摘要
目的:探讨同型半胱氨酸促进大鼠血管平滑肌细胞(VSMC)增殖的机制及辛伐他汀的干预作用。方法:贴壁培养的大鼠原代VSMC随机分为3组。(1)对照组:用含10%胎牛血清的DMEM培养基(完全培养基);(2)同型半胱氨酸组:在完全培养基中加入同型半胱氨酸(0.05 mmol/L);(3)辛伐他汀组:在完全培养基中加入同型半胱氨酸(0.05 mmol/L)和辛伐他汀(10μmol/L)。培养24 h后,光学显微镜下观察各组细胞的形态;采用半定量PCR法检测c-myc、P27kip1的mRNA水平;用Western blot检测细胞周期蛋白(cyclin)D1、cyclin E、cyclin A和核增殖抗原(PCNA)的蛋白表达水平。结果:与对照组相比,同型半胱氨酸组c-myc mRNA水平上调,P27kip1 mRNA水平下调,下游cyclin D1、cyclin E、cyclin A和PCNA蛋白表达增加。与同型半胱氨酸组相比,辛伐他汀组上述检测指标均得到逆转。结论:同型半胱氨酸可促进大鼠VSMC c-myc、cyclin D1、cyclin E、cyclin A等增殖相关基因的表达,抑制P27kip1表达。辛伐他汀可逆转上述效应。
Objective:To explore the mechanism of homocysteine promoting the proliferation of rat vascular smooth muscle cells (VSMC) and intervention effects of simvastatin. Methods: Primary rat VSMC were divided into 3 groups. Cells in control group were cultured in DMEM with 10% fetal bovine serum. Cells in homoeysteine group were treated with 0. 05 mmol/L homocysteine, and cells in simvastatin group were treated with 0. 05 mmol/L homocysteine and 10 μmol/L simvastatin. Morphology of cells was observed under optical microscope. The mRNA levels of c-myc 和 P27-kip1 were analyzed by semi-quantitative PCR, and the protein levels of cyclin D1, cyclin E, cyclin A and PCNA were evaluated by Western blot. Results:Compared with control group, the mRNA level of c-myc and protein levels of cyclin D1, cyclin E, cyclin A and PCNA in homocysteine group were significantly increased, while the mRNA level of P27-kip1 was decreased. After treatment with simvastatin, P27-kip1 was upregulated compared with homocysteine group accompanied by the inhibition of c-myc, cyclin D1, cyclin E, cyelin A and PCNA. Conclusion= Homocysteine promotes the expression of proliferation associated genes such as c-myc, cyelin D1, eyclin E, cyclin A and PCNA and inhibits P27-kip1 expression, which can be reversed by simvastatin.
出处
《国际心血管病杂志》
2015年第1期44-47,共4页
International Journal of Cardiovascular Disease
基金
上海市科委重大基础研究项目(054119634)