S1PR1激活PI3K/Akt保护高糖环境下视网膜血管内皮细胞的生物学功能

S1PR1 protects the biological function of retinal vascular endothelial cells under high glucose condition via ac⁃tivating PI3K/Akt

目的研究1⁃磷酸鞘氨醇受体1(S1PR1)对高糖环境下人视网膜血管内皮细胞(HREC)的迁移、成管能力的影响,并进一步探讨其作用机制。方法将HREC随机分为空白病毒组、S1PR1过表达组、高糖+空白病毒组和高糖+S1PR1过表达组。采用细胞划痕及Transwell实验检测细胞迁移能力,成管实验检测细胞成管能力,并采用Western blot检测PI3K/Akt通路相关蛋白分子表达情况。结果与空白病毒组相比,高糖+空白病毒组的伤口愈合百分比、细胞迁移数量、成管分支点数及小管总长度均显著减少(均P<0.01);而与高糖+空白病毒组相比,高糖+S1PR1过表达组均显著增多(均P<0.01)。各组间PI3K、Akt总蛋白表达差异均无统计学意义均无显著差异(均P>0.05),而p⁃PI3K、p⁃Akt蛋白表达差异均有统计学意义均具有显著性差异(均P<0.01);与空白病毒组相比,高糖+空白病毒组的p⁃PI3K、p⁃Akt均显著减少(均P<0.01);而与高糖+空白病毒组相比,高糖+S1PR1过表达组均显著增多(均P<0.01)。结论S1PR1通过激活PI3K/Akt通路,保护高糖环境下HREC的迁移及成管能力。

Objective To study the effect of sphingosine 1⁃phosphate receptor 1(S1PR1)on the migration and tube formation of human retinal vascular endothelial cells(HREC)under high glucose condition,and to further explore its mechanism.Methods A prospective case⁃control study.HREC were randomly divided into control group,S1PR1 overex⁃pression group,high glucose+control group and high glucose+S1PR1 overexpression group.Cell scratch assays and tran⁃swell were used to detect the migration of HREC,tube formation assay was used to detect the tube formation of HREC,and Western blot was used to detect the expression of PI3K/Akt pathway⁃related protein molecules.Results Compared with the control group,the wound healing percentage,the cell number of migration,the number of branch points and the total length of the tubules in the high glucose+control group were significantly reduced(all P<0.01).While compared with the high glucose+control group,there were significantly increased in the high glucose+S1PR1 overexpression group(all P<0.01).There were no significant differences in the total protein expressions of PI3K and Aktamong the groups(all P>0.05),but there were significantly different in the expressions of p⁃PI3K and p⁃Akt(all P<0.01).Compared with the control group,the expressions of p⁃PI3K and p⁃Akt in the high glucose+control group were significantly decreased(all P<0.01).While compared with the high glucose+control group,their expressions in the high glucose+S1PR1 overex⁃pression group were significantly increased(all P<0.01).Conclusions S1PR1 protects the migration and tube⁃formation ability of HREC under high glucose condition by activating the PI3K/Akt pathway.