不同时间点给予参芎化瘀胶囊对脑缺血/再灌注损伤大鼠的神经保护作用

摘要

目的：探讨脑缺血/再灌注损伤后不同时间点给予参芎化瘀胶囊（每天480mg/100g）对大鼠海马区神经元的保护作用及机制。方法 SD大鼠60只随机分为假手术组、模型组和参芎化瘀胶囊治疗组，线栓法制备左侧大脑中动脉栓塞/再灌注模型，每组根据术后干预时间点再分为术后6、12、24及48 h共4个亚组。参芎化瘀胶囊治疗组于动物缺血再灌注后不同时间点给予参芎化瘀胶囊治疗，于治疗14d后取脑组织采用HE染色观察海马区神经元形态的变化及存活神经元数量，免疫组化和Werston blot法检测海马区P75NTR和Notch1蛋白的表达。结果造模后不同时间点给予参芎化瘀胶囊治疗的各亚组海马区存活神经细胞数量较模型组多，P75NTR和Notch1蛋白表达水平增高（P<0.05）；干预时机效果的比较显示，造模后12h给予参芎化瘀胶囊组存活神经元数量、P75NTR和Notch1蛋白表达水平最高，差异有统计学意义（P<0.05）。结论参芎化瘀胶囊具有保护神经元及促进脑缺血再灌注大鼠海马区P75NTR、Notch1蛋白表达的作用，在脑缺血再灌注损伤12h内给予效果最佳。%Objective To investigate effect of Shenqiong Huayu capsule in different time windows on neuroprotective and expression of nerve P75NTR and Notch1 in hippocampal after cerebral ischemia reperfusion of rats. Methods Adult male SD rats(n=60) were randomly divided into 3 groups:sham operated group,ischemia reperfusion and Shenqiong Huayu capsule group. Every group was further divided into 6h,12h,24h and 48h subgroups. The cerebral infarction model was established by left middle cerebral artery occlusion. The rats in the Shenqiong Huayu capsule group were treated with Shenqiong Huayu at different time points after the operation and the animals in the other two groups were treated with sham-Shenqiong Huayu at the same time point s as that in Shenqiong Huayu group. Morphological changes were observed by HE staining after injury. Immunohistochemistry method Werston-blot method were applied to observe the expression of P75NTR and Notch1 at day 14 after therapy. Results There was a significant difference in the expression of P75NTR and Notch1 and the survival rate of nerve cells between Shenqiong Huayu group and the other two groups at the same time points (P<0.05). In Shenqiong Huayu group,the level of P75NTR and Notch1 was the highest in 12h subgroup (P<0.05). Conclusion Shenqiong Huayu capsules had protective effect on cerebral IR in the hippocampal and promote their hippocampal neurogenesis of SD rat,which might be associated with increased expression of P75NTR and Notch1. Shenqiong Huayu capsule treatment could up-regulate the expression P75NTR and Notch1.The effects reach the peak when Shenqiong Huayu treatment was administered in the 12h time window after reperfusion.