1-磷酸鞘氨醇受体2在LPS介导的内皮细胞高通透性中的作用

Effect of Sphingosine 1-Phosphate Receptor 2 on Endothelial Hyper-Monolayer Permeability Induced by LPS

目的观察1-磷酸鞘氨醇受体2(S1PR2)在脂多糖(LPS)刺激后表达的变化,探讨其在LPS介导的内皮细胞高通透性中的作用。方法用不同浓度和时间的LPS刺激培养的ECV304细胞,采用RT-PCR方法检测S1PR2mRNA的变化,Western blot方法检测S1PR2蛋白的变化,TRITC荧光标记白蛋白漏出法测定单层内皮细胞的通透系数Pa值。结果与对照组相比,S1PR2mRNA和S1PR2蛋白在LPS(500ng/ml)刺激24h后以及在LPS(1000ng/ml)刺激12h显著增加(P<0.01);S1PR2拮抗剂JTE-013能显著降低LPS引起的内皮细胞通透性增高(P<0.05)。结论 S1PR2在LPS引起的内皮细胞通透性增高中起着重要的作用。

Objective To study the changes of expression level of sphingosine 1-phosphate receptor 2 （S1PR2） and the effects of S1PR2 activation in hyper-monolayer permeability in ECV 304 under the stimulation of LPS. Methods Different doses of LPS were co-cultured with ECV 304 for different periods. RT-PCR was used for semiquantitative measurement of the change of the S1PR2 mRNA expression. Western blot was used to detect the change of S1PR2 protein expression. ECV 304 monolayer permeability was detected by measuring the leakage of albumin labeled with fluorescein isothiocyanate （FITC） in the lower compartment of transwell. Pa value reflects the permeability of endothelial monolayer. Results There were significant increases in S1PR2 mRNA expression 24 h after stimulated by LPS （500 ng / ml） and 12 h after stimulated by LPS （1 000 ng / ml）compared with control （P0.01）. Consistent with the RT-PCR result,S1PR2 protein expression increased remarkably after LPS application. S1PR2 inhibitor （JTE-013） attenuated the hyper-monolayer permeability under stimulation of LPS （P0.05）. Conclusion The upregulation of S1PR2 after LPS administration suggested that S1PR2 might play an important role in LPS-induced hypermonolayer permeability in ECV 304.