[1]宋明宝,于学军,朱光旭,等.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[J].第三军医大学学报,2009,31(04):285-289.
 SONG Ming-bao,YU Xue-jun,ZHU Guang-xu,et al.Interaction between endothelial cells and smooth muscle cells: an in vitro study based on Transwell insert[J].J Third Mil Med Univ,2009,31(04):285-289.
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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(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
31卷
期数:
2009年第04期
页码:
285-289
栏目:
论著
出版日期:
2009-02-28

文章信息/Info

Title:
Interaction between endothelial cells and smooth muscle cells: an in vitro study based on Transwell insert
作者:
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心血管内科,全军心血管病研究所
Author(s):
SONG Ming-bao YU Xue-jun ZHU Guang-xu ZHAO Gang YU Shi-yong DONG Hong-mei KANG Hua-li HUANG Lan
Cardiovascular Diseases Research Center, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China
关键词:
内皮细胞平滑肌细胞缝隙连接共培养血管
Keywords:
endothelial cells smooth muscle cell gap junction coculture vessel
分类号:
R322.12;R329-33;R329.28
文献标志码:
A
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)调节血管平滑肌细胞(smooth muscle cell, SMC)增殖能力中的作用。    方法    采用植块法分别培养大鼠主动脉内皮及平滑肌细胞,并将2种细胞分别接种至Transwell insert膜的两侧以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2种细胞在Transwell insert膜两侧生长情况。用3H-TdR掺入法观察双层共培养模型中平滑肌细胞增殖情况,并观察缝隙连接阻断剂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1天,EC/SMC组与SMC/SMC组的平滑肌细胞3H-TdR掺入率无统计学差异[(10 900±1 320) cpm/well vs(10 430±1 200) cpm/well, P>0.05]。第2天,EC/SMC组中平滑肌细胞3H-TdR的掺入率有低于SMC/SMC组[(17 200±1 734 )cpm/well vs (20 900±1 659 )cpm/well ]的趋势,但两者差异未达到统计学标准。共培养第3天时,EC/SMC组平滑肌细胞3H-TdR掺入率显著低于SMC/SMC组,两者相比有显著性差异[(25 800±1 984 )cpm/well vs(33 070±3 569)cpm/well, P<0.05]。在用18α-GA预处理24 h后,EC/SMC共培养3 d后平滑肌细胞3H-TdR的掺入率明显高于同期未处理EC/SMC组,两者相比有显著性差异[(30 500±2 650)cpm/well vs(25 800±1 984 )cpm/well, P<0.05]。    结论    双层共培养模型中内皮细胞通过缝隙连接可抑制血管平滑肌细胞的增殖能力。
Abstract:
Objective    To explore the role of gap junctions on the interaction between endothelial cells and smooth muscle cells in a coculture system.     Methods    Rat aortic endothelial cells (RAECs) and smooth muscle cells (SMCs) were isolated and primarily cultured from rat aortic wall tissue and identified by immunofluorescence staining with vWF and smooth muscle α-actin respectively. A coculture system was established by seeding SMCs and ECs in the each side of Transwell  insert respectively and their growth was observed by transmission electron microscopy. 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 the proliferation of SMCs in the coculture system. A Transwell insert system with SMCs in both well and Transwell served as control.     Results    The ECs and SMCs grow well and attached together to form gap junctions in the coculture system. On the first day after coculture, there was no significantly difference of 3H-TdR incorporation between ECs/SMCs and SMCs/SMCs (P>0.05). On the second day, SMCs/SMCs system had a higher incorporation value of 3H-TdR than ECs/SMCs system, though there was no statistical difference.  On the third day, the incorporation value in ECs/SMCs was significantly lower than that of SMCs/SMCs system (P<0.05), suggesting that ECs suppressed SMCs proliferation. However, when SMCs in the culture well was treated with 50 μmol/L 18α-GA for 24 h, the incorporation value of 3H-TdR of ECs/SMCs system was significantly higher than that of the ECs/SMCs system without treatment (P<0.05), indicating that 18α-GA suppressed the inhibitory effect of ECs on SMCs proliferation.     Conclusion    ECs may suppress the proliferation of SMCs via gap junctions.

参考文献/References:

宋明宝, 于学军, 朱光旭, 等. 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[J]. 第三军医大学学报, 2009, 31(4):285-289.

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更新日期/Last Update: 2009-02-11