高糖缺氧环境下转甲状腺素蛋白对视网膜血管内皮细胞的影响

Transthyretinin repress retinal microvascular endothelial cells under high glucose and hypoxia environment

目的观察高糖缺氧环境下转甲状腺素蛋白（TTR）对人视网膜血管内皮细胞（hREC）的影响。方法分别于5．5mmol／L葡萄糖（低糖，LG）、25．0mmol／L葡萄糖（高糖，HG）以及200μmol／LCoCl2诱导的缺氧环境中培养hREC、人视网膜色素上皮细胞（hRPEC），并据此分为LG组、LG缺氧组、HG组、HG缺氧组。实时无标记细胞功能分析仪检测细胞培养初始时间（Oh）及培养后4、8、16、24、36、48、60、72h的细胞生长指数。分别在LG组、LG缺氧组、HG组、HG缺氧组细胞培养基中添加4tLmol／LTTR，即LG组＋TTR、LG缺氧组＋TTR、HG组＋TTR、HG缺氧组＋TTR，观察TTR对hREC和hRPEC的作用以及hRPEC对hREc生长的影响。Transwell共培养体系观察hPEPC对hREC生长影响。结果培养后72h，LG组hREc、hRPEC生长指数明显高于LG缺氧组、HG组，差异均有统计学意义（hREC：F=17．098、22．970，P〈0．05；hRPEC：F=45．442、9．011，P〈0．05）；HG组、HG缺氧组hREC、hRPEC生长指数显著降低，差异均有统计学意义（hREC：F=146．184，P〈0．05；hRPEC：F=27．907，P〈0．05）。HG组＋TTR、HG缺氧组＋TTRhREC生长指数显著低于HG组、HG缺氧组，两组hREC生长指数比较，差异有统计学意义（F=161．430、24．106，P〈0．05）；LG组＋TTR、LG缺氧组＋TTRhREC生长指数高于LG组，LG缺氧组，两组hREC生长指数比较，差异有统计学意义（F=200．486、48．662，P〈0．05）。LG组＋TTR、LG缺氧组＋TTR、HG组＋TTR、HG缺氧组＋TTRhRPEC生长指数均较LG组、LG缺氧组、HG组、HG缺氧组略高，但差异无统计学意义（P〉0．05）。共培养与单独生长细胞相比，共培养hRPEC对hREC生长有显著抑制作用，差异有统计学意义（LG组：F=15．711，P〈0．05；LG缺氧组：F=45．659，P〈0．05；HG组：F=7．857，P〈0．05；HG缺氧组：F=6．348，P〈0．05）。绪论高糖缺氧环境下TTR对hREC生长有抑制作用。

Objective To explore transthyretin （TTR） effect on retinal vascular endothelial cells （hREC） under high glucose and hypoxia environment. Methods hREC and human retinal pigment epithelial cell （hRPEC） were cultured at low-glucose （LG）, high glucose （HG） and hypoxia. The glucose concentration was increased from 5.5 mmol/L up to 25 mmol/L, and hypoxia was induced by 200 μmol/L CoClz. The cells were divided into LG group, LG-hypoxia group, HG group, HG-hypoxia group according to the different cell culture environment. The growth index was detected at 0, 4, 8, 16, 24, 36, 48, 60, 72 hours after cultured. Furthermore, hREC and hRPEC were also cultured with additional TTR （4μmol/L）, respectively. Then transwell co-culture system was employed to reveal the effects of hRPEC on the growth of hREC. Results At 72 hours after cultured, the growth index of hREC and hRPEC in LG group were increased as compared with LG-hypoxia group and HG group （hREC; F=17. 098, 22. 970; P〈0.05. hRPEC：F= 45. 442, 9. 011; P（0.05） ; the growth index of hREC and hRPEC were decreased in HG group and HG-hypoxia group （hREC： F= 146. 184, P〈0.05; hRPEC： F= 27. 907, P〈0.05）. Additionally, hREC could be significantly repressed by added TTR during culture with high concentration of glucose （F= 161. 430, 24. 106;P〈0.05）. hREC could be significantly increased by added TTR during culture with low concentration of glucose （F = 200. 486, 48. 662; P 〈 0.05）. In co-culture process, hRPEC revealed inhibition activity against hREC under both natural and abnormal environment （LG group.. F= 15. 711, P〈 0.05; LG-hypoxiagroup： F=45.659, P〈0.05;HG group; F=7.857, P〈0.05; HG-hypoxia group; F= 6. 348, P〈0.05）. Conclusion Under high glucose and hypoxia environment, the growth of hREC from neovascular could be inhibited by TTR.