复方益糖康联合运动训练对糖尿病脑梗死大鼠神经功能修复作用

Effect of Compound Yitangkang(复方益糖康)Combined with Exercise Training on Neurological Function of Diabetic Cerebral Infarction Rats

目的观察复方益糖康联合运动训练对糖尿病脑梗死大鼠神经功能改善,成熟少突胶质细胞标志物(APC)、髓鞘碱性蛋白(MBP)、半胱氨酸-天冬氨酸蛋白酶-3(Caspase-3)表达的影响,探究复方益糖康联合运动训练对糖尿病脑梗死大鼠神经功能修复作用。方法160只成年雄性SD大鼠随机分为假手术组(n=15)、模型组(n=45)、中药组(n=35)、运动组(n=35)、联合组(n=30)。先后行糖尿病造模、大鼠大脑中动脉永久性闭塞术。中药组、联合组进行中药灌服,每日1次;运动组、联合组进行电动跑台训练,每日1次,每次30 min;假手术组和模型组不予中药、运动训练;上述各组共治疗3周。分别于术后24 h、7 d、21 d进行大鼠神经功能mNSS评分,免疫荧光法检测APC、MBP、Caspase-3的变化,用TUNEL法检测细胞凋亡,免疫荧光双标检测APC/Caspase-3共表达情况。用Western blot检测脑梗死缺血半暗带区脑组织中APC、MBP、Caspase-3蛋白的表达。结果缺血后24 h、7 d、21 d,神经功能行为学评分、TUNEL染色、Caspase-3免疫荧光和蛋白表达:与模型组比较,中药组、运动组、联合组大鼠明显降低(P<0.05),与单一治疗组比较,联合组显著升高(P<0.01)。少突胶质细胞、髓鞘阳性细胞及蛋白表达:与模型组比较,中药组、运动组、联合组大鼠明显升高(P<0.05),与单一治疗组比较,联合组显著升高(P<0.01)。结论益糖康联合运动训练对糖尿病脑梗死大鼠的神经功能修复,上调APC、MBP蛋白表达,下调Caspase-3蛋白表达,可能与促进成熟少突胶质细胞增多、髓鞘稳定,抑制成熟少突胶质细胞凋亡有关。

Objective To observe the effect of Compound Yitangkang(复方益糖康)combined with exercise training on the improvement of nerve function and the expression of APC,MBP and Caspase-3 in diabetic cerebral infarction rats,and to explore the effect of Compound Yitangkang combined with exercise training on the repair of nerve function in diabetic cerebral infarction rats.Methods 160 adult male SD rats were randomly divided into sham operation group(n=15),model group(n=45),traditional Chinese medicine group(n=35),exercise group(n=35)and combined group(n=30).Diabetic model was established and permanent occlusion of middle cerebral artery was performed in rats.Chinese medicine group and combined group were given Chinese medicine once a day;The exercise group and the joint group were trained on electric treadmill,once a day,30 minutes each time;The sham operation group and the model group were not treated with traditional Chinese medicine and exercise.All groups were treated for 3 weeks.The changes of mature oligodendrocyte(APC),myelin sheath(MBP)and Caspase-3 were measured by mNSS score and immunofluorescence at 24 h,7 d and 21 d after operation.Apoptosis was detected by TUNEL method,and APC/Caspase-3 co-expression was detected by immunofluorescence double labeling.Western blot was used to detect the expression of APC,MBP and Caspase-3 protein in ischemic penumbra of cerebral infarction.Results After 24 hours,7 days and 21 days of ischemia,neurological behavioral score,TUNEL staining,Caspase-3 immunofluorescence and protein expression:Compared with the model group,the rats in the Chinese medicine group,the exercise group and the combined group decreased significantly(P<0.05),while those in the combined group increased significantly(P<0.01).The expression of oligodendrocytes,myelin sheath positive cells and protein:Compared with the model group,the rats in the Chinese medicine group,exercise group and combination group increased significantly(P<0.05),and compared with the single treatment,the combination group increased significantly(P<0.01).Conclusion Yitangkang combined with exercise training can improve the neurological function of diabetic cerebral infarction rats,which may be related to up-regulating APC and MBP protein expression,downregulating Caspase-3 protein expression,inhibiting oligodendrocyte apoptosis and promoting myelin sheath stability.