参芎化瘀胶囊对全脑缺血大鼠海马CA1区GAP-43 和P75NTR表达的影响

Influence on the Expression of GAP-43 and P75NTR after Global Cerebral Ischemia-reperfusion in Hippocampus by Shenxiong Huayu Capsules

摘要:

目的: 探讨参芎化瘀胶囊对全脑缺血大鼠海马CA1区生长相关蛋白-43(GAP-43)和神经营养因子低亲合力受体(P75NTR)表达的影响。方法: 清洁级SD雄性大鼠72只,分正常对照组、全脑缺血模型组和参芎化瘀胶囊+全脑缺血组(灌胃剂量:480 mg·kg^-1·d^-1,每日1次,早上8:00给药),每组24只,取3,7,14 d 3个时间点。制备全脑缺血模型。水迷宫评价学习记忆能力,HE染色观察组织病理学的变化,免疫组化法观察GAP-43表达,蛋白印迹分析检测GAP-43和P75NTR蛋白含量。结果: ①行为学:和正常对照组比较,模型组大鼠各个时间点逃避潜伏期时间明显延长(P<0.05),参芎化瘀胶囊组大鼠逃避潜伏期时间较模型组缩短(P<0.05);②HE 染色显示:与正常组比较,模型组大鼠海马CA1区神经元3~14 d进行性坏死,存活神经元减少,参芎化瘀胶囊组各个时间点正常神经元计数和模型组比较有差异(P<0.05);③GAP-43免疫组化和蛋白印迹 结果:和正常组比较,模型组GAP-43的表达3 d升高,7 d最明显,14 d下降;和模型组比较,参芎化瘀胶囊组3,7,14 d持续性升高,各个时间比较均有差异(P<0.05);④P75NTR蛋白印迹结果分析:和正常组比较,模型组的表达3 d升高,7 d最明显,14 d下降;和模型组比较,参芎化瘀胶囊组3,7,14 d持续性升高,所有时间点比较均有差异(P<0.05)。结论: 参芎化瘀胶囊增强全脑缺血大鼠海马CA1区GAP-43和P75NTR的表达。

Abstract:

Objective: To study the effect of Shenxiong Huayu capsules on growth associated protein 43(GAP-43) and nerve growth factor receptor(P75NTR) expression after global cerebral ischemia-reperfusion in hippocampus. Method: Adult male SD rats (n=72) were randomly divided into 3 groups:sham group, model group (I/R) and Shenxiong Huayu capsule group,24 rats in each group and was divided into 3 subgroups:3, 7, 14 day. The model of global ischemia-reperfusion was formed by improved Pulsinelli's 4-vessel occlusion method. The behavior abilities of learning and memory were observed by using Morris water maze. Morphological changes were observed by HE staining in 3, 7, 14 day after injury. Immunohistochemistry method was applied to observe the expression of GAP-43.Western-blotting method was used to detect the protein content of GAP-43 and P75NTR. Result: Compared with normal control group, the learning and memory abilities of rats in model group were significantly declined (P<0.05). The ability of learning and memory of the rats were improved after treated by Shenxiong Huayu capsules (P<0.05).The time of escaping latency was shortened than model group. Compared with normal control group, model group's neuronal morphous was damaged. Survival cells were declined. Compared with model group, Shenxiong Huayu capsules group had more normal cells in hippocampus (P<0.05). In model group, the expression of GAP-43 was increased from the third day (P<0.05) and declined on the 14th day;compared with model group, the expression of GAP-43 was increased significantly in Shenxiong Huayu capsules group on every time point (P<0.05).In model group, the protein contain of P75NTR was increased from the third day(P<0.05)and declined on the 14th day. Compared with model group, the protein contain of P75NTR was increased significantly in Shenxiong Huayu capsules group on every time point(P<0.05). Conclusion: Shenxiong Huayu capsules have heightened the expression of GAP-43 and P75NTR on global cerebral ischemia-reperfusion in hippocampus.