Geminin在血管平滑肌细胞分化表型转化中的意义探讨-第三军医大学学报.docVIP

Geminin在血管平滑肌细胞分化表型转化中的介导作用 周叶萍，舒茂琴 (400038重庆，第三军医大学西南医院心血管内科，重庆市介入心脏病学研究所) [摘要] 目的 通过比较静止状态的血管平滑肌细胞（vascular smooth muscle cells,VSMCs）与血清刺激后的VSMCs在分化表型改变中Geminin的表达变化情况，探讨Geminin对VSMCs表型转化的影响。 实验设置静止状态（血清饥饿）对照组、血清刺激组，研究不同时相2个组Geminin的分布、活性及表达水平的差异。血清饥饿法可以诱导出VSMC表型分化及活性差异；免疫荧光染色后激光共聚焦显微镜观察Geminin核内定位；应用PCR检测Geminin基因表达的变化；Western blot检测Geminin蛋白、VSMC合成型标志物骨桥蛋白（osteopontin，OPN）及收缩型标志物平滑肌肌动蛋白相关蛋白（SM-2 actin-related proteins，ARP）蛋白表达的变化。 ①静止状态下（血清饥饿）VSMC以收缩型为主，血清刺激后以合成型为主；②静止状态Geminin几乎全部存在于胞浆内，而血清刺激后Geminin几乎全部定位于胞核内；③Geminin mRNA在血清刺激组明显升高，且在刺激48h时差异表达最明显(P0.05)；④Geminin蛋白和骨桥蛋白在血清刺激组均明显升高，(P0.05)。Geminin参与介导了VSMC由收缩型向合成型的转化。 Geminin；血管平滑肌细胞；表型转化A Effect of Geminin on the phenotypic modulation of vascular smooth muscle cells of rat Zhou Yeping，Shu Maoqin (Department of Cardiology，Southwest Hospital，Third Military Medical University，Chongqing, 400038,China) [Abstract] Objective To investigate the influence of Geminin on the phenotypic modulation of vascular smooth muscle cells (VSMCs) of rat by contrast VSMCs on different states． Methods VSMCs were set up with control groups on resting state and post-stimulatory by serum, to study the disposition or activity and the difference of expression of Geminin on different phases．Serum starvation method can induce the phenotypic modulation and the difference of activity of VSMCs. The gene expression of Geminin was detected by PCR．The changes of phenotype—dependent markers and the expression of Geminin protein was analyzed by Western blot．Results The presence of control groups was mainly contractibility in endochylema, others synthesis in intranuclear. The expression of synthetic marker osteopontin and mRNA and protein of Geminin all raise(P0.05)，furthermore, the most obviously differential expression was at 48h after stimulated by serum(P0.05)．Conclusion Geminin can induce the phenotypic modulation of VSMCs from contractile to synthetic one． [Key words] Geminin；vascular smooth muscle cell；phenotypic modulation *血管平滑肌