黄蒲通窍胶囊对阿尔茨海默病大鼠海马氧化应激和线粒体凋亡途径的影响

Effect of Huangpu Tongqiao Capsule on Oxidative Stress of Hippocampal Neurons and Mitochondrial Apoptotic Pathway in Rats with Alzheimer's Disease

目的探讨黄蒲通窍胶囊对阿尔茨海默病(AD)大鼠学习记忆的影响及其可能作用机制。方法SD大鼠予以腹腔注射D-半乳糖联合双侧海马注射Aβ25-35建立AD动物模型,期间给予不同剂量黄蒲通窍胶囊干预28天,同时设立假手术组和多奈哌齐阳性对照组。Morris水迷宫实验测试各组大鼠逃避潜伏期变化,HE染色观察各组大鼠海马神经元损伤情况,免疫组化法检测大鼠海马Bax表达,ELISA法检测大鼠血清中谷胱甘肽(GSH)含量,比色法检测大鼠血清中超氧化物歧化酶(SOD)活性以及丙二醛(MDA)含量,Western blot法检测大鼠海马组织细胞色素C(Cyt C)、Bcl-2、Caspase-3含量。结果与假手术组比较,模型组逃避潜伏期明显延长,海马神经元损伤严重,GSH、Bcl-2含量和SOD活性明显降低,MDA含量和Cyt C、Bax、Caspase-3表达明显升高(P<0.01);与模型组比较,黄蒲通窍胶囊组逃避潜伏期明显缩短,海马神经元损伤较轻,GSH、Bcl-2含量和SOD活性明显升高,MDA含量和Cyt C、Bax、Caspase-3表达明显降低(P<0.01,P<0.05)。结论黄蒲通窍胶囊对AD大鼠的学习记忆能力有明显改善作用,可减少AD大鼠海马神经元的损伤,其机制可能与减少氧化应激和抑制线粒体凋亡途径有关。

Objective To study the effect of Huangpu Tongqiao Capsule（ HTC） on oxidative stress and mitochondrial apoptotic pathway in rats with Alzheimer’s disease（ AD）. Methods The AD rat model was established using intraperitoneal injection of D-galactose and intracerebral injection of amyloid-β peptide25-35. Meanwhile,a sham-operation group and the Donepezil positive control group were set up. The changes of escape latency were tested using Morris water maze. The injury of hippocampal neurons was observed using HE staining. The expression of Bax was measured in hippocampal CA1 region by immunohistochemistry. The content of glutathione（ GSH） in serum was detected by ELISA. The activity of superoxide dismutase（ SOD） and the content of malonaldehyde（ MDA） in serum were detected by chromatometry,and the contents of cytochrome C（ Cyt C）,Bcl-2,and Caspase-3 in hippocampus were detected by Western blot. Results Compared with the sham-operation group,the escape latency was obviously prolonged,hippocampal neurons were severely damaged,contents of GSH and Bcl-2 as well as SOD activities obviously decreased,MDA content,and expressions of Cyt C,Bax,and Caspase-3 obviously increased inthe model group（P 〈 0. 01）. Compared with the model group,the escape latency was obviously shortened,hippocampal neurons were mildly damaged,contents of GSH and Bcl-2 as well as SOD activities obviously increased,MDA content,and expressions of Cyt C,Bax,and Caspase-3 obviously decreased in the model group（P 〈 0. 01,P 〈 0. 05）. Conclusions HTC could obviously improve the learning and memory ability of AD rats. It could reduce the damage of hippocampal neurons,which might be possibly related with reducing oxidative stress and inhibiting mitochondrial apoptotic pathway.