摘要
背景:淀粉样β蛋白是老年斑的核心成分,其毒性片段淀粉样β蛋白25-35近年广泛用于实验研究中。已往研究表明自制中药复智散可促进培养神经细胞存活,可能具备治疗阿尔茨海默病的效用。目的:研究复智散对抗淀粉样β蛋白25-35对培养神经细胞的毒性作用及其发挥这一作用的可能途径。设计:以细胞为研究对象,重复测量设计。单位:一所大学医院的神经内科和一所大学医院的脑老化重点实验室。材料:实验于2002-06/2003-04在宣武医院北京脑老化重点实验室完成。人多巴胺能神经母细胞瘤株SH-SY5Y细胞。淀粉样β蛋白25-35片段。中药复智散文火煎制,留取上清冷冻抽干配成浓度为0.5g/mL的贮存液备用。抗体:cAMP应答元件结合蛋白、B淋巴细胞白血病-2基因产物中的Bcl-2、细胞色素C由宣武医院北京脑老化研究实验室制备。方法:用不同剂量的淀粉样β蛋白25-35单独或与复智散共同孵育SH-SY5Y细胞,与空白对照组相比,测定在不同孵育条件下培养神经细胞的噻唑蓝代谢率。利用Western-blot法测定复智散单独孵育、淀粉样β蛋白25-35单独孵育及两者共同孵育条件下与空白对照相比蛋白质表达的变化。主要观察指标:各实验组与空白对照组相比的噻唑蓝代谢率。与神经细胞存活及死亡相关蛋白cAMP应答元件结合蛋白。
BACKGROUND:Amyloid β(Aβ) protein is the core of senile plaque. Being the toxic segment of Aβ,Aβ25 35 has been extensively applied in the experiments of recent years.The research in the past has verified that the self prepared Chinese herb,fuzhisan can promote the survival of the cultured neural cells and probably acts on the treatment of Alzheimer disease(AD). OBJECTIVE:To study the resistance of fuzhisan to Aβ25 35 toxicity to cultured neural cells and the probable approaches. DESIGN:Repeated measurement based on the cells. SETTING: Department of neurology of a university hospital and key experimental room in brain aging in a university hospital. MATERIALS:The experiment was performed in Beijing Key Experimental Room in Brain Aging of Xuanwu Hospital from June 2002 to April 2003.Dopaminergic SH SY5Y cell of neuroblastoma and Aβ25 35 were employed.Chinese herb,fuzhisan was decocted with mild fire and its upper clear solution was collected and prepared into storage solution at the concentration of 0.5 g/mL.Antibody: Beijing Key Experimental Room in Brain Aging of Xuanwu Hospital prepared cAMP responsive element binding protein(CREB),Bcl 2 in B lymphatic leukaemia 2 genetic product and cytochrome C(CytC). METHODS:SH SY5Y cell was incubated with Aβ25 35 of various doses alone or in combination with fuzhisan and was compared with blank control.MTT metabolic rate of cultured neural cells were determined under different incubation conditions.Western blot method was used to measure the protein expression changes in incubation with fuzhisan alone,incubation with Aβ25 35 alone and the combination incubation,compared with the blank control. MAIN OUTCOME MEASURES:It was to study MTT metabolic rate in the comparison between each experimental group and the blank control and expressions of CREB,Bcl 2 and CytC relevant to survival/death of neural cells. RESULTS:Survival rate of SH SY5Y cell was increased by 11.4%in incubation with fuzhisan alone.It was remarkably improved in incubation combining fuzhisan with Aβ25 35 as compared with Aβ25 35 alone.The expressions of CREB and Bcl 2 in Aβ25 35 group were decreased and were increased in fuzhisan group.CytC expression in cytoplasm was increased in Aβ25 35 group and was declined with fuzhisan incubation. CONCLUSION:Fuzhisan promotes the survival of cultured neural cells and its protection is still existed under Aβ25 35 injury.Fuzhisan brings such effects into play probably by the protein expressions relevant to survival/death of cells.
出处
《中国临床康复》
CSCD
北大核心
2005年第13期248-249,F003,共3页
Chinese Journal of Clinical Rehabilitation
基金
黑龙江省九五攻关课题(Z99C20-24)~~
