网站首页期刊介绍通知公告编 委 会投稿须知电子期刊广告合作联系我们
最新消息:
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
作者:湛世本  田耕  陈建平  王平  王祚才 
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.

参考文献:

[1] 许曼珊, 姜婷, 秦盈盈.骨关节炎发病机制研究进展[J]. 国际骨科学杂志, 2020, 41(4):229-233.
[2] 胡中岭, 王佳洋, 崔逸爽, 等.关节软骨损伤的治疗和研究进展[J]. 中国综合临床, 2019(6):566-571.
[3] 许丽梅, 贾良良, 何晓娟, 等.环状RNA在骨关节炎发生发展中的作用[J]. 中医正骨, 2018, 30(7):39-42.
[4] WANG Y, WU C, ZHANG F, et al. Screening for differentially expressed circular RNAs in the cartilage of osteoarthritis patients for their diagnostic value[J]. Genet Test Mol Biomarkers, 2019, 23(10):706-716.
[5] WANG Y, WU C, YANG Y, et al. Preliminary exploration of hsa_circ_0032131 levels in peripheral blood as a potential diagnostic biomarker of osteoarthritis[J]. Genet Test Mol Biomarkers, 2019, 23(10):717-721.
[6] 李灿锋, 尤田, 沈彬.微小RNA-140与骨关节炎关系的研究进展[J/OL].中华关节外科杂志(电子版), 2020, 14(1):68-72.
[7] YIN C M, SUEN W C, LIN S, et al. Dysregulation of both miR-140-3p and miR-140-5p in synovial fluid correlate with osteoarthritis severity[J]. Bone Joint Res, 2017, 6(11):612-618.
[8] REN T, WEI P, SONG Q, et al. MiR-140-3p ameliorates the progression of osteoarthritis via targeting CXCR4[J]. Biol Pharm Bull, 2020, 43(5):810-816.
[9] Al-MODAWI R N, BRINCHMANN J E, KARLSEN T A.Multi-pathway protective effects of microRNAs on human chondrocytes in an in vitro model of osteoarthritis[J]. Mol Ther Nucleic Acids, 2019, 17:776-790.
[10] 张林, 陆军, 李永刚.骨关节炎中介导软骨细胞代谢失衡的相关信号通路研究进展[J]. 东南大学学报(医学版), 2013, 32(4):465-472.
[11] 柳海平, 周明旺, 李盛华, 等.骨关节炎软骨细胞增殖相关信号通路研究进展[J]. 中国矫形外科杂志, 2018, 26(23):2163-2166.
[12] JENEI-LANZL Z, MEURER A, ZAUCKE F.Interleukin-1β signaling in osteoarthritis-chondrocytes in focus[J]. Cell Signal, 2019, 53:212-223.
[13] 徐传慧, 潘昕, 李超峰, 等.骨关节炎患者血清中软骨代谢标记物与骨关节炎的关系研究[J]. 中国实验诊断学, 2019, 23(2):273-275.
[14] LI B F, ZHANG Y, XIAO J, et al. Hsa_circ_0045714 regulates chondrocyte proliferation, apoptosis and extracellular matrix synthesis by promoting the expression of miR-193b target gene IGF1R[J]. Human Cell, 2017, 30(4):311-318.
[15] ZHOU X, JIANG L, FAN G, et al. Role of the ciRS-7/miR-7 axis in the regulation of proliferation, apoptosis and inflammation of chondrocytes induced by IL-1β[J]. Int Immunopharmacol, 2019, 71:233-240.
[16] NTOUMOU E, TZETIS M, BRAOUDAKI M, et al. Serum microRNA array analysis identifies miR-140-3p, miR-33b-3p and miR-671-3p as potential osteoarthritis biomarkers involved in metabolic processes[J]. Clin Epigenetics, 2017, 9:127.
[17] RASHEED Z, RASHEED N, Al-SHAYA O.Epigallocatechin-3-O-gallate modulates global microRNA expression in interleukin-1β-stimulated human osteoarthritis chondrocytes:potential role of EGCG on negative co-regulation of microRNA-140-3p and ADAMTS5[J]. Eur J Nutr, 2018, 57(3):917-928.
[18] ZHONG S, OUYANG Q, ZHU D, et al. Hsa_circ_0088036 promotes the proliferation and migration of fibroblast-like synoviocytes by sponging miR-140-3p and upregulating SIRT 1 expression in rheumatoid arthritis[J]. Mol Immunol, 2020, 125:131-139.

服务与反馈:
文章下载】【发表评论】【查看评论】【加入收藏
提示:您还未登录,请登录!点此登录
您是第 510041 位访问者


 ©《现代医学》编辑部
联系电话:025-83272481;83272479
电子邮件: xdyx@pub.seu.edu.cn

苏ICP备09058541