黄芪注射液抑制缺氧缺糖/复氧复糖大鼠海马神经元JNK3的表达

Astragalus injection inhibits the expression of JNK3 after hypoxia/hypoglycemia and reoxygenation in hippocampal neurons of rats

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蛋白c-Jun氨基末端激酶3(c-Jun N ter-minal kinase 3,JNK3)及其mRNA表达的影响,探讨黄芪注射液防治海马神经元凋亡可能的作用机制。方法取原代培养8d的大鼠海马神经元,随机分为4组:正常对照组、缺氧缺糖/复氧复糖组、黄芪注射液组和黄芪溶剂对照组。其中缺氧缺糖/复氧复糖组、黄芪注射液组和黄芪溶剂对照组进行缺氧缺糖0.5h再复氧复糖,并于复氧复糖后0、0.5、2、6、24、72和120h采用Western blot、ELISA和RT-PCR方法检测海马神经元JNK3蛋白及其mRNA的表达。结果缺氧缺糖/复氧复糖组在0、0.5、2、6、24和72h海马神经元JNK3蛋白条带平均光密度值及蛋白活性均较正常对照组增加(P<0.05),而复氧复糖120h组较正常对照组降低(P<0.05);与缺氧缺糖/复氧复糖组相比,除120h之外,黄芪注射液组在各个时间点JNK3蛋白条带的平均光密度值及蛋白活性均降低(P<0.05);而黄芪溶剂对照组与缺氧缺糖/复氧复糖组相比则差异无显著性(P>0.05)。缺氧缺糖/复氧复糖组在0、0.5、2、6、24、72和120h海马神经元JNK3mRNA平均光密度值均较正常对照组增加(P<0.05);与缺氧缺糖/复氧复糖组相比,除120h之外,黄芪注射液组在各个时间点JNK3mRNA的平均光密度值均降低(P<0.05);而黄芪溶剂对照组与缺氧缺糖/复氧复糖组相比则差异无显著性(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JNK3mRNA表达,从而抑制JNK3蛋白表达,降低了JNK3蛋白活性,从而抑制缺氧缺糖/复氧复糖引起大鼠海马神经元凋亡的作用。

Aim To investigate the effect of astragalus injection on the expression of JNK3（c-jun N terminal kinase）protein and JNK3 mRNA interrelated by apoptosis after hypoxia/hypoglycemia and reoxygenation in hippocampal neurons of rats. Methods The hipp-ocampal neurons cultured for eight days were divided into four groups：normal control group,hypoxia/hypo-glycemia and reoxygenation group,astragalus injection group and astragalus solution group. Hypoxia/hypogly-cemia and reoxygenation group,astragalus injection group and astragalus solution group were treated with hypoglycemia and reoxygenation after being deprived of oxygen and glucose for 30 minutes. Methods of Western blot,ELISA and RT-PCR were used respectively to measure the expression of JNK3 mRNA after hypoxia/hypoglycemia and reoxygenation 0,0. 5,2,6,24,72, 120 h. Results Compared with normal control group, the mean optic density（ MOD） of expression of JNK3 protein and activation of JNK3 protein in hippocampal neurons of rats every time points increased obviously in hypoxia/hypoglycemia and reoxygenation group except 120 h（P〈0.05）;compared with hypoxia/hypoglycemia and reoxygenation group,MOD of expression of JNK3 mRNA and activation of JNK3 protein in hipp-ocampal neurons of rats every time points decreased obviously except 120 h in astragalus injection group （P〈0. 05）;compared with hypoxia/hypoglycemia and reox-ygenation group,there was no difference in astragalus solution group. Compared with normal control group, MOD of expression of JNK3 mRNA in hippocampal neurons of rats every time points increased obviously in hypoxia/hypoglycemia and reoxygenation group （P〈0.05）; compared with hypoxia/ hypoglycemia and reoxygenation group,MOD of expression of JNK3 mRNA in hippocampal neurons of rats every time points decreased obviously in astragalus injection group except 120 h（P〈0.05）; compared with hypoxia/hypoglycemia and reoxygenation group,there was no difference in astragalus solution group. Conclusion Astragalus injection can inhibit the expression of JNK3 mRNA after hypoxia/hypoglycemia and reoxygenation,moreover,it can inhibit the expression of JNK3 protein and decrease the activation of JNK3 protein,accordingly it inhibits hippocampal neuronal apoptosis.