地黄饮子对海马神经元损伤即早基因c-jun和c-fos表达的影响

Effect of Decoction of Rehmanniae on Immediate Early Gene c-jun and c-fos of Hippocampal Neurons Model injury

目的 :探讨古方地黄饮子对 β淀粉蛋白 (Aβ2 5～ 3 5)所致离体海马神经元损伤时即早基因c jun和c fos表达的调控作用。方法 :把体外培养的大鼠海马神经细胞分为正常组、模型组、VitE脑脊液对照组、地黄饮子脑脊液低、中、高剂量组 ,6组分别加入正常培养液、正常脑脊液、VitE脑脊液、中药脑脊液低、中、高剂量 ,继续孵育 2h。然后除正常组加入等量培养液外 ,其他各组加入经老化处理的Aβ2 5～ 3 5(终浓度为 10 μmol L)共同孵育 1h ,胰蛋白酶消化 ,离心收集细胞 ,提取总RNA。应用RT PCR和琼脂糖凝胶电泳方法测定c jun和c fos基因的表达。结果 :地黄饮子脑脊液能剂量依赖性地降低受损海马神经元c jun和c fos基因的表达。结论 :地黄饮子脑脊液能有效降低海马神经元受到神经毒损伤时的应激反应 ,降低c jun、c

Objective:To investigate repressive effects on the expression of IEG c jun and c fos of Decoction of Rehmanniae on cultured hippocampal neurons injury induced by β amyloid 25～35 protein(Aβ 25～35 ).Methods :Cultured hippocampal neurons were divided into 6 groups:normal group,model group,vitamin E group、low dose of TCM group,moderate dose of TCM group and high dose of TCM group.Six groups were respectively added to normal cultured fluid,normal cerebrospmal fluid、vitE cerebrospinal fluid,low dose of TCM cerebrospinal fluid,moderate and high dose of cerebrospinal fluid,hatched 2 hours.The normal group was added to the same dose of cultured fluid and other groups was added to Aβ 25～35 dealed with aging(the final concentration is 10 μmol/l),and then six groups were hatched 1 hour together.At last neurons were digested by trypsin,centrifuged,got together and then distilled the total RNA.It can be measured the expression of c jun and c fos by using the methods of RT PCR and agarose gel electrophoresis.Results The group of Decoction of Rehmanniae cerebrospinal fluid can dose dependedly reduce the expression of c jun and c fos gene injured hippocampal neurons.Conclusion Decoction of Rehmanniae cerebrospinal fluid can effectively reduce the stress response of hippocampal neurons injured by neurotoxin and reduce the neuron injury induced by c jun and c los gene.