推拿治疗骨关节炎的表观遗传调控机制研究  

Epigenetic regulatory mechanism of Tuina in treatment of osteoarthritis

作　　者：王欢 丁海涛[1] 李晨浩 杨帆[1] 贾云芳[1] 唐学章[1] WANG Huan;DING Haitao;LI Chenhao;YANG Fan;JIA Yunfang;TANG Xuezhang(Tuina Department,China-Japan Friendship Hospital,Beijing 100029,China)

机构地区：[1]中日友好医院推拿科,北京100029

出　　处：《北京中医药》2025年第3期289-295,共7页Beijing Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金青年项目(81804214);中央高水平医院临床科研业务费资助项目(2022-NHLHCRF-LX-02-0106);北京市自然科学基金面上项目(7242259)。

摘　　要：目的 观察推拿治疗骨关节炎(OA)的表观遗传调控机制及其对滑膜炎症和软骨修复的作用。方法 将30只大鼠采用随机数字表法分为正常组、假手术组、模型组、推拿组、阳性对照组。正常组不进行任何处理;假手术组仅暴露关节囊,但不切断韧带;其余3组均用膝关节前交叉韧带切断术建立OA模型。造模后,正常组、假手术组、模型组不接受任何干预。推拿组在造模后给予推拿治疗,阳性对照组给予关节腔内注射倍他米松复配液,疗程均为4周。末次干预结束1周后,用蛋白质免疫印迹法检测各组滑膜组织中炎症因子[白细胞介素(IL)-10、IL-1β、肿瘤坏死因子α(TNF-α)]、关节软骨中基质蛋白[Ⅱ型胶原蛋白(COL-2)、聚集蛋白聚糖(ACAN)]及表观遗传学关键酶TET1的表达;用甲基化特异性PCR(MSP)分析IL-10启动子区的甲基化水平;用免疫荧光法观察软骨组织中核因子kappa-B(NF-κB)的核转位情况及荧光强度。结果 与正常组、假手术组比较,模型组滑膜组织中IL-10表达低(P<0.01),IL-1β、TNF-α表达高(P<0.01);软骨组织中COL-2、ACAN表达低(P<0.01)。与模型组比较中,推拿组和阳性对照组上述指标均显著改善(P<0.05,P<0.01),其中推拿组IL-10、COL-2、ACAN的表达优于阳性对照组(P<0.05)。与正常组、假手术组比较,模型组滑膜组织中TET1表达低(P<0.01)。与模型组比较,推拿组滑膜组织中TET1表达高(P<0.01),阳性对照组滑膜组织中TET1表达差异无统计学意义(P>0.05)。模型组中IL-10启动子区甲基化条带显著增强,非甲基化条带明显减弱。推拿组中IL-10启动子区非甲基化条带信号有所增强,甲基化条带明显减弱。阳性对照组IL-10启动子区的甲基化和非甲基化条带信号均未见显著变化。模型组中NF-κB的核内信号显著增强,整体荧光强度明显升高。推拿组中NF-κB的核内信号显著减弱,整体荧光强度明显下降。阳性对照组中虽然NF-Objective To observe the epigenetic regulatory mechanism of Tuina in osteoarthritis(OA)and its effects on synovial inflammation and cartilage repair.Methods Thirty rats were randomly assigned to five groups:normal group,sham operation group,model group,Tuina group,and positive control group according to the random number table.The normal group received no treatment.The sham operation group only exposed the joint capsule without cutting the ligaments.The remaining three groups underwent anterior cruciate ligament transection to establish the OA model.After modeling,the normal group,sham operation group,and model group received no further intervention.The Tuina group received manual therapy after modeling,and the positive control group was administered an intra-articular injection of betamethasone compound solution.The treatment lasted for 4 weeks.One week after the final intervention,Western blot(WB)was used to detect the expression of inflammatory factors(IL-10,IL-1β,TNF-α)in synovial tissue,cartilage matrix proteins(COL-2,ACAN)in articular cartilage,and the key epigenetic enzyme TET1.Methylationspecific PCR(MSP)was used to analyze the methylation level of the IL-10 promoter region.Immunofluorescence(IF)was employed to observe NF-κB nuclear translocation and fluorescence intensity in cartilage.Results Compared with the normal and sham operation groups,the model group showed significantly lower IL-10 expression(P<0.01)and higher IL-1βand TNF-αexpression(P<0.01)in synovial tissue,and lower COL-2 and ACAN expression(P<0.01)in cartilage.Compared with the model group,the Tuina group and positive control group showed significant improvements in these indicators(P<0.05,P<0.01),with the Tuina group showing superior recovery of IL-10,COL-2,and ACAN expression compared to the positive control group(P<0.05).The model group exhibited significantly lower TET1 expression in synovial tissue compared with the normal and sham operation groups(P<0.01).The Tuina group showed higher TET1 expression in synovial tissue compared w

关 键 词：推拿 DNA去甲基化 表观遗传 骨关节炎 大鼠 

分 类 号：R244.1[医药卫生—针灸推拿学]