复智散对淀粉样β蛋白25-35神经细胞毒性效应的影响及其机制
복지산대정분양β단백25-35신경세포독성효응적영향급기궤제
Effect and mechanism of fuzhisan on toxic reaction of amyloid beta 25-35 to neural cells
  • 万方数据
    中国临床康复 중국림상강복
    CHINESE JOURNAL OF CLINICAL REHABILITATION
    2005年 13期 248-249 ,共2页

    温世荣%王德生%张景艳%盛树力

    온세영%왕덕생%장경염%성수력

    阿尔茨海默病%中药%淀粉样β蛋白 阿爾茨海默病%中藥%澱粉樣β蛋白
    아이자해묵병%중약%정분양β단백
    .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0.5 g/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,Bcl-2及细胞色素C的表达水平.结果:复智散单独孵育可使SH-SY5Y细胞存活率增加11.4%,与淀粉样β蛋白25-35共同孵育时培养神经细胞存活率较淀粉样β蛋白25-35单独孵育明显提高.淀粉样β蛋白25-35损伤组cAMP应答元件结合蛋白和Bcl-2表达减少,复智散组这两种蛋白质表达增加.淀粉样β蛋白25-35损伤组胞浆内细胞色素C表达增加,复智散孵育下该蛋白质表达水平下降.结论:复智散有促进培养神经细胞存活作用,在淀粉样β蛋白25-35损伤条件下该种保护作用依旧存在.复智散可能通过影响细胞存活/死亡相关蛋白质的表达发挥这一效应.
    .배경:정분양β단백시노년반적핵심성분,기독성편단정분양β단백25-35근년엄범용우실험연구중.이왕연구표명자제중약복지산가촉진배양신경세포존활,가능구비치료아이자해묵병적효용.목적:연구복지산대항정분양β단백25-35대배양신경세포적독성작용급기발휘저일작용적가능도경.설계:이세포위연구대상,중복측량설계.단위:일소대학의원적신경내과화일소대학의원적뇌노화중점실험실.재료:실험우2002-06/2003-04재선무의원북경뇌노화중점실험실완성.인다파알능신경모세포류주SH-SY5Y세포.정분양β단백25-35편단.중약복지산문화전제,류취상청냉동추간배성농도위0.5 g/mL적저존액비용.항체:cAMP응답원건결합단백、B림파세포백혈병-2기인산물중적Bcl-2、세포색소C유선무의원북경뇌노화연구실험실제비.방법:용불동제량적정분양β단백25-35단독혹여복지산공동부육SH-SY5Y세포,여공백대조조상비,측정재불동부육조건하배양신경세포적새서람대사솔.이용Western-blot법측정복지산단독부육、정분양β단백25-35단독부육급량자공동부육조건하여공백대조상비단백질표체적변화.주요관찰지표:각실험조여공백대조조상비적새서람대사솔.여신경세포존활급사망상관단백cAMP응답원건결합단백,Bcl-2급세포색소C적표체수평.결과:복지산단독부육가사SH-SY5Y세포존활솔증가11.4%,여정분양β단백25-35공동부육시배양신경세포존활솔교정분양β단백25-35단독부육명현제고.정분양β단백25-35손상조cAMP응답원건결합단백화Bcl-2표체감소,복지산조저량충단백질표체증가.정분양β단백25-35손상조포장내세포색소C표체증가,복지산부육하해단백질표체수평하강.결론:복지산유촉진배양신경세포존활작용,재정분양β단백25-35손상조건하해충보호작용의구존재.복지산가능통과영향세포존활/사망상관단백질적표체발휘저일효응.
    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. MTF 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 fuzhisanalone, 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 fu zhisan 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.
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