黄芪注射液对脑缺血/再灌注大鼠海马组织JNK-3表达的影响

目的观察黄芪注射液对脑缺血/再灌注大鼠海马组织JNK3蛋白及其mRNA表达的影响。方法采用4VO法制备脑缺血/再灌注大鼠模型。设假手术组、模型组、黄芪注射液组和黄芪注射液溶剂对照组。除假手术组外,其余各组缺血30 min后再灌注,根据再灌注不同时间点再分为0、0.5、2、6、24、72和120 h 7个亚组。黄芪注射液组于缺血前30 min腹腔注射黄芪注射液12 g(生药)·kg-1],其中24、72、120 h组手术后每隔24 h追加给药1次,黄芪注射液溶剂对照组腹腔注射与黄芪注射液等量的无菌去离子水。分别采用HE染色观察组织形态学变化;免疫组织化学法和Western blot法检测大鼠海马组织JNK3蛋白表达的变化;RT-PCR方法检测海马组织JNK3 mRNA的表达。结果 HE染色结果显示黄芪注射液可改善脑缺血/再灌注造成的大鼠海马神经元损伤;除120 h外,模型组各时间点海马组织JNK3蛋白及mRNA表达均较假手术组增加(P<0.05);与模型组相比,黄芪注射液组除120 h外各时间点JNK3蛋白及mRNA表达均降低(P<0.05),而黄芪注射液溶剂对照组在各个时间点与模型组相比差异均无显著性(P>0.05)。结论黄芪注射液可抑制脑缺血/再灌注大鼠海马组织JNK3蛋白及其mRNA表达,从而抑制脑缺血/再灌注引起的大鼠海马神经元凋亡。

Effect of astragalus injection on expression of JNK3 in hippocampus of cerebral ischemia reperfusion rats

Aim To investigate the effect of astragalus injection on the expression of JNK3(c-Jun N terminal kinase 3) protein and JNK3 mRNA in hippocampus of cerebral ischemia reperfusion rats.Methods The rat model of cerebral ischemia reperfusion was built up by the method of four-vessel occlusion.The rats were randomly divided into 4 groups: sham operation group,model group,astragalus injection group and original astragalus injection group.Except sham operation group,the other groups after transient global cerebral ischemia(30 min),were then subdivided into 7 subsets,according to reperfusion times of 0 h,0.5 h,2 h,6 h,24 h,72 h and 120 h.The astragalus injection group was intraperitoneally injected with astragalus(12 g·kg-1) 30 min before ischemia,and the 24 h,72 h,120 h groups were given repeated administration once every 24 h after surgery.The original astragalus injection group was intraperitoneally injected with same dose of distilled water with the astragalus injection.Method of HE staining was used to observe morphological changes in hippocampus CA1 zone.Immunohistochemistry and Western blot were used to measure the expression of JNK3 protein and RT-PCR was used to measure the expression of JNK3 mRNA.Results HE staining showed that astragalus injection significantly improved pathological changes of neurons in hippocampus CA1 zone of rats.Compared with the sham operation group,the expression of JNK3 protein and mRNA in hippocampus very time points increased obviously in model group except 120 h(P<0.05).Compared with model group,the expression of JNK3 protein and mRNA in hippocampus very time points decreased obviously in astragalus injection group except 120 h(P<0.05),but there was no obvious change in original astragalus injection group(P>0.05).Conclusion Astragalus injection can inhibit the expression of JNK3 mRNA in hippocampus of cerebral ischemia and reperfusion rats,moreover,it can inhibit the expression of JNK3 protein,accordingly it inhibits hippocampal neuronal apoptosis induced by cerebral ischemia and reperfusion.