黄芪注射液抑制缺氧缺糖/复氧复糖大鼠海马神经元Caspase-3表达  

Effect of astragalus injection on the expression of Caspase-3 after hypoxia/hypoglycemia and reoxygenation in hippocampal neurons of rats

作　　者：闫凤霞[1] 钱涛[1] 高维娟[1] 叶冬青[1] 张雅丽[1] 侯志平[1] 

机构地区：[1]承德医学院病理生理学教研室,河北承德067000

出　　处：《解剖学报》2011年第1期14-21,共8页Acta Anatomica Sinica

基　　金：教育部新世纪优秀人才支持计划资助项目(NCET-06-0258);河北省科技领军人才创新基金资助项目(06547009D-11);河北省自然科学基金资助项目(C2006000865);河北省教育厅科学研究计划课题资助项目(2005224)

摘　　要：目的观察黄芪注射液对缺氧缺糖/复氧复糖大鼠海马神经元凋亡相关基因Caspase-3表达的影响。方法取原代培养8d的大鼠海马神经元,随机分为正常对照组、缺氧缺糖/复氧复糖组、黄芪注射液溶剂对照组和黄芪注射液组。除正常对照组外均进行缺氧缺糖0.5h再复氧复糖。各组于复氧复糖后0h、0.5h、2h、6h、24h、48h、72h和120h采用免疫组织化学染色法检测Caspase-3阳性神经元数目,采用原位杂交和RT-PCR方法检测海马神经元caspase-3 mRNA的表达,实验重复6次。结果免疫组织化学和原位杂交结果显示,与正常对照组相比,除0h和0.5h之外,缺氧缺糖/复氧复糖组各时间点海马Caspase-3蛋白、caspase-3 mRNA阳性神经元数目占神经元总数的百分率及平均吸光度值均明显增加(P<0.05),于24h达到高峰。黄芪注射液溶剂对照组以上指标的变化趋势与缺氧缺糖/复氧复糖组一致(P>0.05)。黄芪注射液组除0h和0.5h之外,各时间点以上指标比缺氧缺糖/复氧复糖组显著减少(P<0.05)。RT-PCR结果显示,除0h和0.5h外,各时间点缺氧缺糖/复氧复糖组海马神经元caspase-3 mRNA的平均吸光度值均较正常对照组明显增加(P<0.05),复氧复糖24h平均吸光度值最高;与缺氧缺糖/复氧复糖组相比,除0h和0.5h外,黄芪注射液溶剂对照组各时间点caspase-3 mRNA的平均吸光度值无明显变化(P>0.05),而黄芪注射液组在各个时间点caspase-3 mRNA的平均吸光度值明显降低(P<0.05)。结论黄芪注射液可抑制缺氧缺糖/复氧复糖大鼠海马神经元凋亡相关基因caspase-3的表达,从而抑制缺氧缺糖/复氧复糖大鼠海马神经元的凋亡。Objective To investigate the effect of astragalus injection on the expression of caspase-3 interrelated by apoptosis after hypoxia/hypoglycemia and reoxygenation in hippocampal neurons of rats. Methods Divided the hippocampal neurons cultured for eight days into four groups： normal control group （normal） , hypoxia/hypoglycemia and reoxygenation group（model） , astragalus injection solution group（solution） and astragalus injection group（astragalus）. All groups were treated with reoxygenation after deprived of oxygen and glucose for 30 minutes except normal group. Immunohistochemistry was used to measure the number of Caspase-3 positive neurons, in situ hubridization and RT-PCR were used respectively to measure the expression of caspase-3 mRNA after hypoxia/ hypoglycemia and reoxygenation 0, 0. 5,2,6,24,48,72,120 hours. The experiment was repeated 6 times. Results The results of immunohistochemistry were similar to that in in situ hybridization. It could be observed in normal control group that hippocampal neuronal nucleolus were integrated, cellular tuber was distinct and cytokinesis were dyed by yellow a lot; It could be observed in hypoxia/ hypoglycemia and reoxygenation group that hippocampal neuronal nucleolus were crimple, cellular tuber was shrinked, large numbers of cytokinesis were dyed by yellow and there were yellow granules. Compared with normal control group, the numbers, the percentage and the average absorbance （AA） of Caspase-3 protein and caspase-3 mRNA positive neuronal cells every time points increased obviously in the hypoxia/ hypoglycemia and reoxygenation group except 0 hour and 0. 5 hours（P 〈 0. 05） , and peaked at 24 hours. The change of neuronal in configuration, numbers,percentage and the average absorbance of caspase-3 mRNA positive neuronal cells in astragalus injection solution group were accorded with hypoxia/ hypoglycemia and reoxygcnation group （P 〉 0. 05 ）. Compared with hypoxia/hypoglycemia and reoxygenation group, the numbers, the percentage and th

关 键 词：海马神经元 黄芪注射液 缺氧 缺糖 复氧复糖 半胱氨酸天冬氨酸蛋白酶3 反转录一聚合酶链式 反应 大鼠 

分 类 号：R363[医药卫生—病理学]