目的:观察补阳还五汤联合丰富环境对血管痴呆大鼠学习记忆,半胱氨酸天冬氨酸特异性蛋白酶3(Caspase-3)和凋亡相关蛋白Fas,Bcl-2相关蛋白X(Bax)表达的影响。方法:采用线栓法阻断大鼠大脑中动脉,制备MCAO血管痴呆大鼠模型,分别给以补阳还五汤为16.1 g·kg-1ig、丰富环境干预及补阳还五汤16.1 g·kg-1ig联合丰富环境的不同处理,连续治疗30 d后取材,采用Morris水迷宫检测学习记忆功能,酶联免疫法检测额叶皮层Caspase-3含量,免疫组织化学染色法结合图像分析技术测定Fas、Bax的表达。结果:补阳还五汤组、丰富环境可改善血管痴呆大鼠空间参考记忆能力(P〈0.01),联合治疗可改善模型大鼠空间参考记忆及工作记忆(P〈0.01或P〈0.05);补阳还五汤治疗能明显减少额叶皮层凋亡基因Fas、Bax表达(P〈0.01或P〈0.05),丰富环境干预也可明显降低额叶皮层Bax蛋白表达(P〈0.01);经联合治疗后,大鼠额叶皮层Fas表达较丰富环境明显减少(P〈0.05),Bax表达较补阳还五汤单独治疗下调(P〈0.05),而Caspase-3活性则较补阳还五汤、丰富环境单独处理大鼠均明显降低(P〈0.01)。结论:补阳还五汤联合丰富环境具有协同增效的作用,改善血管痴呆大鼠学习记忆,下调Fas、Bax凋亡蛋白的表达,抑制Caspase-3的活性。
Objective: To investigate the effects of Buyang Huanwu decoction(BYHWD) combined with enriched environment on learning memory function and the expressions of apoptosis-related proteins Fas,Bax and Caspase-3 in rats with vascular dementia.Method: The animal model of middle cerebral artery(MCA) ischemia was adopted.The BYHWD group,enriched environment group and combing group were given relative treatment.The rats were killed after thirty days continuous treating.Learning and memory ability of animal were evaluated by Morris water maze method.The ELISA method was used to measure the levels of Caspase-3 in the frontal cortex.The expression of Fas and Bax were observed by immunohistochemical staining and then analyzed by imaged analysis system.Result: The BYHWD group and enriched environment group could improve the spatial reference memory ability(P0.01),BYHWD combined with enriched environment could improve the spatial reference memory and working memory ability of rats(P0.01 or P0.05);YHWD treatment could reduce the expression of Fas and Bax in the frontal cortex(P0.01 or P0.05),enriched environment intervention also could reduce the expression of Bax(P0.01);after the treatment of BYHWD combined with enriched environment,the expression of Fas in the frontal cortex was significantly decreased compared with enriched environment group(P0.05),the expression of Bax was markedly decreased compared with the BYHWD group(P0.05),the expression of Caspase-3 was significantly decreased compared with the BYHWD group and enriched environment group(P0.01).Conclusion: BYHWD combined with enriched environment had synergistic effect that it could improve the ability of learning memory of rats with vascular dementia,reduce the expression of Fas,Bax and inhibite the activation of Caspase-3.