加减薯蓣丸对AD模型大鼠行为学及海马TNF-α、Caspase-3表达的影响

Effects of Modified Shuyu Pill on the Behavior,Expression of TNF-α and Caspase-3 in Hippocampus of AD Model Rats

目的观察加减薯蓣丸对阿尔茨海默病(AD)模型大鼠行为学及海马肿瘤坏死因子-α(TNF-α)、半胱氨酸蛋白酶-3(Caspase-3)表达的影响。方法将SPF级雄性SD大鼠,侧脑室注射A 1-42寡聚体法建模。建模成功后随机分为模型组、中药组及西药组,设假手术组对照,每组10只。模型组、假手术组予以生理盐水10mL/(kg·d)灌胃,中药组予以加减薯蓣丸10g/(kg·d)灌胃,西药组予以多奈哌齐0.45mg/(kg·d)灌胃,连续4周。Morris水迷宫观察大鼠学习记忆能力,Western Blot法检测大鼠海马TNF-α、Caspase-3蛋白表达,实时荧光定量PCR法检测大鼠海马TNF-α、Caspase-3 mRNA表达。结果第6天试验中,与假手术组相比,模型组逃避潜伏期延长(P<0.01);与模型组相比,中药组、西药组逃避潜伏期缩短(P<0.01)。空间探索试验中,与假手术组相比,模型组穿越平台次数和原平台象限停留时间比减少(P<0.01);与模型组相比,中药组、西药组穿越平台次数和原平台象限停留时间比增加(P<0.01)。与假手术组相比,模型组TNF-α、Caspase-3蛋白和m RNA的表达升高(P<0.01);与模型组相比,中药组和西药组TNF-α、Caspase-3蛋白和mRNA的表达降低(P<0.05,P<0.01);与西药组比较,中药组TNF-α、Caspase-3蛋白及TNF-αmRNA表达降低更为明显(P<0.05,P<0.01)。结论加减薯蓣丸可改善AD模型大鼠学习记忆能力,其机制可能与抑制海马TNF-α、Caspase-3表达有关。

Objective To observe the effects of Modified Shuyu Pill on the behavior,expression of tumor necrosis factor-α(TNF-α)and Caspase-3 in hippocampus of Alzheimer’s disease(AD)model rats. Methods SPF-grade male SD rats were injected into lateral ventricle using A 1-42 oligomer to establish the animal model of AD. Rats with successful modeling were randomly divided into model group,TCM group and western medicine group,and the sham operation group was set up as control,with 10 rats in each group. The model group and sham operation group were given normal saline 10 m L/(kg·d)intragastrically daily,TCM group was given MSP 10 g/(kg·d)intragastrically daily,while the western medicine group was given donepezil 0.45 mg/(kg·d)intragastrically daily. The therapeutic course was 4 weeks. Morris water maze was used to observe the learning and memory ability of rats. Western Blot was used to detect the protein expression of TNF-αand Caspase-3 in hippocampus of rats. The expressions of TNF-αand Caspase-3 m RNA in hippocampus of rats were detected by real-time fluorescence quantitative PCR. Results The escape latency was prolonged in model group compared with sham group(P<0.01)on the 6 th day. Compared with model group,the escape latency of TCM group and western medicine group were shortened(P<0.01);in the space exploration experiment,compared with the sham operation group,the number of times the model group crossed the platform and the ratio of residence time in the original platform quadrant were reduced(P<0.01);compared with model group,the times of crossing the platform and the ratio of residence time in the original quadrant of TCM group and western medicine group were increased(P<0.01);compared with sham operation group,the expressions of TNF-α,Caspase-3 protein and m RNA in model group were significantly increased(P<0.01);compared with model group,the expressions of TNF-α,Caspase-3 protein and m RNA in TCM group and western medicine group were significantly decreased(P<0.05,P<0.01);compared with western medicine group,the expression of TNF-α,Caspase-3 protein and TNF-α mRNA in TCM group decreased significantly(P<0.05,P<0.01). Conclusion Modified Shuyu Pill can effectively improve the learning and memory ability of AD model rats,and the mechanism may be related to the inhibition of TNF-α and Caspase-3 expression in hippocampus.