circHIPK3 Exacerbates Folic Acid-Induced Renal Tubulointerstitial Fibrosis by Sponging miR-30a.
circHIPK3 Exacerbates Folic Acid-Induced Renal Tubulointerstitial Fibrosis by Sponging miR-30a.
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circHIPK3 通过海绵 miR-30a 加剧叶酸诱导的肾小管间质纤维化
DOI：
10.3389/fphys.2021.715567
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发表时间：
2021
影响因子：
4
通讯作者：
Zhou H
中科院分区：
文献类型：
--
作者：
Wu Y;Luan J;Jiao C;Zhang S;Ma C;Zhang Y;Fu J;Lai EY;Kopp JB;Pi J;Zhou H
肾小管间质纤维化是进展性慢性肾脏病（CKD）的常见病理特征，目前的治疗效果有限。据报道，环状RNA circHIPK3参与多种人类疾病的发病机制。然而，circHIPK3在肾纤维化中的作用尚未研究。在这项研究中，我们的目的是确定circHIPK3是否以及如何参与肾纤维化的发病机制。小鼠接受叶酸（250mg/kg）的腹膜注射。值得注意的是，30天后，高碘酸-希夫（PAS）和Masson染色显示肾纤维化，编码纤连蛋白（FN）和胶原1（COL1）的促纤维化基因的mRNA和蛋白质增加。通过定量PCR（qPCR）和荧光原位杂交（FISH）评估，肾circHIPK3上调，而miR-30a下调。通过qPCR分析、免疫印迹和免疫荧光检测，转化生长因子β-1（TGF-β 1）的表达增加。肾circHIPK3与miR-30 a呈负相关，肾miR-30 a与TGF-β 1呈负相关。Target Scan和miRanda算法预测了circHIPK3和miR-30a之间的三个完美结合位点。FISH和免疫荧光染色显示，circHIPK3、miR-30a和TGF-β 1共定位于人肾小管上皮细胞（HK-2细胞）的胞浆中。我们将circHIPK3和一种乱序RNA转染到HK-2细胞中; miR-30a下调，促纤维化基因如TGF-β 1、FN和COL1上调，并通过qPCR、免疫印迹和免疫荧光染色进行评估。第三，在暴露于TGF-β 1的HK-2细胞中还观察到circHIPK3上调、miR-30a下调以及促纤维化FN和COL1的过量产生。最后，慢性肾小管间质性肾炎患者的肾活检显示出与实验模型中观察到的相似的circHIPK3、miR-30 a和促纤维化蛋白（如TGF-β 1、FN和COL1）表达模式。在circHIPK3、miR-30a和TGF-β 1之间观察到前馈循环。我们的研究结果表明，circHIPK3可能有助于通过海绵状miR-30a进行性肾纤维化。circHIPK3可能是减缓CKD进展的新治疗靶点。
Renal tubulointerstitial fibrosis is a common pathological feature of progressive chronic kidney disease (CKD), and current treatment has limited efficacy. The circular RNA circHIPK3 is reported to participate in the pathogenesis of various human diseases. However, the role of circHIPK3 in renal fibrosis has not been examined. In this study, we aimed to determine whether and how circHIPK3 might participate in the pathogenesis of renal fibrosis. Mice received a peritoneal injection of folic acid (250 mg/kg). Of note, 30 days later, renal fibrosis was present on periodic acid–Schiff (PAS) and Masson staining, and mRNA and protein of profibrotic genes encoding fibronectin (FN) and collagen 1 (COL1) were increased. Renal circHIPK3 was upregulated, while miR-30a was downregulated, assessed by quantitative PCR (qPCR) and fluorescence in situ hybridization (FISH). The expression of transforming growth factor beta-1 (TGF-β1) was increased by qPCR analysis, immunoblotting, and immunofluorescence. Renal circHIPK3 negatively correlated with miR-30a, and kidney miR-30a negatively correlated with TGF-β1. Target Scan and miRanda algorithms predicted three perfect binding sites between circHIPK3 and miR-30a. We found that circHIPK3, miR-30a, and TGF-β1 colocalized in the cytoplasm of human tubular epithelial cells (HK-2 cells) on FISH and immunofluorescence staining. We transfected circHIPK3 and a scrambled RNA into HK-2 cells; miR-30a was downregulated, and the profibrotic genes such as TGF-β1, FN, and COL1 were upregulated and assessed by qPCR, immunoblotting, and immunofluorescence staining. Third, the upregulation of circHIPK3, downregulation of miR-30a, and overproduction of profibrotic FN and COL1 were also observed in HK-2 cells exposed to TGF-β1. Finally, renal biopsies from patients with chronic tubulointerstitial nephritis manifested similar expression patterns of circHIPK3, miR-30a, and profibrotic proteins, such as TGF-β1, FN, and COL1 as observed in the experimental model. A feed-forward cycle was observed among circHIPK3, miR-30a, and TGF-β1. Our results suggest that circHIPK3 may contribute to progressive renal fibrosis by sponging miR-30a. circHIPK3 may be a novel therapeutic target for slowing CKD progression.
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