Shenzhen Traditional Chinese Medicine Anorectal Hospital (FuTian), No. 1 Songling Road, Futian District, 518000, Shenzhen, China.
Suzhou Science and Technology City Hospital Affiliated to Nanjing Medical University, 215000, Suzhou, China.
J Mol Histol. 2021 Dec;52(6):1215-1224. doi: 10.1007/s10735-021-10021-1. Epub 2021 Oct 8.
Tight junction proteins play crucial roles in maintaining the integrity of intestinal mucosal barrier. MiRNA-182-5p is capable of targeting claudin-2 which is one of the vital tight junction proteins and the effect and mechanism of miRNA-182-5p was explored here in the DSS-induced colitis model. The pathological conditions were evaluated via hematoxylin and eosin staining. The gene expression level was assessed via PCR. Quantitative immunohistochemistry analysis was performed for the measurement of claudin-2. microRNA.org online tool was used for target gene prediction. Luciferase reporter assay and RNA pull-down assay were performed to detect the target of miRNA-182-5p. The inflammatory and oxidative stress level were measured using corresponding kits. MiRNA-182-5p was highly expressed in colitis model and miRNA-182-5p inhibitor exerted protective effects on colitis induced by DSS in mice. The protective effects includded improvement of pathological changes, increases in anti-inflammation and anti-oxidative genes, and up-regulation of TGF-β1. Claudin-2 mRNA was predicted as the target of miRNA-182-5p, which was validated via luciferase reporter assay and RNA pull-down assay. Claudin-2 overexpression was found in miRNA-182-5p inhibitor group. Consistent with the role of miRNA-182-5p, claudin-2 overexpression also exerted protective effects on DSS-induced colitis in mice. Inhibition of miRNA-182-5p exerted protective effects on colitis via targeting and upregulating claudin-2. The findings in study provide a new therapeutic strategy for colitis treatment and lay the foundation for future study.
紧密连接蛋白在维持肠道黏膜屏障完整性方面发挥着关键作用。miRNA-182-5p 能够靶向紧密连接蛋白之一的 claudin-2，本研究探讨了 miRNA-182-5p 在 DSS 诱导的结肠炎模型中的作用和机制。通过苏木精和伊红染色评估病理状况。通过 PCR 评估基因表达水平。通过定量免疫组化分析测量 claudin-2。使用在线工具 microRNA.org 预测靶基因。通过荧光素酶报告基因检测和 RNA 下拉实验检测 miRNA-182-5p 的靶基因。使用相应试剂盒测量炎症和氧化应激水平。miRNA-182-5p 在结肠炎模型中高表达，miRNA-182-5p 抑制剂对 DSS 诱导的小鼠结肠炎具有保护作用。保护作用包括改善病理变化、增加抗炎和抗氧化基因、上调 TGF-β1。 Claudin-2 mRNA 被预测为 miRNA-182-5p 的靶基因，通过荧光素酶报告基因检测和 RNA 下拉实验进行验证。miRNA-182-5p 抑制剂组中发现 Claudin-2 过表达。与 miRNA-182-5p 的作用一致，Claudin-2 过表达也对 DSS 诱导的小鼠结肠炎具有保护作用。抑制 miRNA-182-5p 通过靶向和上调 Claudin-2 对结肠炎发挥保护作用。本研究结果为结肠炎治疗提供了新的治疗策略，并为未来的研究奠定了基础。
