针药联合对阿尔兹海默病模型大鼠小脑皮质谷氨酸受体2及Caspase-3表达的影响

Effects of electroacupuncture-gastrodin combined on the expressions of glutamate receptor 2 and Caspase-3 in the cerebellar cortex of rats with Alzheimer’s disease

目的观察针药联合对阿尔兹海默病(AD)模型大鼠小脑皮质谷氨酸受体(GluR2)及天冬氨酸蛋白水解酶(Caspase-3)表达的影响,探讨小脑在AD发病机制中的作用。方法30只雄性SD大鼠随机分为5组:对照组(C组)、模型组(M组)、电针组(EA组)、天麻素组(Gas组)、针药联合组(EA+Gas组),每组6只。C组不作任何处理;M组大鼠给予双侧海马内微量注射5μL的Aβ1-42进行造模;EA组予以频率2 Hz、强度1mA的电针刺激,选取穴位“大椎”“百会”“足三里”,每日30 min,连续4周。Gas组予以腹腔注射Gas注射液(10 mg/kg),每天1次,连续4周;EA+Gas组同时予以电针和Gas治疗,连续4周。Morris水迷宫检测大鼠学习记忆行为能力,Nissl染色和免疫组化方法观察各组大鼠小脑皮质细胞形态和数量,Western Blot法检测各组GluR2和Caspase-3蛋白的表达。结果M组小脑皮质中浦肯野层细胞数量减少(P<0.05),GluR2表达降低(P<0.05),Caspase-3表达增高(P<0.05);各治疗组较M组的细胞形态趋于正常,GluR2的表达增加(P<0.05),Caspase-3的表达降低(P<0.05),其中以EA+Gas组改变最为明显。结论电针、天麻素及针药联合均可通过调节GluR2和Caspase-3的表达水平进而改善AD模型大鼠学习记忆能力,其中针药联合效果最佳。

Objective To observe the effects of electroacupuncture combined with medicine on the expressions of glutamate receptor 2(GluR2)and Caspase-3 in the cerebellar cortex of rats with Alzheimer’s disease(AD)and explore the role of cerebellum in the pathogenesis of AD.Methods Thirty male SD rats were randomly divided into 5 groups:control group(C group),model group(M group),electroacupuncture group(EA group),gastrodin group(Gas group),and acupuncture-medication group(EA+Gas group),with 6 rats in each group.The C group received no treatment.The M group was modeled by microinjection of 5μL Aβ1-42 into bilateral hippocampus.The EA group received electroacupuncture stimulation at a frequency of 2 Hz and an intensity of 1 mA at acupoints“Da Zhui”,“Baihui”,and“Zu San Li”for 30 minutes daily for four consecutive weeks.The Gas group received intraperitoneal injection of gastrodin solution(10 mg/kg)once a day for four weeks.The EA+GA group was treated with EA and Gas for 4 consecutive weeks.Morris Water Maze was used to test the learning and memory behavior of rats;Nissl staining and immunohistochemistry were performed to observe the morphology and quantity of cerebellar cortex cells in each group,and the expressions of GluR2 and Caspase-3 proteins were detected by Western Blot.Results In the M group,the number of Purkinje cells decreased in the cerebellar cortex(P<0.05),the expression of GluR2 decreased(P<0.05),and Caspase-3 increased(P<0.05).The cell morphology of each treatment group tended to be normal compared with the M group,with increased GluR2 expression(P<0.05)and decreased Caspase-3 expression(P<0.05).The EA+Gas group showed the most significant changes.Conclusion The EA,gastrodin and EA+Gas can improve the learning and memory ability of AD model rats by regulating the expression levels of GluR2 and Caspase-3,and the combination of two methods has the best efficacy.