黄蒲通窍胶囊对阿尔茨海默病模型大鼠Wnt/Ca^(2+)信号通路的影响

Effect of Huangpu Tongqiao Capsule on the Wnt/Ca^(2+) Signaling Pathway in a Rat Model of Alzheimer's Disease

目的观察黄蒲通窍胶囊对阿尔茨海默病(Alzheimer's disease,AD)模型大鼠学习记忆的影响,探究其潜在作用机制。方法将30只SD大鼠随机分为空白组、模型组、黄蒲通窍胶囊组,每组10只,腹腔注射D-半乳糖联合双侧海马注射Aβ_(25-35)复制AD大鼠模型;给药28 d,采用Morris水迷宫实验检测大鼠的学习记忆能力,检测后取材,采用苏木精-伊红染色观察海马神经细胞损伤情况,采用Western blot法检测海马组织中Wnt5、Frizzled、PLCβ蛋白表达水平,采用ELISA法检测血清三磷酸肌醇(inositol triphosphate,IP3)表达水平。结果Morris水迷宫实验结果显示:训练期,各组大鼠的逃避潜伏期随时间的增加呈下降趋势;测试期,与空白组比较,模型组大鼠逃避潜伏期明显延长,目标象限路程与时间百分比明显减少(P<0.05);与模型组比较,黄蒲通窍胶囊组大鼠逃避潜伏期明显缩短,目标象限路程百分比和时间百分比明显增加(P<0.05)。苏木精-伊红染色结果显示:与空白组比较,模型组大鼠海马神经细胞萎缩破裂、层数减少、排列松散;与模型组比较,黄蒲通窍胶囊组大鼠海马神经细胞损伤减轻。Western blot法检测结果显示:与空白组比较,模型大鼠Wnt5、Frizzled、PLCβ表达水平均明显升高(P<0.05);与模型组比较,黄蒲通窍胶囊组大鼠Wnt5、Frizzled、PLCβ表达水平均明显降低(P<0.05)。ELISA法检测结果显示:与空白组比较,模型组大鼠IP3表达水平明显升高(P<0.05);与模型组比较,黄蒲通窍胶囊组IP3表达水平明显降低(P<0.05)。结论黄蒲通窍胶囊可减少AD模型大鼠海马神经细胞的损伤,改善其学习记忆的能力,其机制可能与抑制Wnt/Ca^(2+)信号通路有关。

Objective To investigate the potential mechanism of action of Huangpu Tongqiao Capsule by observing its effect on learning and memory in a rat model of Alzheimer's disease(AD).Methods A total of 30 healthy Sprague-Dawley rats were randomly divided into blank group,model group,and Huangpu Tongqiao Capsule group,with 10 rats in each group.A rat model of AD was established by the intraperitoneal injection of D-galactose combined with injection of Aβ_(25-35) in the bilateral hippocampus.After 28 days of administration,the Morris water maze test was used to evaluate learning and memory abilities;after the samples were collected,HE staining was used to observe the damage of hippocampal neurons,Western blot was used to measure the protein expression levels of Wnt5,Frizzled,and PLCβin the hippocampus,and ELISA was used to measure the serum level of inositol triphosphate(IP3).Results The Morris water maze test showed that during the training period,the escape latency of rats in each group tended to decrease over time;during the test period,compared with the blank group,the model group had a significant increase in escape latency and a significant reduction in the percentage of distance and time in target quadrant(P<0.05),and compared with the model group,the Huangpu Tongqiao Capsule group had a significant reduction in escape latency and a significant increase in the percentage of distance and time in target quadrant(P<0.05).HE staining showed that compared with the blank group,the model group had atrophy and rupture of hippocampal neurons,a reduction in the number of cell layers,and loose arrangement with severe damage,and compared with the model group,the Huangpu Tongqiao Capsule group had a lower degree of damage of hippocampal neurons.Western blot showed that compared with the blank group,the model group had significant increases in the expression levels of Wnt5,Frizzled,and PLCβ(P<0.05),and compared with the model group,the Huangpu Tongqiao Capsule group had significant reductions in the expression levels of Wnt5,Frizzled,and PLCβ(P<0.05).ELISA showed that compared with the blank group,the model group had a significant increase in the expression level of IP3(P<0.05),and compared with the model group,the Huangpu Tongqiao Capsule group had a significant reduction in the expression level of IP3(P<0.05).Conclusion Huangpu Tongqiao Capsule can significantly alleviate the damage of hippocampal neurons and improve learning and memory abilities in a rat model of AD,possibly by inhibiting the Wnt/Ca^(2+) signaling pathway.