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[针刀对膝骨关节炎家兔软骨组织中细胞衰老标志物p16Ink4a和p21Waf1/Cip1表达的影响]

[Effect of acupotomy on expressions of cellular senescence markers p16Ink4a and p21Waf1/Cip1 in cartilage tissue of rabbits with knee osteoarthritis].

作者信息

Pan Jun-Kang, Zhang Yi, Chen Xi-Lin, Xing Long-Fei, Xu Yue, He Meng, Hu Rui, Li Jia-Hui, Guo Chang-Qing, Guo Yan

机构信息

School of Acupuncture-Moxibustion and Tuina, Beijing University of Chinese Medicine, Beijing 100029, China.

School of Acupuncture-Moxibustion and Tuina, Beijing University of Chinese Medicine, Beijing 100029, China; Kunming Municipal Hospital of Traditional Chinese Medicine, Kunming 650500.

出版信息

Zhen Ci Yan Jiu. 2023 Jul 25;48(7):658-65. doi: 10.13702/j.1000-0607.20221091.

Abstract

OBJECTIVE

To observe the effect of acupotomy on the expressions of p16 and p21 in knee osteoarthritis (KOA) rabbits,so as to analyze whether acupotomy can treat KOA by inhibiting the cellular senescence of chondrocytes.

METHODS

Twenty-four New Zealand male rabbits were randomly divided into normal, model, acupotomy and electroacupuncture (EA) groups, with 6 rabbits in each group. The KOA model was established by left hindlimb straightening fixation. After modeling, rabbits in the acupotomy group were treated with acupotomy loosening therapy on high stress points around the affected knee joints such as tendons attachment points of vastus medialis, vastus lateralis, rectus femoris, biceps femoris and pes anserine bursa, once a week for 3 weeks. In the EA group, "Xuehai"(SP10), "Liangqiu" (ST34),"Neixiyan" (EX-LE4) and "Waixiyan" (ST35) on the affected hindlimb were selected for EA treatment (3 mA, 2 Hz/100 Hz), 20 min each time, once every other day for 3 weeks. Before and after treatments, the knee Lequesne MG score and passive range of motion (PROM) of the affected knee joint were evaluated. After the treatments, the expressions of p16 and p21 in the cartilage tissue of the affected knee joint were detected by immunohistochemistry and Western blot respectively.

RESULTS

Before and after treatment, compared with the normal group, the Lequesne MG score was significantly increased (<0.01), the PROM was significantly decreased (<0.01) in the model group. After treatment, compared with the normal group, the positive expression and protein expression levels of p16 and p21 were significantly increased (<0.01) in the model group; compared with the model group, the Lequesne MG score was significantly decreased (<0.01), the PROM was significantly increased (<0.01), the positive expression and protein expression levels of p16 and p21 were significantly decreased (<0.01,<0.05) in the acupo-tomy and EA groups; compared with the EA group, the Lequesne MG score was decreased (<0.05), the PROM was increased (<0.05), the positive expression and protein expression levels of p16 and p21 were decreased (<0.05,<0.01) in the acupotomy group.

CONCLUSION

Acupotomy intervention can down-regulate the expressions of cellular senescence markers p16 and p21 in chondrocytes, indicating that acupotomy therapy may alleviate cartilage degeneration by inhibiting chondrocyte premature cellular senescence to treat KOA.

摘要

目的

观察针刀对膝骨关节炎(KOA)兔p16和p21表达的影响,分析针刀是否可通过抑制软骨细胞衰老来治疗KOA。

方法

将24只新西兰雄性兔随机分为正常组、模型组、针刀组和电针组,每组6只。采用左后肢伸直固定法建立KOA模型。造模后,针刀组对患侧膝关节周围高应力点如股内侧肌、股外侧肌、股直肌、股二头肌肌腱附着点及鹅足囊进行针刀松解治疗,每周1次,共3周。电针组选取患侧后肢的“血海”(SP10)、“梁丘”(ST34)、“内膝眼”(EX-LE4)和“外膝眼”(ST35)进行电针治疗(3 mA,2 Hz/100 Hz),每次20分钟,隔日1次,共3周。治疗前后分别评估患侧膝关节Lequesne MG评分及被动活动度(PROM)。治疗后,分别采用免疫组化和Western blot检测患侧膝关节软骨组织中p16和p21的表达。

结果

治疗前后,与正常组比较,模型组Lequesne MG评分显著升高(<0.01),PROM显著降低(<0.01)。治疗后,与正常组比较,模型组p16和p21的阳性表达及蛋白表达水平显著升高(<0.01);与模型组比较,针刀组和电针组Lequesne MG评分显著降低(<0.01),PROM显著升高(<0.01),p16和p21的阳性表达及蛋白表达水平显著降低(<0.01,<0.05);与电针组比较,针刀组Lequesne MG评分降低(<0.05),PROM升高(<0.05),p16和p21的阳性表达及蛋白表达水平降低(<0.05,<0.01)。

结论

针刀干预可下调软骨细胞中细胞衰老标志物p16和p21的表达,表明针刀疗法可能通过抑制软骨细胞过早衰老来减轻软骨退变,从而治疗KOA。

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