Epigenetic regulatory mechanism of <italic style="font-style: italic">Tuina</italic> in treatment of osteoarthritis

WANG Huan; DING Haitao; LI Chenhao; YANG Fan; JIA Yunfang; TANG Xuezhang

doi:10.16025/j.1674-1307.2025.03.006

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Epigenetic regulatory mechanism of Tuina in treatment of osteoarthritis

更新时间：2025-04-29

- Epigenetic regulatory mechanism of Tuina in treatment of osteoarthritis
- Beijing Journal of Traditional Chinese Medicine Vol. 44, Issue 3, Pages: 289-295(2025)
- 作者机构：
  
  中日友好医院推拿科，北京 100029
- 作者简介：
- 基金信息：
- DOI：10.16025/j.1674-1307.2025.03.006
  CLC：
- Received：18 December 2024，
  
  Published：25 March 2025
- 稿件说明：
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王欢，丁海涛，李晨浩，等.推拿治疗骨关节炎的表观遗传调控机制研究［J］.北京中医药，2025，44（3）：289-295.

WANG Huan,DING Haitao,LI Chenhao,et al.Epigenetic regulatory mechanism of Tuina in treatment of osteoarthritis[J]. Beijing Journal of Traditional Chinese Medicine,2025,44(03):289-295.
王欢，丁海涛，李晨浩，等.推拿治疗骨关节炎的表观遗传调控机制研究［J］.北京中医药，2025，44（3）：289-295. DOI： 10.16025/j.1674-1307.2025.03.006.

WANG Huan,DING Haitao,LI Chenhao,et al.Epigenetic regulatory mechanism of Tuina in treatment of osteoarthritis[J]. Beijing Journal of Traditional Chinese Medicine,2025,44(03):289-295. DOI： 10.16025/j.1674-1307.2025.03.006.

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（

0.01），IL-1β、TNF-α表达高（

0.01）；软骨组织中COL-2、ACAN表达低（

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（

0.05，

0.01），其中推拿组IL-10、COL-2、ACAN的表达优于阳性对照组（

0.05）。与正常组、假手术组比较，模型组滑膜组织中TET1表达低（

0.01）。与模型组比较，推拿组滑膜组织中TET1表达高（

0.01），阳性对照组滑膜组织中TET1表达差异无统计学意义（

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-κB的核内信号有所减少，但整体荧光强度变化有限。

结论

推拿通过TET1介导的DNA去甲基化上调IL-10的表达，抑制NF-κB活性，缓解骨关节炎滑膜炎症，并促进软骨修复。

Abstract

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. Methylation-specific 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 （

0.01） and higher IL-1β and TNF-α expression （

0.01） in synovial tissue， and lower COL-2 and ACAN expression （

0.01） in cartilage. Compared with the model group， the

Tuina

group and positive control group showed significant improvements in these indicators （

0.05，

0.01）， with the

Tuina

group showing superior recovery of IL-10， COL-2， and ACAN expression compared to the positive control group （

0.05）. The model group exhibited significantly lower TET1 expression in synovial tissue compared with the normal and sham operation groups （

0.01）. The

Tuina

group showed higher TET1 expression in synovial tissue compared with the model group （

0.01）， while the positive control group showed no significant change （

0.05）. In the model group， the methylated bands of the IL-10 promoter region were significantly enhanced， while the unmethylated bands were markedly reduced. In the

Tuina

group， the unmethylated bands of the IL-10 promoter region showed enhanced signals， and the methylated bands were significantly reduced. No significant changes were observed in the methylated and unmethylated bands of the IL-10 promoter region in the positive control group. The model group showed significantly enhanced NF-κB nuclear signals and increased overall fluorescence intensity. The

Tuina

group significantly reduced NF-κB nuclear signals and fluorescence intensity. Although the nuclear signal of NF-κB in the positive control group decreased somewhat， the overall fluorescence intensity change was limited.

Conclusion

Tuina

upregulates IL-10 expression through TET1-mediated DNA demethylation， suppresses NF-κB activity， alleviates synovial inflammation in osteoarthritis， and promotes cartilage repair.

关键词

Keywords

references

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⁰

Epigenetic regulatory mechanism of Tuina in treatment of osteoarthritis

DOI：10.16025/j.1674-1307.2025.03.006

摘要

Abstract

关键词

Keywords

references