骆驼蓬水提物对高糖作用下视网膜血管内皮细胞的调控作用及其机制

Effects and mechanism of Luotuopeng's aqueous extract on the high glucose induced-retinal vascular endothelial cells

目的探讨骆驼蓬水提物对高糖作用的视网膜血管内皮细胞的调控作用及其对Hippo信号通路的影响。方法将视网膜血管内皮细胞分为5组:正常组(葡萄糖浓度为5. 5 mmol/L)、模型组和骆驼蓬高、中、低剂量组(葡萄糖浓度为25 mmol/L),骆驼蓬高、中、低剂量组再分别加入250、200、150 mg/L骆驼蓬水溶液。通过CCK-8法测定各组细胞的细胞活力,通过Transwell法检测细胞迁移数目,Western Blot法检测各组细胞TEA结构域转录因于1(TEAD1)与大肿瘤抑制基因1(Lats1)蛋白表达水平。结果与正常组相比,模型组细胞活力增加,迁移数目增多;与模型组相比,骆驼蓬不同剂量均可降低细胞活力、抑制细胞迁移(P <0. 05),差异有统计学意义。与正常组相比,模型组中TEAD1的蛋白表达水平升高,Lats1的蛋白表达水平降低;与模型组比较,骆驼蓬不同剂量组TEAD1蛋白表达水平降低,Lats1的蛋白表达水平升高(P <0. 05),差异有统计学意义。结论骆驼蓬可能通过调节Hippo信号通路阻碍高糖作用下的视网膜血管内皮细胞的细胞活力与迁移能力,从而调节视网膜血管内皮细胞的功能。

Objective To investigate the effects of Luotuopeng（ Herb of Common Peganum,Peganum hannal L,LTP） aqueous extract on the high glucose induced-retinal vascular endothelial cells and the molecular mechanism. Methods The retinal vascular endothelial cells,were divided into 5 groups：normal group（ 5. 5 mmol/L glucose）,model group（ 25 mmol/L glucose）,high-dose LTP group（ 25 mmol/L glucose plus 250 mg/L LTP solution）,mid-dose LTP group（ 25 mmol/L glucose plus 200 mg/L LTP solution）,and low-dose LTP group（ 25 mmol/L glucose plus 150 mg/L LTP solution）. After 3 days of incubation,cell viability was determined with CCK-8 assay. The transwell assay was performed to observe the amount of cell migration. Western Blot assay was used to measure the protein expression level of TEAD 1 and Lats 1. Results CCK-8 assay showed that compared with the normal group,the model group had promoted the cell viability of RF/6A（ retinal vascular endothelial cells）. In comparison with the model group,LTP＇s aqueous extract at different doses had all caused a lower cell viability. Transwell results proved that compared with the normal group,the model group had facilitated the ability of migration of RF/6A. In comparison to the model group,LTP＇s aqueous extract at different doses had significantly inhibited the migration ability of RF/6A（ P 〈0. 05）. Western Blot assay indicated that compared with the normal group,the model group had showed a lower expression level of TEAD1 and a higher expression level of Lats1. In comparison to the normal group,LTP＇s aqueous extract at different doses had decreased the expression of TEAD1 and increased the expression level of Lats1（ P 〈0. 05）.Conclusion LTP ＇s aqueous extract can inhibit the cell viability and migration of retinal vascular endothelial cells by regulating the Hippo signal pathway,which accounts for its regulatory effects on retinal vascular endothelial cells.