医学分子生物学杂志2024，Vol.21Issue(2)：115-123,9.DOI:10.3870/j.issn.1672-8009.2024.02.004
lncHAGLR通过抑制miR-26a激活NF-κB促进骨关节炎大鼠软骨细胞的炎症反应和细胞凋亡
LncHAGLR Promotes Inflammatory Response and Apoptosis of Chondrocytes in Osteoarthritis Rats by Targeting miR-26a and Acti-vation of NF-κB
摘要
Abstract
Objective To investigate the effect and potential mechanism of long non-coding RNA HAGLR on the inflammatory response and apoptosis of chondrocytes in rats with osteoarthri-tis.Methods StarBase and luciferase gene reporter assay were used to predict and verify the inter-action between lncHAGLR and miR-26a.To investigate the regulatory role of lncHAGLR on miR-26a expression,C518 cells were divided into 4 groups:si-lncHAGLR group,si-NC group,miR-26a-inhibitor group,and inhibitor-NC group.Furthermore,to study whether lncHAGLR regulates in-flammation and apoptosis in C518 cells by modulating miR-26a,C518 cells were divided into 5 groups:Control group,IL-1β group,IL-1β + si-NC group,IL-1β + si-lncHAGLR group,and IL-1β + si-lncHAGLR + miR-26a-inhibitor group.Real-time quantitative PCR(qRT-PCR)was used to analyze the expression levels of lncHAGLR and miR-26a.The proliferation,cytotoxicity and apoptosis of C518 cells were detected by MTT,lactate dehydrogenase(LDH)Kit and flow cytome-try(FCM).Western blotting assay was used to detect the protein expression levels of cleaved-CASPASE3,NF-κB P65,and phosphorylated NF-κB P65(P-NF-κB P65).The levels of released inflammatory factors(TNF-α and IL-6)were detected by ELISA.Results lncHAGLR directly tar-geted miR-26a.The expression level of lncHAGLR in the IL-1β group was significantly higher than that in the Control group,and the expression level of miR-26a was significantly lower than that in the Control group(all P<0.05).In addition,cells in the IL-1β +si-lncHAGLR group had reduced IL-1β-induced chondrocyte inflammation,inhibited apoptosis,increased cell viabilities,decreased release of LDH,inhibited expression of cleaved-CASPASE3,and decreased secretions of TNF-α and IL-6(all P<0.05).Moreover,the expression level of P-NF-κB P65 was decreased(P<0.05).Adding of miR-26a-inhibitor(IL-1β + si-lncHAGLR + miR-26a-inhibitor group)reversed the performances of cells in the IL-1β +si-lncHAGLR group(all P<0.05).Conclusion lncHA-GLR promotes the inflammatory response and apoptosis of chondrocytes in osteoarthritis rats by in-hibiting the activation of NF-κB through targeting miR-26a,suggesting that lncHAGLR may be a valuable therapeutic target for osteoarthritis treatment.关键词
lncHAGLR/骨关节炎/C518细胞/细胞凋亡/miR-26a/NF-κB P65Key words
lncHAGLR/osteoarthritis/C518 cells/cell apoptosis/miR-26a/NF-κB P65分类
医药卫生引用本文复制引用
孟晓源,乌尔坎·叶尔肯,王志刚,马乐..lncHAGLR通过抑制miR-26a激活NF-κB促进骨关节炎大鼠软骨细胞的炎症反应和细胞凋亡[J].医学分子生物学杂志,2024,21(2):115-123,9.基金项目
新疆维吾尔自治区自然科学基金(No.2022D01C507) This work was supported by a grant from the Natural Science Foundation of Xinjiang Uygur Autonomous Region(No.2022D01C507) （No.2022D01C507）
