miR-203基因启动子区甲基化与肥胖膝关节骨性关节炎患者滑膜炎症水平相关性研究

Study on the correlation between the methylation of miR-203 gene promoter region and the level of synovitis in obese patients with knee osteoarthritis

目的研究滑膜组织miR-203基因启动子区异常甲基化状态与肥胖膝关节骨性关节炎(KOA)关节液促炎因子水平相关性。方法 KOA患者滑膜组织及关节液取自2018年1月~2019年1月于河北省邯郸市中心医院行膝关节人工关节置换的36例患者,体重指数(BMI)≥28.0 kg/m^2的患者纳入肥胖组(23例),18.0 kg/m^2≤BMI≤23.9 kg/m^2的患者纳入正常对照组(13例)。甲基化特异性PCR(MSP)检测滑膜组织miR-203的基因启动子区甲基化状态。通过定量聚合酶链反应(qPCR)检测miR-203在滑膜组织的表达水平;应用qPCR检测miR-203在肥胖组和正常对照组中的表达水平;采用Pearson相关分析探讨miR-203与BMI,关节液中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)的相关性。结果肥胖组滑膜组织中miR-203基因启动子区呈异常低甲基化状态,正常对照组则呈部分低甲基化状态。与正常对照组比较,肥胖组滑膜组织中miR-203的表达水平升高,差异有统计学意义(P<0.05)。肥胖组关节液中IL-1β、IL-6、TNFα的相对表达均高于正常对照组,差异均有统计学意义(均P<0.05)。miR-203基因启动子区甲基化水平与人群BMI和关节液中IL-1β、IL-6、TNF-α均呈负相关(均P<0.05);miR-203表达水平和与人群BMI和关节液中IL-1β、IL-6、TNF-α均呈正相关(均P<0.05)。结论 KOA患者滑膜组织miR-203基因启动子区低甲基化与肥胖KOA患者BMI及关节液促炎因子水平密切相关,miR-203可作为肥胖KOA炎症的重要调控因素及KOA治疗的潜在靶标。

Objective To study the correlation between abnormal methylation of miR-203 gene promoter region in synovial tissues and the level of joint fluid proinflammatory factors in obese knee osteoarthritis(KOA).Methods Synovial tissue and joint fluid of KOA patients were taken from 36 patients who underwent knee arthroplasty in Handan Central Hospital in Hebei Province from January 2018 to January 2019.Patients with body mass index(BMI)≥28.0 kg/m^2were included in the obesity group(23 cases),and patients with 18.0 kg/m^2≤BMI≤23.9 kg/m^2were included in the normal control group(13 cases).Methylation specific PCR(MSP)was used to detect the methylation status of the gene promoter region of miR-203 in synovial tissues.The expression level of miR-203 in synovial tissues was determined by quantitative polymerase chain reaction(qPCR).qPCR was used to detect the expression levels of miR-203 in the obese group and the normal control group.Pearson correlation analysis was used to investigate the correlation between miR-203 and BMI,tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)in joint fluid.Results In the synovial tissues of the obese group,the promoter region of miR-203 gene was abnormally hypomorphic,while in the normal control group,it was partially hypomorphic.Compared with the normal control group,the expression level of miR-203 in synovial tissues of the obesity group was increased,and the difference was statistically significant(P<0.05).The relative expressions of IL-1β,IL-6 and TNF-αin the joint fluid of the obese group were higher than those of the normal control group,with statistically significant differences(all P<0.05).The methylation level in the promoter region of miR-203 gene was negatively correlated with population BMI and the IL-1β,IL-6 and TNF-αin the joint fluid(all P<0.05).The expression level of miR-203 was positively correlated with population BMI and the IL-1β,IL-6 and TNF-αin the joint fluid(all P<0.05).Conclusion Hypophalation of miR-203 gene promoter in synovial tissues of KOA patients is closely related to BMI and levels of joint fluid proinflammatory factors in obese KOA patients,and miR-203 can be used as an important regulatory factor for inflammation of obese KOA and a potential target for treatment of KOA.