β-淀粉样蛋白对体外培养的血管平滑肌细胞的作用及同型半胱氨酸对其影响

Effects of homocysteine on cultured vascular smooth muscle cells from β-amyloid protein

目的观察同型半胱氨酸(Hcy)对受β-淀粉样蛋白1-40(Aβ1-40)作用的体外培养的血管平滑肌细胞(VSMC)的影响。方法取大鼠主动脉段,体外进行培养,分别加入不同浓度Aβ1-40和同型半胱氨酸,光镜下观察细胞数量及形态,利用MTT法观察细胞增殖及细胞活力,流式细胞技术检测观察细胞凋亡情况,SOD以及H2O2试剂盒分别检测细胞超氧化物歧化酶的活力以及过氧化氢产生情况。结果(1)光镜下观察可见,Aβ1-40、Hcy与血管平滑肌细胞共同培养时细胞较Aβ1-40单独作用时明显减少;(2)MTT结果显示Aβ1-40抑制体外培养的血管平滑肌细胞的增生并可致其凋亡,在有同型半胱氨酸存在的时候Aβ1-40对血管平滑肌细胞增殖的抑制作用以及促进凋亡作用都有增加,此作用具有时间及浓度依赖性;(3)SOD活力以及H2O2检测示Aβ1-40、Hcy共同作用时SOD活力以及H2O2含量较Aβ1-40单独作用时明显增加。结论同型半胱氨酸对于β-淀粉样蛋白1-40所致的体外培养血管平滑肌细胞损伤有促进作用。

Objective To study the effects of homocysteine （Hcy） on cultured vascular smooth muscle cells （VSMC） from β-amyloid protein 1-40（Aβ1-40）. Methods Vascular smooth muscle cells （VSMC） were isolated from the aortic vessels of rats and cultured in vitro. The Aβ1-40 and Hcy was added to the VSMC medium to observe the viability and proliferation of VSMC by MTT and flow cytometric examination, the SOD and H2O2 kit were also used to detect cell superoxide dismutase, as well as the vitality of the situation of hydrogen peroxide generated. Results （1）Observed under light microscope, the cells more significantly reduced when Aβ1-40, Hcy and VSMC co-cultured than Aβ1-40 cultured with VSMC. （2）Aβ1-40 could significantly inhibit the proliferation of VSMC and induce its apoptosis. Hcy strengthened cell damage and increased apoptosis induced by Aβ of cultures VSMC in a dose and time dependent manner. （3）The detect of SOD dynamic and H2O2 concentration showed that the combined effect of Aβ1-40 and Hcy was stronger than the effect of Aβ1-40 alone. Conclusions Homocysteine （Hcy） could increase cultured vascular smooth muscle cells from Aβ1-40 induced cytotoxicity.