针刀、电针和圆利针对膝骨关节炎模型兔股直肌Bcl-2、Bax、Caspase-3蛋白表达的影响

目的:从细胞凋亡角度探讨不同干预方法对制动致关节损伤的治疗机制。方法:6月龄新西兰兔随机分为正常组、模型组、针刀组、电针组和圆利针组,每组9只,用改良的Videman左后肢伸直位固定制动法建立兔膝关节损伤模型。电针组电针左侧"阳陵泉"-"阴陵泉""内膝眼"-"外膝眼",每次20min,隔日1次,每周3次,共3周;针刀组从兔左侧内、外侧副韧带及髌韧带的中点前缘进针,刀口线与韧带方向平行,分别松解5次,每周1次,共3次;圆利针进针点与针刀组一致,分别松解5次,每周1次,共3次。分别于造模后4、8和12周各时间窗评定各组兔双侧膝关节被动活动范围。采用Western blot法检测各组兔患肢股直肌B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关的X蛋白(Bax)、天冬氨酸特异性半胱氨酸蛋白酶-3(Caspase-3)蛋白表达水平。结果:造模后4周出现稳定的实验性膝骨关节炎兔模型。造模后4、8、12周时,与正常组比较,模型组兔患侧膝关节被动活动度明显降低(P0.01);治疗结束后,与模型组相比,针刀组、电针组和圆利针组兔患侧膝关节被动活动度显著提高(P0.01)。各组患肢股直肌Bcl-2蛋白表达的变化差异无统计学意义(P0.05)。模型组患肢股直肌Bax、Caspase-3蛋白表达较正常组明显升高(P0.05),Bcl-2/Bax比值明显降低(P0.05);针刀组、圆利针组Bax、Caspase-3蛋白表达较模型组明显降低(P0.05),Bcl-2/Bax比值明显升高(P0.05);电针组上述各指标的变化与模型组相比,差异无统计学意义(P0.05)。结论:针刀、圆利针和电针干预膝骨关节炎兔均有治疗效果,针刀和圆利针可通过下调Bax、Caspase-3蛋白表达,升高Bcl-2/Bax比值抑制膝骨关节炎兔股直肌细胞凋亡,从而发挥治疗作用,这与电针治疗关节损伤的机制存在差异。

Effects of Acupotomy,Electroacupuncture or Round-sharp Acupuncture Needle Interventions on Expression of Bcl-2,Bax,Caspase-3 Proteins of Rectus Femoris in Rabbits with Knee Ostarthritis

Objective To observe the effect of acupotomy,electroacupuncture(EA)or round-sharp acupuncture needle intervention on the expression of Bcl-2,Bax and Caspase-3proteins in the rectus femoris in rabbits with knee ostarthritis(KOA), so as to explore their mechanisms underlying improvement of braking-induced joint damage from the cellular apoptosis.Methods Forty-five New Zealand rabbits were equally and randomized into control group,model group,acupotomy(AP)group,EA group and round-sharp acupuncture needle(RSAN)group(n=9in each group).The knee-joint injury model was established by fixing the left knee joint in extention position with plaster bandage.EA(2Hz/100Hz,3mA,20min each time)was applied to the left"Yanglingquan"(GB 34)-"Yinlingquan"(SP 9)and left"Neixiyan"(EX-LE 4)-"Waixiyan"(ST 35)for rabbits in the EA group.The EA treatment was given once daily,3times a week,3weeks in total.For rabbits of the AP group,a needle-knife was held to insert into the front edge of the midpoint,the starting point and the stopping point of the left medial collateral ligamen,lateral collateral ligament and the patellar ligament of the knee to make a loosening manipulation for 5times in a session of treatment,once a week,3times altogether.For rabbits of the RSAN group,a round-sharp needle was performed in the same way to the needle-knife including the stimulation point,the manipulation method and treatment sessions.At the end of the experiment,the left rectus femoris was taken out for detecting the expression of Bcl-2,Bax and Caspase-3proteins with Western blot.ResultsIn comparison with the control group,the passive range of motion(PROM)level was significantly decreased 4,8and 12weeks after modeling(P<0.01),and the expression levels of Bax and Caspase-3proteins in the rectus femoris were considerably upregulated in the model group(P<0.05),while the ratio of Bcl-2/Bax was notably down-regulated(P<0.05)in the model group.Compared with the model group,the PROM level at week 12after modeling in the AP,EA and RSAN groups were significantly increased(P<0.01);while Bax and Caspase-3expression levels in both AP and RSAN groups were considerably downregulated(P<0.05).No significant differences were found among the five groups in Bcl-2expression levels(P>0.05),and between the EA and model groups in Bax and Caspase-3expression levels and the ratio of Bcl-2/Bax(P>0.05).ConclusionAP,RSAN and EA interventions are effective in improving the knee-joint motion range in KOA rabbits,and this effect of both AP and RSAN is closely associated with their actions in lowering the expression of Bax and Caspase-3proteins of the rectus femoris and in raising ratio of Bcl-2/Bax protein(reducing muscular cellular apoptosis).The mechanism of EA intervention in improving PROM may be different.