Effects of rapamycin on the epithelial-to-mesenchymal transition of human peritoneal mesothelial cells
Effects of rapamycin on the epithelial-to-mesenchymal transition of human peritoneal mesothelial cells
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DOI：
10.1177/039139880502800213
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发表时间：
2005-02-01
影响因子：
1.7
通讯作者：
Selgas, R
中科院分区：
文献类型：
--
作者：
Aguilera, A;Aroeira, LS;Selgas, R
腹膜的保存对腹膜透析患者的长期生存至关重要。上皮-间充质转化（Epithelial-to-mesenchymal transition， EMT）是间充质细胞（mesoepithelial cells， MC）的一个过程，负责腹膜的阴性变化，与PD直接相关。EMT促进了MC的新生血管和纤维化能力。血管内皮生长因子（VEGF）是新生血管形成的介质。雷帕霉素是一种有效的免疫抑制剂，在同种异体肾移植中具有抗纤维化作用，并具有抗vegf的作用。我们假设雷帕霉素可以调节mct的EMT。当间皮细胞到达汇合处时，用r- tgf - β (1 ng/mL)刺激部分间皮细胞，诱导与雷帕霉素（0.2、2、4、20和40 nM）共给药的EMT。其他细胞组分别接受相似剂量的雷帕霉素或r- tgf - β。分别于6、24、48小时和7天对细胞进行分析。作为EMT的标志物，我们采用定量RT-PCR方法包括α - sma、e -钙粘蛋白和蜗牛核因子。EMT标记物和调节因子在雷帕霉素作用下表现出以下变化：e -钙粘蛋白（EMT的一种保护基因）在40 nM下的24h较对照组增加2.5倍。重要的是，雷帕霉素在6小时抑制了tgf - β诱导的蜗牛表达，而tgf - β使蜗牛的表达增加了10倍。第7天，雷帕霉素无抗emt作用。在加入雷帕霉素后，MC中α - sma的表达显著降低。综上所述，雷帕霉素对EMT表现出轻微的保护作用，因为它增加了E-cadherin的表达，降低了a- sma的表达。因此，雷帕霉素可能部分调节间皮细胞从上皮细胞向间质细胞的转变。
The preservation of the peritoneal membrane is crucial for long-term survival in peritoneal dialysis. Epithelial-to-mesenchymal transition (EMT) is a process demonstrated in mesothelial cells (MC), responsible for negative peritoneal changes and directly related to PD. EMT enables neovascularization and fibrogenic capabilities in MC. Vascular endothelial growth factor (VEGF) is the mediator for neo-vascularization. Rapamycin is a potent immunosuppressor with antifibrotic action in renal allografts and has a demonstrated anti-VEGF effect. We performed this study with the hypothesis that rapamycin may regulate the EMT of MC.MC from human omentum were cultured. When mesothelial cells reached confluence, some of them were stimulated with r-TGF-beta (1 ng/mL) to induce EMT co-administered with rapamycin (0.2, 2, 4, 20 and 40 nM). Other groups of cells received similar doses of rapamycin or r-TGF-beta, separately. Cells were analyzed at 6, 24, 48 hours and 7 days. As markers of EMT we included alpha-SMA, E-cadherin and snail nuclear factor by quantitative RT-PCR.EMT markers and regulators demonstrated the following changes with rapamycin: E-cadherin (a protective gene for EMT) increased 2.5-fold relative to controls under 40 nM, at 24h. Importantly, rapamycin inhibited snail expression induced by TGF-beta at 6h, whereas TGF-beta increased snail 10-fold. At day 7, rapamycin showed no anti-EMT properties. An important decrease in alpha-SMA expression by MC after rapamycin addition was observed.In conclusion, rapamycin shows a mild protective effect on EMT, as it increases E-cadherin and decreases a-SMA expression. Consequently, rapamycin might partially regulate the epithelial-to-mesenchymal transition of mesothelial cells.