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

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

Objective: To observe the effect of acupotomy on the expressions of p16Ink4a and p21Waf1/Cip1 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 p16Ink4a and p21Waf1/Cip1 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 (P<0.01), the PROM was significantly decreased (P<0.01) in the model group. After treatment, compared with the normal group, the positive expression and protein expression levels of p16Ink4a and p21Waf1/Cip1 were significantly increased (P<0.01) in the model group; compared with the model group, the Lequesne MG score was significantly decreased (P<0.01), the PROM was significantly increased (P<0.01), the positive expression and protein expression levels of p16Ink4a and p21Waf1/Cip1 were significantly decreased (P<0.01,P<0.05) in the acupo-tomy and EA groups; compared with the EA group, the Lequesne MG score was decreased (P<0.05), the PROM was increased (P<0.05), the positive expression and protein expression levels of p16Ink4a and p21Waf1/Cip1 were decreased (P<0.05,P<0.01) in the acupotomy group.

Conclusion: Acupotomy intervention can down-regulate the expressions of cellular senescence markers p16Ink4a and p21Waf1/Cip1 in chondrocytes, indicating that acupotomy therapy may alleviate cartilage degeneration by inhibiting chondrocyte premature cellular senescence to treat KOA.

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[针刺对膝骨关节炎兔软骨组织中细胞衰老标志物p16Ink4a和p21Waf1/Cip1表达的影响]。
目的:观察针刺对膝骨关节炎(KOA)兔p16Ink4a和p21Waf1/Cip1表达的影响,探讨针刺是否能通过抑制软骨细胞衰老来治疗KOA。方法:24只新西兰雄兔随机分为正常组、模型组、针刺组和电针组,每组6只。采用左后肢伸直固定法建立KOA模型。造模后,针刀组家兔对患膝关节周围的高应力点,如股内侧肌、股外侧肌、股直肌、股二头肌和筋膜囊的肌腱附着点进行针刀松动治疗,每周1次,持续3周。电针组选用患肢“血海”、“凉秋”、“内息炎”、“外息炎”进行电针治疗(3mA,2Hz/100Hz),每次20min,隔日1次,疗程3周。在治疗前后,评估受影响膝关节的Lequesne MG评分和被动活动范围(PROM)。治疗后,分别用免疫组织化学和Western印迹法检测p16Ink4a和p21Waf1/Cip1在膝关节软骨组织中的表达。结果:治疗前后与正常组比较,Lequesne MG评分显著升高(PPInk4a和p21Waf1/Cip1显著升高(PPPInk4a和p21Waf1/Cip显著降低(PPPPInk4a、p21Waf1/cip降低(PPC结论:针刀治疗可下调软骨细胞中细胞衰老标志物p16Ink4a和p21Waf1/Cip1的表达,提示针刀治疗可以通过抑制软骨细胞早衰来缓解软骨变性。
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