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MIR-140-5p affects chondrocyte proliferation, apoptosis, and inflammation by targeting HMGB1 in osteoarthritis.
Inflammation Research ( IF 6.7 ) Pub Date : 2019-11-11 , DOI: 10.1007/s00011-019-01294-0
Yingjie Wang 1 , Songpo Shen 2 , Zeng Li 1 , Weifeng Li 1 , Xisheng Weng 1
Affiliation  

OBJECTIVE This study aimed to test the expression and biological function of miR-140-5p in osteoarthritis (OA), and identify its target gene and explore its mechanism in OA. METHODS Differential genes were screened and analyzed by gene microarray and WGCNA analysis. The normal human chondrocytes C28/I2 were induced by IL-1β to construct the OA cell model. The expression of miR-140-5p and high mobility group box 1 (HMGB1) was quantified by quantitative real-time PCR (qRT-PCR) in OA tissues and IL-1β-induced chondrocytes. Western blotting was performed to evaluate the expression of HMGB1 and PI3K/AKT pathway activation. The concentrations of tumor necrosis factor (TNF)-α, interleukin (IL)-6, MMP-1 and MMP-3 were determined by ELISA. CCK-8 and flow cytometry were conducted to determine the cellular capabilities of proliferation and cell apoptosis. RESULTS Bioinformatics analysis demonstrated that HMGB1 was highly expressed in OA and activated PI3K/AKT pathway. Also, HMGB1 was predicted as a target of miR-140-5p. The levels of miR-140-5p were negatively correlated with HMGB1 in OA tissues and IL-1β-induced chondrocytes. The overexpression of miR-140-5p reduced the expression of HMGB1 protein, p-AKT (Ser473) and p-PI3K in IL-1β-induced chondrocytes. Besides, the expression of p-AKT (Ser473) and p-PI3K was significantly upregulated by employing miR-140-5p inhibitor, but retrieved after treating with LY294002. Furthermore, miR-140-5p inhibited inflammation, matrix metalloprotease expression and apoptosis in IL-1β-induced chondrocytes through regulating HMGB1. CONCLUSION MiR-140-5p was down-regulated while HMGB1 was upregulated in OA. MiR-140-5p could inhibit the PI3K/AKT signaling pathway and suppress the progression of OA through targeting HMGB1.

中文翻译:

MIR-140-5p 通过靶向骨关节炎中的 HMGB1 影响软骨细胞增殖、凋亡和炎症。

目的本研究旨在检测miR-140-5p在骨关节炎(OA)中的表达及生物学功能,鉴定其靶基因并探讨其在OA中的作用机制。方法通过基因芯片和WGCNA分析筛选和分析差异基因。IL-1β诱导正常人软骨细胞C28/I2构建OA细胞模型。通过OA组织和IL-1β诱导的软骨细胞中的定量实时PCR(qRT-PCR)定量miR-140-5p和高迁移率组框1(HMGB1)的表达。进行蛋白质印迹以评估 HMGB1 的表达和 PI3K/AKT 通路激活。ELISA法测定肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6、MMP-1和MMP-3的浓度。进行CCK-8和流式细胞术以确定细胞增殖和细胞凋亡的能力。结果生物信息学分析表明HMGB1在OA中高表达并激活PI3K/AKT通路。此外,HMGB1 被预测为 miR-140-5p 的靶标。miR-140-5p 的水平与 OA 组织和 IL-1β 诱导的软骨细胞中的 HMGB1 呈负相关。miR-140-5p 的过表达降低了 IL-1β 诱导的软骨细胞中 HMGB1 蛋白、p-AKT (Ser473) 和 p-PI3K 的表达。此外,通过使用 miR-140-5p 抑制剂,p-AKT (Ser473) 和 p-PI3K 的表达显着上调,但在 LY294002 处理后恢复。此外,miR-140-5p 通过调节 HMGB1 抑制 IL-1β 诱导的软骨细胞炎症、基质金属蛋白酶表达和凋亡。结论 MiR-140-5p在OA中下调而HMGB1上调。
更新日期:2019-11-01
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