Circ_0114428通过调节miR-185-5p/SMAD3轴促进高糖诱导的肾小球系膜细胞的增殖、纤维化和EMT过程。

IF 3.3 4区 医学 Q3 IMMUNOLOGY
Autoimmunity Pub Date : 2022-11-01 Epub Date: 2022-07-26 DOI:10.1080/08916934.2022.2103797
Bo Li, Guijiang Sun, Haibo Yu, Jia Meng, Fang Wei
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引用次数: 4

摘要

环状RNA (circRNA)已被证实是糖尿病肾病(DN)进展的关键调节因子。然而,circ_0114428在DN进展中的作用尚不清楚。用高糖(HG)处理肾小球系膜细胞(GMCs)以模拟体外DN细胞模型。采用实时荧光定量PCR检测circ_0114428、microRNA (miR)-185-5p、SMAD3 mRNA的表达水平。采用MTT法、EdU染色法和流式细胞术检测细胞增殖能力。western blot法检测细胞增殖标志物、纤维化标志物、上皮-间质转化(epithelial-mesenchymal transition, EMT)标志物和SMAD3蛋白水平。miR-185-5p与circ_0114428或SMAD3之间的相互作用通过双荧光素酶报告基因实验、RIP实验和RNA下拉实验证实。我们的数据显示circ_0114428在hg诱导的gmc中上调。Circ_0114428过表达可加重HG对GMCs增殖、纤维化和EMT过程的促进作用,而其敲低则相反。从机制上看,circ_0114428可以海绵miR-185-5p调控SMAD3。MiR-185-5p抑制剂可逆转circ_0114428敲低对hg诱导的gmc增殖、纤维化和EMT过程的抑制作用。此外,SMAD3过表达消除了miR-185-5p对hg诱导的GMCs增殖、纤维化和EMT过程的抑制作用。综上所述,我们的数据表明circ_0114428可能通过调节miR-185-5p/SMAD3轴促进DN进展。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Circ_0114428 promotes proliferation, fibrosis and EMT process of high glucose-induced glomerular mesangial cells through regulating the miR-185-5p/SMAD3 axis.

Circular RNA (circRNA) has been confirmed to be the key regulators of diabetic nephropathy (DN) progression. However, the role of circ_0114428 in the DN progression remains unclear. Glomerular mesangial cells (GMCs) were treated with high glucose (HG) to mimic DN cell models in vitro. The expression levels of circ_0114428, microRNA (miR)-185-5p, and SMAD3 mRNA were examined by quantitative real-time PCR. Cell proliferation ability was detected by MTT assay, EdU staining and flow cytometry. The protein levels of proliferation marker, fibrosis markers, epithelial-mesenchymal transition (EMT) markers and SMAD3 were measured by western blot assay. The interaction between miR-185-5p and circ_0114428 or SMAD3 was confirmed via dual-luciferase reporter assay, RIP assay and RNA pull-down assay. Our data showed that circ_0114428 was upregulated in HG-induced GMCs. Circ_0114428 overexpression could aggravate the promotion effect of HG on the proliferation, fibrosis and EMT process of GMCs, while its knockdown had an opposite effect. In the terms of mechanisms, circ_0114428 could sponge miR-185-5p to regulate SMAD3. MiR-185-5p inhibitor could reverse the suppressive effect of circ_0114428 knockdown on the proliferation, fibrosis and EMT process in HG-induced GMCs. Also, SMAD3 overexpression abolished the inhibition of miR-185-5p on the proliferation, fibrosis and EMT process in HG-induced GMCs. Taken together, our data suggested that circ_0114428 might promote DN progression by regulating the miR-185-5p/SMAD3 axis.

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来源期刊
Autoimmunity
Autoimmunity 医学-免疫学
CiteScore
5.70
自引率
8.60%
发文量
59
审稿时长
6-12 weeks
期刊介绍: Autoimmunity is an international, peer reviewed journal that publishes articles on cell and molecular immunology, immunogenetics, molecular biology and autoimmunity. Current understanding of immunity and autoimmunity is being furthered by the progress in new molecular sciences that has recently been little short of spectacular. In addition to the basic elements and mechanisms of the immune system, Autoimmunity is interested in the cellular and molecular processes associated with systemic lupus erythematosus, rheumatoid arthritis, Sjogren syndrome, type I diabetes, multiple sclerosis and other systemic and organ-specific autoimmune disorders. The journal reflects the immunology areas where scientific progress is most rapid. It is a valuable tool to basic and translational researchers in cell biology, genetics and molecular biology of immunity and autoimmunity.
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