健脾益智汤对阿尔茨海默病大鼠行为学影响及脑内尿素代谢机制研究

Study of Jianpi Yizhi Decoction on the effects of behavioral and mechanism of brain urea metabolism of Alzheimer’s disease rats

目的:观察健脾益智汤对阿尔茨海默病(AD)模型大鼠的学习记忆能力、脑内尿素含量、相关尿素转运蛋白及基因的影响。方法:90只SPF级雄性SD大鼠随机分为对照组、假手术组、模型组、健脾益智汤组、吡拉西坦组,每组18只。采用海马区注射Aβ_(1-40)制备AD动物模型。各组术后第2周起开始灌胃,分别持续7、14、28 d后进行Morris水迷宫实验,采用脲酶法观察和分析各组大鼠脑内尿素含量,实时荧光定量PCR法检测SLC14A1m RNA表达情况,Western blot法检测尿素通道蛋白B(UTB)蛋白含量。结果:治疗7 d开始,与模型组比较,健脾益智汤组及吡拉西坦组逃避潜伏期及脑内尿素含量均显著减少(P<0.01,P<0.05)。治疗14、28 d,与模型组比较,健脾益智汤组及吡拉西坦组穿越平台次数显著增加(P<0.05,P<0.01),SLC14A1 mRNA表达水平和UTB蛋白含量显著升高(P<0.05,P<0.01)。结论:健脾益智汤可能通过上调SLC14A1基因和UTB蛋白表达,提高尿素转运能力,达到改善学习记忆障碍的作用。

Objective: To observe the effects of Jianpi Yizhi Decoction on learning and memory ability, brain urea content, related urea transporters and genes of Alzheimer’s disease(AD) model rats. Methods: Ninety SPF male SD rats were randomly divided into control group, sham operation group, model group, Jianpi Yizhi Decoction group and piracetam group,18 rats in each group. AD experimental animal model was prepared by Aβ_(1-40) was injected into hippocampus. Each group began to gavage from the second week after operation. Morris water maze test was carried out after 7 days, 14 days and 28days respectively. The urea content in the brain of rats in each group was observed and analyzed by Urease method, the mRNA expression oF SLC14A1 gene was detected by real-time fluorescence quantitative PCR, and the UTB protein content was detected by Western blot. Results: Since 7 days of treatment, compared with the model group, the escape latency and the content of urea in brain of rat in Jianpi Yizhi Decoction group and Piracetam group were decreased(P<0.01, P<0.05). After 14, 28 days of treatment,compared with model group, the number of crossing the platform in Jianpi Yizhi Decoction group and Piracetam group were decreased increased(P<0.05, P<0.01), and the expression levels of SLC14A1 mRNA and UTB protein were increased(P<0.05,P<0.01). Conclusion: Jianpi Yizhi Decoction may improve learning and memory impairment by up regulating the expression of SLC14A1 gene and UTB protein and improving the ability of urea transport.