上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

雷帕霉素联合环孢素A抑制血管内皮细胞增殖的机制研究

尚明花1，郭海英2，王玲1，蔡敏超1，范昱3   

  1. 上海交通大学附属第一人民医院 1.肾内科， 2.呼吸科， 3.器官移植中心， 上海 200080
  • 出版日期:2015-09-28 发布日期:2015-09-30
  • 通讯作者: 范昱, 电子信箱: fanyu1965@163.com。
  • 作者简介: 尚明花（1965—），女，主任医师，硕士; 电子信箱: shangminghua@medmail.com.cn。

Mechanism of inhibiting proliferation of vascular endothelial cells by rapamycin combined with cyclosporin A

SHANG Ming-hua1, GUO Hai-ying2, WANG Ling1, CAI Min-chao1, FAN Yu3   

  1. 1.Department of Nephrology, 2.Department of Respiration, 3.Organ Transplantation Center, Shanghai First People's Hospital, Shanghai Jiao Tong University, Shanghai 200080, China
  • Online:2015-09-28 Published:2015-09-30

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摘要：

目的  探讨雷帕霉素（Rapa）对经环孢素A（CsA）诱导的血管内皮细胞增殖的抑制作用及可能的作用机制。方法  以不同浓度的Rapa干预经CsA诱导增殖的人脐静脉内皮细胞株，3H-胸腺嘧啶核苷掺入法和3H-亮氨酸掺入法测定细胞DNA和蛋白质合成，放射免疫分析法检测血管紧张素Ⅱ（AngⅡ)的分泌情况，Western blotting检测细胞信号蛋白核糖体蛋白S6激酶（p70S6K）和细胞外信号调节激酶(ERK2)的表达。结果  与CsA组比较，CsA+Rapa干预组血管内皮细胞DNA和蛋白质合成受到显著抑制，细胞培养上清液及蛋白提取液中AngⅡ的含量显著减少，且呈Rapa剂量依赖性。Western blotting检测结果显示：与CsA组比较，CsA+Rapa （10 nmol/L）干预组和CsA+Rapa （100 nmol/L）干预组p70S6K 表达明显减少；而ERK2的表达在CsA+Rapa干预组中虽呈剂量依赖性下降，但差异无统计学意义（P>0.05）。结论  Rapa可能通过抑制细胞信号蛋白p70S6K的表达，阻断PI3K-p70S6K信号通路，抑制CsA刺激的血管内皮细胞分泌AngⅡ，从而抑制血管内皮细胞的增殖。Rapa联合CsA的免疫抑制方案有利于抑制移植物血管内皮细胞的增殖。

关键词: 雷帕霉素, 核糖体蛋白S6激酶, 环孢素A, 血管紧张素Ⅱ

Abstract:

Objective  To investigate the inhibitory effect of rapamycin (Rapa) on the proliferation of vascular endothelial cells induced by cyclosporin A (CsA) and possible mechanisms. Methods  Human umbilical vein endothelial cells (HUVECs) that proliferated under the induction of CsA were treated by different concentrations of Rapa. The synthesis of DNA and protein was detected by 3H-TdR incorporation and 3H-leucine incorporation. The secretion of Ang Ⅱ in cytoplasm and culture medium was measured by radioimmunoassay. Western blotting was adopted to assess expressions of ribosomal protein S6 kinase (p70S6K) and ERK2. Results  Compared with the CsA group, the synthesis of DNA and protein of vascular endothelial cells of the CsA+Rapa intervention group was significantly inhibited and AngⅡ levels in culture supernatant and protein extract significantly decreased and were dose-dependent with Rapa. Results of Western blotting showed that compared with the CsA group, the expressions of p70S6K of the CsA+Rapa (10 nmol/L) intervention group and CsA+Rapa (100 nmol/L) intervention group significantly decreased. The expression of ERK2 of the CsA+Rapa intervention group decreased in a dose-dependent manner, but the difference was not statistically significant (P>0.05).  Conclusion  Rapa may inhibit expression of p70S6K, block PI3K-p70S6K signaling pathway, and inhibit the vascular endothelial cells stimulated by CsA from secreting AngⅡ, so as to inhibit the proliferation of vascular endothelial cells by inhibiting the expression of signal protein p70S6K. The immunosuppression therapy of Rapa combined with CsA facilitates inhibiting the proliferation of vascular endothelial cells of grafts.

Key words: rapamycin, ribosomal protein S6 kinase, cyclosporine A, angiotensin Ⅱ