鞘氨醇-1-磷酸对人脐带间充质干细胞增殖及表面标记物表达的影响  

Effects of Sphingosine-1-phosphate on Proliferation and Cell-surface Markers of Human Umbilical Cord Mesenchymal Stem Cells

作　　者：赵宏伟[1] 怡荣[1] 那仁格日勒[1] 高锦 陈曦[3] 阿拉坦高勒[1] 

机构地区：[1]内蒙古大学生命科学学院,呼和浩特010021 [2]北京京蒙高科干细胞技术有限公司,北京100085 [3]中国医学科学院北京协和医学院心血管病研究所,阜外心血管病医院,北京100037

出　　处：《中国生物化学与分子生物学报》2012年第5期469-476,共8页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家大学生创新性实验计划项目(No.111012612)资助~~

摘　　要：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 bioactive phospholipid molecule with multiple biological functions,such as cell proliferation,apoptosis,migration.In the present study,we found that S1P stimulated proliferation of human umbilical cord mesenchymal stem cells（hUC-MSCs）.In order to clarify the signaling pathway of hUC-MSCs proliferation in response to S1P and it＇s effect on cell-surface markers of hUC-MSCs,we detected the level of the S1P receptors（S1PRs） mRNA by real-time PCR.MTT assay was used to measure the effects of VPC23019（S1P1/3 receptor antagonist）,JTE013（S1P2 receptor antagonist）,CAY10444（S1P3 receptor antagonist）,PTX（Gi inhibitor） and PD98059（ERK inhibitor） on proliferation of hUC-MSCs induced by S1P.Then we checked the level of p-ERK by Western blotting.Flow cytometry was used to detect the cell-surface markers.The results showed that S1P stimulates hUC-MSCs proliferation in a dose dependent manner.S1PR1-3 mRNA was mainly expressed in hUC-MSCs and S1PR4,S1PR5 levels were low.VPC23019 completely and CAY10444 partly inhibited S1P-induced hUC-MSCs proliferation,but JTE013 had no significant effects.PTX and PD98059 inhibited S1P-induced hUC-MSCs proliferation and ERK1/2 phosphorylation.In conclusion,S1P stimulated the proliferation of hUC-MSCs through S1PR1/3,Gi and ERK1/2 signal pathway,but no effect was observed on the cell-surface markers of hUC-MSCs.

关 键 词：鞘氨醇-1-磷酸受体 人脐带间充质干细胞 增殖 细胞表面标记物 

分 类 号：Q25[生物学—细胞生物学]