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冯帅华,吴官保,文哲,姚红艳,姚顺骞,张淼,仇湘中.超微肿痛贴对兔膝骨关节炎模型的软骨中JNK/p38 MAPK信号通路的影响[J].湖南中医药大学学报,2022,42(1):31-36[点击复制] |
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超微肿痛贴对兔膝骨关节炎模型的软骨中JNK/p38 MAPK信号通路的影响 |
冯帅华,吴官保,文哲,姚红艳,姚顺骞,张淼,仇湘中 |
(湖南省中医药研究院附属医院, 湖南 长沙 410006;湖南中医药大学, 湖南 长沙 410208;湘潭市中医院, 湖南 湘潭 411100) |
摘要: |
目的 观察超微肿痛贴对兔膝骨关节炎(knee osteoarthritis,KOA)模型的JNK/p38 MAPK信号通路的mRNA及蛋白表达的影响。方法 将50只3月龄新西兰兔随机分为空白组、模型组、氟比洛芬凝胶贴膏组、超微肿痛贴组,每组各12只,除空白组外,其他3组均采用木瓜蛋白酶膝关节腔注射方式造模。造模成功后,超微肿痛贴组予超微肿痛贴贴于右膝关节,每日1次,每次6 h,连续给药4周。氟比洛芬凝胶贴膏组予氟比洛芬凝胶贴膏贴于右膝关节,每日1次,每次6 h,连续给药4周。干预4周后空气栓塞法处死所有实验兔,观察各组关节软骨Mankin’s评分;ELISA法检测关节液中TNF-α水平;采用RT-PCR检测关节软骨细胞中JNK和p38 MAPK mRNA的相对表达量;Western blot检测软骨中p38 MAPK、JNK、MMP-13蛋白表达。结果 超微肿痛贴组与氟比洛芬凝胶贴膏组关节软骨Mankin’s评分及关节液中TNF-α表达水平显著低于模型组(P<0.05),并且JNK和p38 MAPK mRNA的表达水平,及p38 MAPK、JNK、MMP-13蛋白表达水平较模型组均显著下降(P<0.05),超微肿痛贴组各指标与氟比洛芬凝胶贴膏组相比,差异均无统计学意义(P>0.05)。结论 超微肿痛贴能够有效减轻兔KOA的软骨损伤,调控关节软骨JNK/p38 MAPK信号通路,抑制软骨MMP-13表达,减少关节液中TNF-α表达水平,这可能为其治疗KOA的部分作用机制。 |
关键词: 膝骨关节炎 超微肿痛贴 关节软骨 MAPK信号通路 软骨损伤 |
DOI:10.3969/j.issn.1674-070X.2022.01.007 |
投稿时间:2021-04-05 |
基金项目:国家中医药管理局全国名老中医药专家仇湘中传承工作室建设项目(国中医药人教〔2016〕42号);湖南省自然科学基金面上项目(2018JJ2235);湖南省教育厅科学研究项目(17C1218)。 |
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Effect of Chaowei Zhongtong Patch on JNK/p38 MAPK signaling pathway in cartilage of rabbit knee osteoarthritis model |
FENG Shuaihua,WU Guanbao,WEN Zhe,YAO Hongyan,YAO Shunqian,ZHANG Miao,QIU Xiangzhong |
(Affilated Hospital of Hunan Academy of Chinese Medicine, Changsha, Hunan 410006, China;Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Xiangtan Hospital of Traditional Chinese Medicine, Xiangtan, Hunan 411100, China) |
Abstract: |
Objective To observe the effect of Chaowei Zhongtong Patch on the mRNA and protein expression of JNK/p38 MAPK signaling pathway in rabbit knee osteoarthritis (KOA) model. Methods A total of 50 3-month old New Zealand rabbits were randomly divided into blank group, model group, flurbiprofen cataplasm group and Chaowei Zhongtong Patch group, with 12 rabbits in each group; except for the blank group, the other three groups were injected with papain into knee joint cavity for modeling. After the successful modeling, the Chaowei Zhongtong Patch group was applied Chaowei Zhongtong Patch to the right knee joint, once a day, for 6 h each time, for 4 weeks. Flurbiprofen cataplasm group was given flurbiprofen gel patch on right knee joint, once a day, 6 h each time, for 4 weeks. After 4 weeks of intervention, all rabbits were killed by air embolism, then the Mankin's score of articular cartilage of each group was observed; TNF-α level in synovial fluid was detected by ELISA; relative expression levels of JNK and p38 MAPK mRNA in articular chondrocytes were tested by RT-PCR; the protein expression levels of p38 MAPK, JNK and MMP-13 in articular cartilage were detected by Western blot. Results The Mankin's score of articular cartilage and the expression level of TNF-α in synovial fluid of the Chaowei Zhongtong Patch group were significantly lower than those in the model group (P<0.05), and the expression levels of JNK and p38 MAPK mRNA and the protein expression levels of p38 MAPK, JNK and MMP-13 of the Chaowei Zhongtong Patch group were remarkably declined compared with those in the model group (P<0.05), there were no statistical significances between Chaowei Zhongtong Patch group and flurbiprofen cataplasm group (P>0.05). Conclusion Chaowei Zhongtong Patch can effectively reduce the cartilage damage of rabbit KOA, regulate JNK/p38 MAPK signaling pathway, inhibit the expression levels of MMP-13 in cartilage, and decrease the expression level of TNF-α in synovial fluid, which may be part of its acting mechanisms in the treatment of knee KOA. |
Key words: knee osteoarthritis Chaowei Zhongtong Patch joint cartilages cartilage damage |
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