Tumstatin肽对视网膜微血管内皮细胞迁移及P38MAPK蛋白表达的影响

目的:观察tumstatin肽对体外培养的视网膜微血管内皮细胞迁移及P38MAPK蛋白表达的影响,初步探讨tumstatin肽抗视网膜内皮细胞迁移的机制。方法:采用细胞划痕实验测定tumstatin肽(T8肽)对血管内皮生长因子(VEGF)诱导下RF/6A细胞(恒河猴视网膜微血管内皮细胞)迁移的影响;Western blotting检测T8肽对VEGF刺激后15,30,45,60min的RF/6A细胞P38MAPK蛋白水平的变化。结果:Tumstatin肽对RF/6A细胞迁移具有抑制作用,且可抑制VEGF对RF/6A细胞的促迁移作用,呈剂量依赖性。正常情况下,RF/6A细胞无P38MAPK蛋白的表达,但VEGF可诱导其表达P38MAPK蛋白,而tumstatin可抑制VEGF诱导的RF/6A细胞P38MAPK蛋白的表达(加入20mg/LT8肽30,45,60min时蛋白表达受到显著抑制,差异有显著性意义,P<0.01)。结论:Tumstatin抑制视网膜微血管内皮细胞的迁移,其作用可能与P38MAPK通路有关。

The effect of tumstatin peptide on the migration and expression of P38MAPK protein inretinal microvascular endothelial cells

·AIM:To observe the effect of tumstatin peptide on the migration and expression of P38MAPK protein in retinal microvascular endothelial cells in vitro..METHODS:Cell scratch healing assay was used to examine tumstatin’s interfering effect on the migration induced by vascular endothelial growth factor(VEGF)in RF/6A cells(cell line of retinal microvascular endothelial cell of Macaca rhesus).The expression of P38MAPK protein was examined by Western blotting test at 15,30,45 and 60 minutes after VEGF was stimulated by tumstatin peptide..RESULTS:Tumstatin peptide can inhibit the migration of RF/6A cells directly and can also inhibit the up-regulated migration of RF/6A cells induced by VEGF.It was in dose-dependent manner.There was no expression of P38MAPK protein in RF/6A cells in normal conditions,and VEGF could stimulate RF/6A cells to express P38MAPK protein,but tumstatin can inhibit the expression of P38MAPK protein induced by VEGF in RF/6A cells..CONCLUSION:Tumstatin can inhibit the migration of retinal microvascular endothelial cells.The inhibitory effect may be related to the signal path through P38MAPK.P38MAPK is not the only pathway;the knob of inhibition may be located in the upstream of P38MAPK..