH）测定和定量逆转录聚合酶链反应（qRT-PCR）检测miR-101 a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-101 a和KDM 3A之间的相互作用。最后，进行功能获得和丧失实验以阐明miR 101 a对Col 1a 1、纤连蛋白、α-平滑肌肌动蛋白（α-SMA）和YAP-TGF-β（转化生长因子β）-Smad信号通路相关基因表达以及肾纤维化程度的影响。在慢性肾纤维化组织和细胞中，miR-101 a表达较低，而KDM 3A被强烈诱导。此外，miR-101 a可靶向下调KDM 3A的表达，从而导致TGF-1升高，抑制I型胶原（Col 1 a1）、纤连蛋白、α-SMA、YAP 1和TGF-β 2的表达，沿着Smad 2/3磷酸化程度的增加，并延缓肾纤维化程度。此外，过表达雅普/TGF-β 2或抑制TGF-β 1可部分恢复miR-101 a对慢性肾纤维化的抑制作用。总之，miR-101 a可以通过KDM 3A灭活YAP-TGF-β-Smad信号通路来潜在地减缓慢性肾纤维化，突出了miR-101 a作为慢性肾纤维化治疗的治疗靶点的潜力。
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 miR101a on the expression of Col1a1, fibronectin, alpha-smooth muscle actin (alpha-SMA), and YAP-TGF-beta (transforming growth factor beta)-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 (Col1 a1), fibronectin, alpha-SMA, YAP1, and TGF-beta 2 along with the extent of Smad2/3 phosphorylation, as well as delayed renal fibrosis degree. Besides, overexpressed YAP/TGF-beta 2 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-beta-Smad signaling pathway via KDM3A, highlighting the potential of miR-101a as a therapeutic target for chronic renal fibrosis treatment.