针刀干预对膝骨关节炎模型兔股直肌p16、p21表达的影响

Effects of Acupotomy Intervention on Expressions of p16 and p21 inSkeletal Muscle Cells of Rabbits with KOA

目的:观察针刀治疗后膝骨关节炎(KOA)兔股直肌内p16、p21蛋白表达的变化,探讨针刀治疗对KOA兔股直肌细胞衰老的抑制作用。方法:清洁级新西兰雄兔24只随机分为针刀组、电针组、正常组和模型组,每组6只。采用改良Videman左后肢膝关节制动制备KOA兔模型。针刀组和电针组各干预3周后,用改良Lequesne MG指数评分和膝关节被动活动度(PROM)进行行为学评价,HE染色法观察光镜下股直肌肌肉及肌腱病理学变化并用Western blot法检测股直肌中p21、p16的蛋白表达。结果:与正常组相比,模型组兔Lequesne MG评分显著升高,PROM显著降低(P<0.01);光镜下股直肌肌束排列紊乱,有炎细胞浸润,肌纤维横截面积显著减少(P<0.01),肌细胞数量显著增多(P<0.01);p16和p21的蛋白表达上调(P<0.01,P<0.05)。与模型组相比,针刀组与电针组Lequesne MG评分显著降低,PROM显著升高(P<0.01);光镜下见肌细胞再生,炎细胞浸润减轻,肌纤维横截面积显著增多,肌细胞数量显著减少(P<0.01)。p16和p21蛋白均呈不同程度降低(P<0.01,P<0.05)。结论:针刀干预可改善膝关节周围骨骼肌力学性能,促进KOA的恢复,其机制可能与下调p16、p21表达和抑制股直肌细胞衰老以修复萎缩的股直肌有关。

Objective:To observe the protein expressions of p16 and p21 in the rectus femoris of rabbits with knee osteoarthritis(KOA)after acupotomy therapy,and to analyze the inhibiting effect of acupotomy on the senescence of the rectus femoris cells.Methods:24 SPF New Zealand male rabbits were randomly divided into the acupotomy group,the electro-acupuncture(EA)group,the normal group and the model group,with 6 rabbits in each group.KOA model was prepared with the modified Videman left hind leg immobilization method.After 3 weeks of intervention,the modified Lequesne MG score and the passive range of motion(PROM)were used for behavioral evaluation.Pathological changes of rectus femoris and its tendon were observed by HE staining and the protein expressions of p16 and p21 in the rectus femoris were detected by Western blot method.Results:Compared with those in the normal group,the Lequesne MG score was significantly increased and the PROM was greatly decreased(P<0.01);the fascicles of rectus femoris were disorganized with inflammatory cell infiltration;the cross-sectional area(CSA)of muscle fibers was decreased significantly(P<0.01);the number of muscle fibers was increased significantly(P<0.01);the expressions of p16 and p21 were increased significantly in the model group(P<0.01,P<0.05).Compared with those in the model group,the Lequesne MG score was decreased significantly and the PROM was increased significantly(P<0.01);the fascicles of rectus femoris regenerated with decreased inflammatory cells;the CSA was increased significantly and the number of muscle fibers was decreased significantly(P<0.01);the expressions of p16 and p21 were decreased significantly in the acupotomy group and the EA group(P<0.01,P<0.05).Conclusion:Acupotomy intervention can improve the mechanical properties of skeletal muscles around the knee joint and promote the recovery of KOA,which mechanism may be related to down-regulating the expressions of p16 and p21 in skeletal muscle cells,inhibiting the senescence process of the rectus femoris cells and ameliorating repair of mucle cells in rabbits with KOA.