Department of Radiology, The First People's Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology, No.157 Jinbi Road, Kunming, 650032, Yunnan, China.
J Thromb Thrombolysis. 2021 Aug;52(2):440-453. doi: 10.1007/s11239-020-02369-0. Epub 2021 Jan 2.
Long non-coding RNA X-inactive specific transcript (LncRNA XIST) is involved in several diseases. However, the molecular mechanism of XIST and its relation with miR-133a-3p in contrast-induced nephropathy (CIN) remained vague. Sprague-Dawley (SD) rats were assigned to Control, Sham, and CIN groups at random (n = 15 for each group). Histological examination on the kidney tissues was performed using hematoxylin and eosin (HE) and periodic acid-Schiff (PAS) staining. Mean serum creatinine (SCr) and blood urea nitrogen (BUN) contents was measured by colorimetric microplate method. Levels of inflammatory cytokines were detected by enzyme-linked immunosorbent assay (ELISA). The cells viability and apoptosis were respectively detected by MTT assay and flow cytometry. Target gene and potential binding sites between XIST, miR-133a-3p and NLR Family Pyrin Domain Containing 3 (NLRP3) were predicted using online databases and confirmed by dual-luciferase reporter assay. Relative mRNA and protein expressions of XIST, miR-133a-3p, NLRP3, apoptosis-associated speck-like protein (ASC) and Cleaved caspase-1 were measured with quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot as needed. In the rat CIN model, Ioversol induced kidney morphology changes, with increase on SCr and BUN contents, elevated levels of inflammatory cytokines and upregulated expressions of XIST, NLRP3, ASC and Cleaved caspase-1. Silencing XIST reversed the effects of Ioversol on cells. MiR-133a-3p could bind with XIST and target NLRP3, and downregulating miR-133a-3p reversed the effect of silencing XIST on Ioversol-treated cells. Moreover, downregulating XIST attenuated CIN injury via regulating miR-133a-3p/NLRP3 axis.
长链非编码 RNA X 染色体失活特异性转录物（LncRNA XIST）参与多种疾病。然而，XIST 的分子机制及其与对比诱导肾病（CIN）中 miR-133a-3p 的关系尚不清楚。SD 大鼠随机分为对照组、假手术组和 CIN 组（每组 15 只）。通过苏木精和伊红（HE）和过碘酸希夫（PAS）染色对肾组织进行组织学检查。比色微板法测定血清肌酐（SCr）和血尿素氮（BUN）含量。酶联免疫吸附试验（ELISA）检测炎症细胞因子水平。MTT 法和流式细胞术分别检测细胞活力和细胞凋亡。利用在线数据库预测 XIST、miR-133a-3p 和富含 N 端亮氨酸重复域的 NLR 家族 Pyrin 结构域 3（NLRP3）之间的靶基因和潜在结合位点，并通过双荧光素酶报告基因检测验证。实时定量聚合酶链反应（qRT-PCR）和 Western blot 法检测 XIST、miR-133a-3p、NLRP3、凋亡相关斑点样蛋白（ASC）和Cleaved caspase-1 的相对 mRNA 和蛋白表达。在大鼠 CIN 模型中，碘海醇诱导肾脏形态改变，SCr 和 BUN 含量增加，炎症细胞因子水平升高，XIST、NLRP3、ASC 和 Cleaved caspase-1 表达上调。沉默 XIST 逆转了碘海醇对细胞的作用。miR-133a-3p 可与 XIST 结合并靶向 NLRP3，下调 miR-133a-3p 可逆转沉默 XIST 对碘海醇处理细胞的作用。此外，下调 XIST 通过调节 miR-133a-3p/NLRP3 轴减轻 CIN 损伤。
