地黄饮子含药脑脊液对Aβ25-35诱导PC12细胞损伤即早基因c-fos及c.docVIP

地黄饮子含药脑脊液对Aβ25-35诱导PC12细胞损伤即早基因c-fos及c 　 作者：谢宁 孟菲 姚辛敏 周妍妍 何秀丽 【摘要】 　　目的探讨古方地黄饮子对β淀粉酶Aβ25～35所致PC12细胞损伤时即早基因c-fos和c-jun表达的调控作用。方法把离体培养的进入对数生长期的PC12细胞分为空白组、模型组、VitE脑脊液对照组、地黄饮子脑脊液低、中、高剂量组，待24 h细胞贴壁后吸去培养液,6组分别加入正常培养液、正常脑脊液、VitE脑脊液、地黄饮子脑脊液低、中、高剂量，加培养液补至等量，37℃孵育2 h。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Aβ25～35（终浓度为10 μmol/L），继续孵育24 h，然后离心收集细胞，提取总RNA。应用RT-PCR二步法和琼脂糖凝胶电泳方法测定c-fos和c-jun基因的表达。结果地黄饮子能下调c-fos和c-jun基因的表达，并呈一定的剂量依赖性。结论地黄饮子脑脊液能明显降低PC12细胞受Aβ25～35刺激时即早基因c-fos和c-jun的过度反应，说明地黄饮子能提高PC12细胞对外界不良刺激的应激性。 【关键词】 地黄饮子 脑脊液 PC12细胞 即早基因 　　Abstract:ObjectiveTo investigate the regulative effects on the expression of IEG c-jun and c-fos of Decoction of rehmanniae on PC12 injury induced by b-amyloid25～35 protein(Aβ25～35).MethodsCultured cells of PC12 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;nomal cerebrospinal fluid,vitE cerebrospinal fluid,low dose of TCM cerebraspinal fluid, moderate and high dose of TCM cerebrospinal fluid in log growth phase,then added cultured fluid to equal,37℃ hatched 2 hours.The normal group was added to the same dose of cultured fluid and other groups were added to Aβ25～35 dealed with aging (the final concentration is 10 μmol/L),and then six groups were hatched 24 hours together.At last they were centrifuged to extract the total RNA. The expression of c-jun and c-fos measured by the methods of RT-PCR and agarose gel electrophoresis.ResultsThe group of Decoction of Rehmanniae cerebrospinal fluid reduced the expression of c-jun and c-fos gene, and showed some dose-dependence.ConclusionDecoction of Rehmanniae cerebrospinal fluid can effectively reduce the high response of c-jun and c-fos gene in PC12 cells injured by Aβ25～35.It shows that Decoction of Rehmanniae cerebrospinal fluid can increase PC12 cells’ irritability for external ill innervation. 　　Key words:Decoction of Rehmanniae; Cerebroapinal; Pheochromoc