黄芪注射液抑制缺氧缺糖/复氧复糖大鼠海马神经元JNK3 mRNA的表达

Astragalus injection inhibits the expression of JNK3 mRNA 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基因c-Jun氨基末端激酶3(JNK3)mRNA表达的影响。方法:取原代培养8 d的大鼠海马神经元,随机分为4组:正常对照组、黄芪注射液原液对照组、缺氧缺糖/复氧复糖组和黄芪注射液组。其中缺氧缺糖/复氧复糖组、黄芪注射液组及黄芪注射液原液对照组进行缺糖缺氧0.5 h再复氧复糖,并于复氧复糖后0 h、0.5 h、2 h、6 h、24 h、72 h和120 h采用原位杂交和RT-PCR方法检测海马神经元JNK3 mRNA的表达。结果:正常对照组大鼠海马神经元核膜完整,细胞突起明显,有少许细胞胞浆黄染;缺氧缺糖/复氧复糖组大鼠海马神经元胞核体积增大、突起回缩,大量细胞胞浆黄染,胞核内有黄色颗粒,海马JNK3 mRNA阳性神经元数目在各个时点均比正常对照组明显增多(P<0.05);黄芪注射液原液对照组神经元形态变化和海马JNK3 mRNA阳性神经元数目均与缺氧缺糖/复氧复糖组相一致(P>0.05);黄芪注射液组大鼠海马神经元有轻微核皱缩,可见部分神经元胞浆黄染,除120 h时点之外,海马JNK mRNA阳性神经元数目在各个时点均比缺氧缺糖/复氧复糖组明显减少(P<0.05)。缺氧缺糖/复氧复糖组在0 h、0.5 h、2 h、6 h、24 h、72 h和120 h海马神经元JNK3 mRNA平均灰度值均较正常对照组明显增加(P<0.05);与缺氧缺糖/复氧复糖组相比,黄芪注射液原液对照组各时点JNK3 mRNA的平均灰度值无明显变化(P>0.05),而黄芪注射液组除120 h之外在各个时点JNK3mRNA的平均灰度值均明显降低(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JNK3 mRNA表达,从而抑制缺氧缺糖/复氧复糖大鼠海马神经元的凋亡。

AIM： To investigate the effect of astragalus injection on the expression of c - jun N terminal kinase （JNK3） mRNA interrelated with apoptosis after hypoxia/hypoglycemia and reoxygenation in hippocampal neurons of rats. METHODS： The hippocampal neurons cultured for eight days were divided into 4 groups： normal control group, original astragalus injection group, hypoxia/hypoglycemia and reoxygenation group, astragalus injection group. Hypoxia/hypoglycemia and reoxygenation group, astragalus injection group and original astragalus injection group were treated with hypoglycemia and reoxygenation after deprived of oxygen and glucose for 30 min. Methods of in situ hybridization and RT - PCR were used respectively to measure the expression of JNK3 mRNA after hypoxia/hypoglycemia and reoxygenation 0 h, 0. 5 h, 2 h, 6 h, 24 h, 72 h and 120 h. RESULTS： In normal control group the volume of hippocampal neuronal nucleolus was accretion, cellular tuber was distinct and cytokinesis was dyed by yellow a lot. In hypoxia/hypoglycemia and reoxygenation group the hippocampal neuronal nucleolus was crimpte, cellular tuber was shrinked, large number of cytokinesis was dyed by yellow and yellow granule was observed. Compared with normal control group, the numbers of JNK3 mRNA positive neuronal cells at each time point increased obviously in the hypoxia/hypoglycemia and reoxygenation group （P 〈0. 05 ）. The change of neuronal configuration and the numbers of JNK3 mRNA positive neuronal cells in original astragalus injection group accorded with hypoxia/hypoglycemia and reoxygenation group （ P 〉 0.05 ）. In astragalus injection group the hippocampal neuronal nucleolus was crimple slightly and segmental cytokinesis was dyed by yellow. Compared to hypoxia/ hypoglycemia and reoxygenation group, the numbers of JNK3 mRNA positive neuronal cells at each time point were less obviously in the astragalus injection group besides 120 h （P 〈 0. 05）. Compared to normal control group, the mean optic density of expression of JNK3 mRNA in hippocampal neurons of rats at each time point increased obviously in hypoxia/hypoglycemia and reoxygenation group （ P 〈 0. 05）. Compared to hypoxia/hypoglycemia and reoxygenation group, the mean optic density of JNK3 mRNA expression at each time point in original astragalus injection group had no obvious change （ P 〉 0. 05）, however the mean optic density of JNK3 mRNA expression in hippocampal neurons of rats at each time point decreased obviously in the astragalus injection group besides 120 h （P 〈 0. 05 ）. CONCLUSION： Astragalus injection inhibits the expression of JNK3 mRNA after hypoxia/hypoglycemia and reoxygenation, accordingly inhibits hippocampal neuronal apoptosis.