针康法对脑缺血大鼠缺血半暗区细胞凋亡及X连锁凋亡抑制蛋白和cleaved-caspase-9蛋白表达的影响

摘要

Objective To explore the effect of acupuncture-rehabilitation therapy on apoptosis and the expression of X-linked inhibitor of apoptosis protein (XIAP) and cleaved-caspase-9 in ischemic penumbra of rats with cerebral ischemia. Methods A total of 180 male Sprague-Dawley rats were randomly divided into sham operation group,model group,acupuncture group,rehabilitation group and acupunc-ture-rehabilitation therapy group.Each group was divided into three days,seven days and 14 days subgroups(n=12).The model was estab-lished with the modified Longa suture method. The model group and the sham operation group received no treatment. The acupuncture group received cluster needling of scalp acupuncture,the rehabilitation group received treadmill training,and the acupuncture-rehabilitation therapy group received both acupuncture and treadmill training.On the third,seventh and 14th days after modeling,the neurological func-tion was assessed with modified Neurological Severity Score(mNSS).The apoptosis was observed by TUNEL staining and the expression of XIAP and cleaved-caspase-9 protein was determined by Western blotting in cerebral ischemic penumbra,respectively.Results Compared with the model group,the mNSS scores decreased,the rate of apoptosis decreased,the expression of XIAP protein increased,and the expres-sion of cleaved-caspase-9 decreased in each treatment group at each time point(P<0.05).Compared with the other two treatment groups,the rate of apoptosis further decreased at each time points(P<0.05),the expression of XIAP protein further increased at each time points(P<0.05),the mNSS score further decreased(P<0.05)and the expression of cleaved-caspase-9 protein further decreased(P<0.05)seven days and 14 days after modeling in the acupuncture-rehabilitation therapy group.Conclusion Acupuncture and rehabilitation therapy could re-duce the neurological deficit in rats with cerebral ischemia,which was superior to the simple acupuncture treatment and rehabilitation train-ing.The mechanism may be related to th reduction of apoptosis by promoting XIAP protein expression and inhibiting caspase-9 protein acti-vation.%目的 观察针康法对脑缺血大鼠缺血半暗区细胞凋亡及X连锁凋亡抑制蛋白(XIAP)和cleaved-caspase-9蛋白表达的影响.方法 选取180只雄性Sprague-Dawley大鼠,通过随机数字表法随机分为假手术组、模型组、针刺组、康复组和针康组.每组再分为3 d、7 d和14 d三个亚组(n=12).采用改良Longa线栓法进行造模,假手术组和模型组不予治疗,针刺组进行头穴丛刺治疗,康复组进行跑台康复训练,针康组进行针康法治疗.术后各时间点对大鼠进行改良神经功能缺损评分(mNSS)后取材,采用TUNEL染色观察大鼠脑缺血半暗区细胞凋亡率,Western blotting检测脑缺血半暗区XIAP和cleaved-caspase-9蛋白表达.结果 术后各时间点,与模型组比较,各治疗组mNSS评分均降低(P<0.05),细胞凋亡率均降低(P<0.05),XIAP蛋白表达上调(P<0.05), cleaved-caspase-9蛋白表达下调(P<0.05);与针刺组、康复组比较,针康组细胞凋亡率进一步降低(P<0.05),XIAP蛋白表达进一步上调(P<0.05).术后7 d、14 d,针康组mNSS评分进一步降低(P<0.05),针康组cleaved-caspase-9蛋白表达进一步下调(P<0.05).结论 针康法能降低脑缺血大鼠神经功能缺损,优于单独的头穴丛刺和康复训练,其机制可能与促进XIAP蛋白表达,抑制caspase-9蛋白活化,从而减少细胞凋亡有关.