淀粉样β蛋白25-35诱导兔主动脉平滑肌细胞凋亡的研究  

Amyloid-beta protein 25-35 induced apoptosis in rabbit aortic smooth muscle cells:an experimental study

作　　者：周小英[1] 张微微[1] 

机构地区：[1]北京军区总医院神经内科

出　　处：《中国脑血管病杂志》2007年第2期68-71,共4页Chinese Journal of Cerebrovascular Diseases

摘　　要：目的探讨淀粉样β蛋白25-35(Aβ25-35)体外诱导主动脉血管平滑肌细胞(VSMC)凋亡作用。方法取正常兔胸主动脉VSMC株,将其分为实验组和对照组。实验组为Aβ25-35与VSMC共培养,按Aβ25-35浓度再分为10、20、50及100μmol/L组;对照组为等渗盐水与VSMC共培养。每组分别培养1、3及10d。采用AnnexinV-FITC、碘化丙啶染色法定性检测细胞凋亡情况;借助流式细胞仪定量观察细胞凋亡情况。结果Aβ25-3510μmol/L组孵育3d时,VSMC凋亡率为(5.3±1.4)%,20μmol/L组为(25.9±5.8)%;Aβ25-35100μmol/L组孵育10d时,凋亡率为(94.0±14.4)%,3组比较差异均有统计学意义(P<0.05),凋亡率随Aβ25-35浓度、孵育时间的增加而增高。Aβ25-3520μmol/L组孵育1d后,即可观察到处于早期凋亡的细胞呈现绿色荧光标记;Aβ25-35100μmol/L组孵育10d,可观察到呈现绿色、红色两种荧光标记,为处于凋亡晚期或临近坏死的细胞。结论Aβ25-35可通过凋亡途径发挥对VSMC的毒性作用。Objective To investigate the effect of amyloid-beta protein 25-35 （ Aβ25-35 ） induced apoptosis in cultured vascular smooth muscle cells （VSMCs） in vitro. Methods The normal VSMCs of rabbit thoracic aorta were selected, and they were divided into experimental and control groups. The experimental group was incubated with Aβ25-35 and VSMCs, and they were redivided into 10, 20, 50, and 100 μmol/Lgroups according to their Aβ25-35 concentrations; the control group was incubated with isotonic saline solution and VSMCs. Each group was incubated for 1,3, and 10 d, respectively. Apoptosis was detected with annexin V-fluorescein isothiocyanate （V-FITC） and propidium iodide （PI） staining methods. Apoptosis was observed quantitatively with flow cytometer. Results The apoptosis rates of VSMCs were 5.3 ± 1.4% and 25.9 ± 5.8% incubated for 3 d in the 10 and 20 μmol/L Aβ25-35 groups. The apoptosis rate was 94. 0 ± 14. 4% incubated for 10 d in the 100 μmol/L Aβ25-35 group, and these differences were significant （P 〈0. 05）. The apoptosis rates increased with the Aβ25-35 concentrations and incubation time. The early apoptotic cells presented green fluorescent label which could be observed in the 20μmol/L Aβ,25-35 group incubated for only 1 d ; both green and red fluorescent labels could be observed in the 100 μmol/L Aβ25-35 group incubated for 10 d, and they were in the late apoptotic stage or the dying cells. Conclusion Aβ25-35 may exert cytotoxic effects on VSMC through apoptotic pathway.

关 键 词：淀粉样Β蛋白 胸部动脉 肌细胞 平滑肌 凋亡 兔 

分 类 号：R743[医药卫生—神经病学与精神病学]