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24h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Aβ25-35（终浓度为10μmol／L），继续孵育24h，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。
Objective To 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 ）. Methods Cultured 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 cere-braspinal 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. Results The group of Decoction of Rehmanniae cerebrospinal fluid reduced the expression of c - jun and c - fos gene, and showed some dose - dependence. Conclusion Decoction 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.
出处
《时珍国医国药》
CAS
CSCD
北大核心
2007年第12期2875-2877,共3页
Lishizhen Medicine and Materia Medica Research
基金
国家自然科学基金资助项目(No.30472128)
关键词
地黄饮子
脑脊液
PC12细胞
即早基因
Decoction of Rehmanniae
Cerebroapinal
Pheochromocytomaderived cell line（PC12 cells）
Immediate Early gene
