Circ-0032131通过靶向miR-140-3p调控骨关节炎软骨细胞增殖及炎症因子分泌的机制研究-现代医学

Circ-0032131通过靶向miR-140-3p调控骨关节炎软骨细胞增殖及炎症因子分泌的机制研究

单位：琼海市人民医院骨科, 海南琼海 571499

关键词：Circ-0032131 miR-140-3p 骨关节炎软骨细胞增殖炎症因子

分类号：R684.3

出版年·卷·期（页码）：2021·49·第八期（904-910）

摘要：

目的：探讨Circ-0032131对骨关节炎软骨细胞增殖及炎症因子分泌的影响及分子机制。方法：选取本院17例骨关节炎患者的软骨组织以及正常组织为研究对象；将软骨细胞CHON-001分为control组、IL-1β组、IL-1β+si-Circ-0032131组、IL-1β+si-NC组、IL-1β+miR-140-3p mimics组、IL-1β+miR-mimics-NC组、IL-1β+si-NC+miR-inhibitor-NC组、IL-1β+si-Circ-0032131+miR-inhibitor-NC组、IL-1β+si-Circ-0032131+miR-140-3p inhibitor组。实时荧光定量PCR （RT-qPCR）检测Circ-0032131和miR-140-3p的表达水平；Western blot检测蛋白表达；MTT检测细胞增殖；平板实验检测细胞克隆形成数；ELISA检测TNF-α、IL-6水平；双荧光素酶报告实验验证Circ-0032131和miR-140-3p的靶向关系。结果：与正常组织相比，骨关节炎患者软骨组织中Circ-0032131表达水平升高，miR-140-3p表达水平降低（P<0.05）。IL-1β诱导的软骨细胞中MMP1、MMP13表达水平升高，Collagen Ⅱ表达水平降低，细胞OD值降低，细胞克隆形成数降低，CyclinD1表达水平降低，P21表达水平升高，TNF-α、IL-6水平升高（P<0.05）。沉默Circ-0032131或过表达miR-140-3p后，IL-1β诱导的软骨细胞中细胞OD值升高，细胞克隆形成数升高，CyclinD1表达水平升高，P21表达水平降低，TNF-α、IL-6水平降低（P<0.05）。Circ-0032131靶向调控miR-140-3p表达；干扰miR-140-3p能逆转沉默Circ-0032131对骨关节炎软骨细胞增殖及炎症因子分泌的影响。结论：Circ-0032131可能通过靶向miR-140-3p调控IL-1β诱导的软骨细胞增殖及炎症因子分泌。

Objective: To explore the effect of Circ-0032131 on the proliferation and secretion of inflammatory factors in osteoarthritis chondrocytes and its molecular mechanism. Methods: The cartilage tissue of 17 patients with osteoarthritis and normal tissues in our hospital were selected; chondrocytes CHON-001 were divided into control group, IL-1β group, IL-1β+si-Circ-0032131 group, IL-1β+si-NC group, IL-1β+miR-140-3p mimics group, IL-1β+miR-mimics-NC group, IL-1β+si-NC+miR-inhibitor-NC group, IL-1β+si-Circ-0032131+miR-inhibitor-NC group, IL-1β+si-Circ-0032131+miR-140-3p inhibitor group. Real-time fluorescent quantitative PCR(RT-qPCR) was used to detect the expression levels of Circ-0032131 and miR-140-3p; Western blot to detect protein expression; MTT to detect cell proliferation; plate experiment to detect cell clone formation; ELISA to detect TNF-α, IL-6 levels; dual luciferase reporter experiment verified the targeting relationship between Circ-0032131 and miR-140-3p. Results: Compared with normal tissues, the expression level of Circ-0032131 was increased and the expression level of miR-140-3p was decreased in cartilage tissue of patients with osteoarthritis(P<0.05). In chondrocytes induced by IL-1β, the expression levels of MMP1 and MMP13 were increased, the expression level of Collagen II was decreased, the OD value was decreased, the number of cell clone formation was decreased, the expression level of CyclinD1 was decreased, the expression level of P21 was increased, and the levels of TNF-α, IL-6 were increased(P<0.05). After Circ-0032131 silenced or miR-140-3p overexpressed, the OD value was increased, the number of cell clone formation was increased, the expression of CyclinD1 was increased, the expression of P21 was decreased, TNF-α, IL-6 levels were decreased in IL-1β-induced chondrocytes(P<0.05). Circ-0032131 targets miR-140-3p expression; interference with miR-140-3p could reverse the effect of silencing Circ-0032131 on the proliferation and secretion of inflammatory factors in osteoarthritis chondrocytes. Conclusions: Circ-0032131 may regulate IL-1β-induced chondrocyte proliferation and secretion of inflammatory factors by targeting miR-140-3p.

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