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鞘氨醇-1-磷酸(sphingosine-1-phosphate,S1P)是一种脂质信号分子,与细胞增殖、凋亡和迁移等有密切关系.本研究发现,S1P促进人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hUC-MSCs)增殖,但目前关于其作用信号通路及S1P对hUC-MSCs表面标记表达的影响尚不十分清楚.Real-time PCR检测hUC-MSCs中S1P受体mRNA表达情况,发现在hUC-MSCs中优势表达S1PR1-3,而S1PR4、S1PR5的表达很少.MTT法检测S1PR1/3拮抗剂VPC23019、S1PR2拮抗剂JTE013、S1PR3拮抗剂CAY10444、Gi蛋白抑制剂PTX和ERK抑制剂PD98059对S1P诱导hUC-MSCs增殖的影响.结果显示,VPC23019完全抑制S1P诱导的hUC-MSCs增殖,JTE013对此没有明显影响,CAY10444部分抑制S1P诱导的hUC-MSCs增殖,PTX、PD98059完全抑制S1P诱导hUC-MSCs增殖.进一步用Western印迹检测ERK1/2磷酸化水平揭示,S1P通过促进ERK1/2磷酸化进而促进hUC-MSCs增殖.流式细胞术检测发现,S1P对hUC-MSCs表面标记物(CD45、CD34、CD90、CD29、CD105、CD44、CD73、CD71)表达没有明显影响.本研究证明,S1P通过S1PR1/3、Gi偶联蛋白及ERK1/2信号通路促进hUC-MSCs增殖,而对hUC-MSCs表面标记物表达无明显影响.
Sphingosine-1-phosphate (S1P) is a lipid signaling molecule that is closely related to cell proliferation, apoptosis, migration, etc. The present study found that S1P promotes human umbilical cord mesenchymal stem cells However, the role of signaling pathway and S1P on the surface marker expression of hUC-MSCs is not yet clear.Real-time PCR detection of S1P receptor mRNA expression in hUC-MSCs S1PR1 / 3 antagonist VPC23019, S1PR2 antagonist JTE013, S1PR3 antagonist CAY10444, Gi protein inhibitors PTX and ERK were found to be predominantly expressed in hUC-MSCs, but not S1PR4 and S1PR5 Inhibitory effect of inhibitor PD98059 on proliferation of hUC-MSCs induced by S1P The results showed that VPC23019 completely inhibited the proliferation of hUC-MSCs induced by S1P, but JTE013 had no obvious effect on it. CAY10444 partially inhibited the proliferation of hUC-MSCs induced by S1P, and inhibited PTX and PD98059 completely S1P induced proliferation of hUC-MSCs. Further examination of the phosphorylation level of ERK1 / 2 by Western blot revealed that S1P promoted the proliferation of hUC-MSCs by promoting the phosphorylation of ERK1 / 2. Flow cytometry revealed that S1P inhibited the proliferation of hUC-MSCs surface marker (CD45, The results showed that S1P promoted the proliferation of hUC-MSCs via S1PR1 / 3, Gi coupled protein and ERK1 / 2 signaling pathway, but not hUC-MSCs Surface marker expression had no significant effect.