双层共培养模型中内皮细胞对平滑肌细胞增殖能力-第三军医大学学报.DOCVIP

文章编号： 双层共培养模型中缝隙连接在内皮细胞调节平滑肌细胞增殖能力中的作用 宋明宝，于学军，朱光旭，赵 刚，于世勇，董红梅，康华利，黄 岚 （第三军医大学新桥医院全军心血管病研究所，重庆 400037） 摘 要：目的 探讨在双层共培养体系中缝隙连接在血管内皮细胞（endothelial cell, EC）本实验采用植块法分别培养大鼠主动脉内皮及平滑肌细胞，建立内皮细胞与平滑肌细胞双层共培养模型，并用透射电镜观察种细胞在transwell insert膜两侧生长情况3H-TdR掺入法观察增殖情况，18α-甘草次酸（18α-GA）对平滑肌细胞增殖能力的影响。结果 在共培养第天，EC/SMC组SMC/SMC组平滑肌细胞3H-TdR掺入无统计学差异(10900±1320) cpm/well vs.（10430±1200） cpm/well, P＞0.05。第天，EC/SMC组平滑肌细胞3H-TdR的掺入SMC/SMC组(17200±1734 )cpm/well vs. （20900±1659 ）cpm/well ]的趋势,但两者差异未达到统计学标准。第天，EC/SMC组平滑肌细胞3H-TdR掺入SMC/SMC组两者相比有显著性差异(25800±1984 )cpm/well vs.（ 33070±3569） cpm/well, P＜0.05。在用18α-GA预处理24h后，EC/SMC共培养后平滑肌细胞3H-TdR的掺入明显高于未处理EC/SMC组，两者相比有显著性差异(30500±2650)cpm/well vs. （25800±1984 ）cpm/well, P＜0.05。 关键词：内皮细胞；平滑肌细胞；缝隙连接；共培养；血管 中图法分类号： 文献标识码：A Role of gap junctions on interactin between endothelial cells and smooth muscle cell in a coculture system SONG Ming-bao, YU Xue-jun, ZHU Guang-xu,, ZHAO Gang，YU Shi-yong, DONG Hong-mei, KANG Hua-li，HUANG Lan (Institute of Cardiovascular Diseases of PLA, Xinqiao Hospital, Third Military Medical University, Chongqing 400037) Abstract: Objective To explore the Role of gap junctions on interactin between endothelial cells and smooth muscle cell in a coculture system. Methods and Materials Rat aortic endothelial cells (RAECs) and smooth muscle cells (SMCs) were cultured by explanted rat aortic wall tissue and were identified with cell immunofluorescence staining for vWF or smooth muscle α-actin. A coculture system was made by seeding ECs and SMCs on the both side of transwell insert with millipores respectively and observed with transmission electron microscope. Then, the proliferation of SMCs was evaluated by 3H-TdR incorporation. Meanwhile, 18α-glycyrrhetinic acid (18α-GA), a specific blocker of gap junction, was administered to observe its effect on vascular smooth muscle cell proliferation in the coculture system. Results At first day after coculture, there was no significantly differenc