地黄饮子对海马神经元AD模型细胞凋亡的调控作用

Effect of Decoction of Rehmanniae on the Expression of Apoptosis of Hippocampal Neurons Model of Alzeimer Disease

目的 :探讨古方地黄饮子含药脑脊液对 β 淀粉样蛋白 (Aβ2 5～ 35)所致海马神经细胞AD(Alzheimer’sDisease)模型凋亡基因bcl 2、bax和caspase 3表达的影响。方法 :运用中药脑脊液药理学的方法把体外培养成熟的大鼠海马神经细胞分为空白组、模型组、VitE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。然后除空白组加入等量培养液外 ,其它各组加入经老化处理的Aβ2 5～ 35(终浓度为 1 0 μmol/L)共同孵育 2 4h ,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bcl 2、bax和caspase 3基因的表达。结果 :地黄饮子脑脊液能剂量依赖性地增强受损海马神经元bcl 2基因的表达 ,降低bax和caspase 3基因的表达。说明对Aβ2 5～ 35所致离体海马神经细胞凋亡有抑制作用 ,其作用机理可能与地黄饮子升高bcl 2 /bax的比值 ,抑制线粒体释放细胞色素C ,控制caspase

Objective:To investigate effects on the expression of apoptosis gene bcl 2、bax and caspase 3 of Decoction of Rehmanniae on cultured hippocampal neurons injury induced by β amyloid 25～35 protein(A β 25～35 ).Methods:Cultured hippocampal neurons were divided into 6 groups by using the cerebrospinal fluid with Traditional Chinese medicine normal group、model group、vitamin E group、Low dose of TCM group、moderate dose of TCM group and high dose of TCM group.Six group were respectively added to normal cultured fluid、normal cerebrospinal 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 A β 25～35 dealed with aging(the final concentration is 10μmol/L).and then six groups were hatched 24 hours together. At last neurous were digested by trypsin,centrifuged,collected and then distilled the total RNA.The expression of bcl 2、bax and caspase 3 by using the methods of RT PCR and agarose gelelectrophoresis were measured. Results:The group of Decoction of Rehmanniae cerebrospinal fluid can dose dependedly enhance the expression of bcl 2 gene,reduce the expression of bax and caspase 3 gene injured hippocampal neurons.It showed that Decoction of Rehmanniae cerebrospinal fluid had the inhibitory effects on apoptosis of cultured Hippocampal neurons injury induced by A β 25～25 . The mechanism may associate that Decoction of Rehmanniae enchanced the ratio of bcl 2/bax.restrained mitochondrial from releasing cytochrome c,controlled the activation of caspase 3