中药更年春含药血清对β淀粉样蛋白诱导的PC12细胞凋亡的影响(英文)  

Medicated rat serum containing Gengnianchun decoction reduces apoptosis of pheochromocytoma cells insulted by amyloid beta protein

作　　者：李君[1] 李斌[1] 周文江[2] 赵凡桂[1] 李大金[1] 王文君[1] 

机构地区：[1]复旦大学妇产科医院中西医结合科,上海200011 [2]复旦大学医学院实验动物科学部,上海200032

出　　处：《中西医结合学报》2010年第5期472-479,共8页Journal of Chinese Integrative Medicine

基　　金：国家自然科学基金资助项目(No.30472259)

摘　　要：目的:研究大鼠更年春含药血清对β淀粉样蛋白25-35(amyloid beta protein 25-35,Aβ_(25-35))导致的大鼠肾嗜铬细胞瘤细胞(pheochromocytoma cell,PC12)损伤的抗凋亡作用。方法:通过对去势雌性大鼠灌服中药更年春制备含药血清,细胞计数试剂盒(cell counting kit-8,CCK-8)检测不同浓度含药血清对PC12细胞存活率的影响;不同浓度Aβ_(25-35)作用于PC12细胞,建立阿尔茨海默病细胞模型,CCK-8法检测含药血清对损伤细胞的保护作用,流式细胞术检测PC12细胞在Aβ_(25-35)和含药血清共同培养下的细胞凋亡率;蛋白印迹法检测各组细胞Bcl-2、Bax和半胱氨酸天冬氨酸特异性蛋白酶3(caspase-3)的蛋白表达情况。结果:与空白血清相比,20%含药血清培养PC12细胞24 h或48 h后可促进其增殖(P<0.05)。Aβ_(25-35)对PC12细胞有细胞毒性,并呈剂量依赖性地降低细胞存活率(P<0.05),导致细胞凋亡。CCK-8法和流式细胞术检测均显示,20%更年春含药血清对Aβ_(25-35)诱导的PC12细胞损伤有保护作用(P<0.05),其作用效果类似于阳性对照药神经生长因子。蛋白印迹法检测显示,与模型组比较,含药血清组Bax和Bcl-2蛋白表达的比值和caspase-3蛋白表达均降低。结论:更年春含药血清可提高Aβ_(25-35)损伤的PC12细胞的存活率,并减少PC12细胞凋亡。其机制可能与调控Bcl-2家族蛋白的表达而实现抗凋亡作用有关。Objective：To investigate the effects of medicated rat serum containing Gengnianchun（GNC） decoction and its protection to pheochromocytoma cells（PC12 cells） from amyloid beta（Aβ）25-35-insulted apoptosis and to find the possible mechanism. Methods：Medicated rat serum was prepared by administering ovariectomized Sprague-Dawley（SD） rats with GNC decoction.The effects of medicated rat serum on viability of PC12 cells were evaluated by cell counting kit-8（CCK-8） assay.The PC12 cells were cultured with different doses of Aβ25-35 to induce a model of Alzheimer＇s disease in vitro.Then,the protective effects of medicated rat serum on Aβ25-35-insulted PC12 cells were evaluated by using CCK-8 assay to detect the cell viability,using AnnexinⅤ-fluorescein isothiocyanate（FITC）/propidium iodide（PI） flow cytometry to detect cell apoptosis rate and using Western blotting assay to analyze the expressions of Bcl-2,Bax and active caspase-3 proteins. Results：PC12 cells cultured with 20%medicated rat serum containing GNC decoction for 24 h or 48 h had higher viability than those cultured with normal culture medium（P〈0.05）.After 24- or 48-hour treatment of different concentrations of Aβ25-35,cell viabilities were all decreased as compared with normal medium（P〈0.05）.Cells underwent apoptosis,which showed the neurotoxicity of Aβ25-35.The cell apoptosis induced by Aβ25-35 was significantly decreased in PC12 cells which were pretreated with 20%medicated rat serum or nerve growth factor（NGF） according to CCK-8 assay and Annexin V-FITC/PI flow cytometry（P〈0.05）. The ratio of Bax expression to Bcl-2 expression and the expression of active caspase-3 were decreased in the cells treated with medicated serum or NGF as compared with the cells cultured with Aβ25-35 only. Conclusion：The GNC-medicated rat serum at concentration of 20%can promote viability of Aβ25-35-insulted PC12 cells and decrease the cell apoptosis by regulating the expressions of Bcl-2,Bax and active caspase 3.

关 键 词：阿尔茨海默病 凋亡 含药血清 β淀粉蛋白 BCL-2 

分 类 号：R285.5[医药卫生—中药学]