浏览全部资源
扫码关注微信
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- 北京中医药 2025年44卷第3期 页码：289-295
- 作者机构：
中日友好医院推拿科，北京 100029
- 作者简介：
王欢，女，38岁，博士，主治医师。研究方向：中医药防治退行性骨关节病。
唐学章，E-mail：tangxuezhang@126.com
- 基金信息：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)
DOI：10.16025/j.1674-1307.2025.03.006
中图分类号：收稿日期：2024-12-18，
纸质出版日期：2025-03-25
- 稿件说明：
移动端阅览
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
2
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
2
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
2
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-κB的核内信号有所减少，但整体荧光强度变化有限。
结论
2
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
2
To observe the epigenetic regulatory mechanism of
Tuina
in osteoarthritis （OA） and its effects on synovial inflammation and cartilage repair.
Methods
2
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
2
Compared with the normal and sham operation groups， the model group showed significantly lower IL-10 expression （
P
<
0.01） and higher IL-1β and TNF-α expression （
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 with the model group （
P
<
0.01）， while the positive control group showed no significant change （
P
>
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
2
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
WANG H , ZHANG Y , ZHANG CF , et al . Exosomes derived from miR-146a-overexpressing fibroblast-like synoviocytes in cartilage degradation and macrophage M1 polarization: a novel protective agent for osteoarthritis [J ] . Front Immunol , 2024 （ 15 ）: 1361606 .
DE ROOVER A , ESCIBANO-NUNEZ A , MONTEAGUDO S , et al . Fundamentals of osteoarthritis: Inflammatory mediators in osteoarthritis [J ] . Osteoarthritis Cartilage , 2023 , 31 ( 10 ): 1303 - 1311 .
PERRUCCIO AV , YOUNG JJ , WILFONG JM , et al . Osteoarthritis year in review 2023:Epidemiology & therapy [J ] . Osteoarthritis Cartilage , 2024 , 32 ( 2 ): 159 - 165 .
GENG RZ , LI JL , YU C , et al . Knee osteoarthritis: Current status and research progress in treatment (Review) [J ] . Exp Ther Med , 2023 , 26 ( 4 ): 481 .
KAN TY , YU ZF , YAN MN . Epigenetic regulation of chondrocytes affected by mitochondria through mechanotransduction in osteoarthritis [J ] . Med Novel Technol Devices , 2023 ( 2 ): 36 - 43 .
YAN S , LU T , YANG H , et al . Decreased histone H3K9 dimethylation in synergy with DNA demethylation of Spi-1 binding site contributes to ADAMTS-5 expression in articular cartilage of osteoarthritis mice [J ] . J Cell Physiol , 2024 , 239 ( 12 ): e31444 .
王欢 , 唐学章 . “舒筋活络”推拿法对骨关节炎固有免疫应答的调控作用 [J ] . 中华中医药杂志 , 2024 , 39 ( 9 ): 4644 - 4648 .
JI LX , JORDAN WT , SHI XL , et al . TET-mediated epimutagenesis of the Arabidopsis thaliana methylome [J ] . Nat Commun , 2018 , 9 ( 1 ): 895 .
WANG H , SHU J , ZHANG CF , et al . Extracellular vesicle-mediated miR-150-3p delivery in joint homeostasis: a potential treatment for osteoarthritis [J ] . Cells , 2022 , 11 ( 17 ): 2766 .
YAO Q , WU XH , TAO C , et al . Osteoarthritis: pathogenic signaling pathways and therapeutic targets [J ] . Signal Transduct Target Ther , 2023 , 8 ( 1 ): 56 .
王欢 , 丁海涛 , 舒峻 , 等 . 舒筋活络方对Toll样受体介导的滑膜炎症调控作用 [J ] . 中华中医药杂志 , 2022 , 37 ( 1 ): 469 - 472 .
CHEN XR , ZHU XB , DONG J , et al . Reversal of epigenetic peroxisome proliferator-activated receptor-γ suppression by diacerein alleviates oxidative stress and osteoarthritis in mice [J ] . Antioxid Redox Signal , 2022 , 37 ( 1-3 ): 40 - 53 .
FENG Y , CHEN JJ , XIE NB , et al . Direct decarboxylation of ten-eleven translocation-produced 5-carboxylcytosine in mammalian genomes forms a new mechanism for active DNA demethylation [J ] . Chem Sci , 2021 , 12 ( 34 ): 11322 - 11329 .
JOSHI K , LIU S , BRESLIN SJP , et al . Mechanisms that regulate the activities of TET proteins [J ] . Cell Mol Life Sci , 2022 , 79 ( 7 ): 363 .
KRIUKIENė E , TOMKUVIENė M , KLIMAšAUSKAS S . 5-Hydroxymethylcytosine: the many faces of the sixth base of mammalian DNA [J ] . Chem Soc Rev , 2024 , 53 ( 5 ): 2264 - 2283 .
王欢 . 中西医结合治疗膝骨关节炎(膝痹)专家共识 [J ] . 世界中医药 , 2023 , 18 ( 17 ): 2407 - 2412 .
王欢 , 唐学章 , 舒骏 , 等 . 基于软骨细胞分离培养的骨痹体外细胞模型的建立 [J ] . 中国中医基础医学杂志 , 2023 , 29 ( 3 ): 401 - 405 .
WANG H , WANG QG , YANG MJ , et al . Histomorphology and innate immunity during the progression of osteoarthritis: Does synovitis affect cartilage degradation [J ] . J Cell Physiol , 2018 , 233 ( 2 ): 1342 - 1358 .
姜一陵 , 季玉婷 , 郑秦 , 等 . 基于TLR/NF-κB信号通路的异功散治疗慢性病贫血的机制研究 [J ] . 北京中医药 , 2024 , 43 ( 10 ): 1100 - 1105 .
MAGIEROWSKA K , WOJCIK-GRZYBEK D , KORBUT E , et al . The mitochondria-targeted sulfide delivery molecule attenuates drugs-induced gastropathy. Involvement of heme oxygenase pathway [J ] . Redox Biol , 2023 （ 66 ）: 102847 .
MAGIEROWSKA K , KORBUT E , WOJCIK-GRZYBEK D , et al . Mitochondria-targeted hydrogen sulfide donors versus acute oxidative gastric mucosal injury [J ] . J Control Release , 2022 （ 348 ）: 321 - 334 .
李金牛 , 霍素坤 , 任振家 , 等 . 针刺结合艾灸、中药热敷治疗膝骨关节炎寒湿痹阻证的临床疗效观察 [J ] . 北京中医药 , 2024 , 43 ( 6 ): 622 - 626 .
王欢 , 王庆甫 , 杨黎黎 , 等 . 从“经筋”理论论治膝骨关节炎滑膜炎症的思路探讨 [J ] . 长春中医药大学学报 , 2017 , 33 ( 1 ): 10 - 12 .
0
浏览量
0
下载量
0
CSCD
- Meta-XML下载
- BibTeX下载
- Endnote下载
- RefMan 下载
- NoteExpress下载
- NoteFirst下载
关联资源
相关文章
相关作者
相关机构
- 技术支持由北京北大方正电子有限公司提供 京ICP备18046208号-2
京公网安备 11010102006710号 - 本系统建议在Chrome、 IE9+ 以上版本浏览器阅读本站内容，360浏览器请切换至极速模式
- Cookies帮助我们提供服务并提供个性化体验。使用本网站，即表示您同意我们使用Cookies
- 总访问量：216460 今日访问量：34