鞘氨醇1磷酸及其受体在粒细胞集落刺激因子诱导造血干/祖细胞动员过程中的表达变化

Expression changes in sphingosine-1-phosphate and its receptor during granulocyte colony stimulating factor-induced hematopoietic stem/progenitor cells mobilization

目的探讨鞘氨醇1磷酸(S1P)及其受体在粒细胞集落刺激因子(G-CSF)诱导的造血干/祖细胞(HSPC)动员中的表达变化。方法 6周清洁级雄性C57BL野生型小鼠18只,按随机数字表法分为对照组、动员3 d组、动员5 d组,每组6只。对照组给予腹部皮下注射无菌生理盐水100μl/d,连续5 d;动员3 d组连续腹部皮下注射G-CSF 3 d;动员5 d组连续腹部皮下注射G-CSF 5 d。应用流式细胞仪检测3组小鼠外周血Lin-Sca1±c Kit±(LSK)细胞水平,酶联免疫吸附实验(ELISA)检测外周血S1P水平,RT-PCR检测骨髓S1PR1 mRNA水平。结果 LSK细胞水平动员5 d组>动员3 d组>对照组(P<0.05),说明动员成功。动员3 d组外周血S1P水平明显大于动员5 d组、对照组(P<0.05),但动员5 d组与对照组比较,差异无统计学意义(P>0.05)。骨髓S1PR1 mRNA水平动员5 d组>动员3 d组>对照组(P<0.05)。结论在G-CSF诱导的HSPC动员过程中,外周血S1P水平升高以及骨髓S1PR1表达升高,可能有利于G-CSF介导HSPC动员的发生。

Objective To explore the expression changes in sphingosine-1-phosphate（S1P） and its receptor in the mobilization of granulocyte colony stimulating factor（ G-CSF）-induced hematopoietic stem/progenitor cells（HSPC）.Methods Eighteen male C57BL mice of clean grade and wild type were randomly divided into control group（n =6）,3-day mobilization group（n =6） and 5-day mobilization group（n =6）.The mice in the control group were intraperitoneally injected with 100 μl of normal saline for 5 consecutive days.The mice in the 3-day mobilization group were intraperitoneally injected with G-CSF for 3 consecutive days,and in the 5-day mobilization group for 5 consecutive days.Flow cytometry was used to detect the level of Lin-Sca1 ＋cKit ±（LSK） cells in peripheral blood,enzyme-linked immunosorbent assay（ELISA） to the level of S1P in peripheral blood,and real-time fluorescence quantitative PCR （RT-PCR） to the level of S1PR1 mRNA in bone marrow.Results The level of LSK cells was the highest in the 5-day mobilization group and the lowest in the control group（P〈0.05）.The level of S1PR1 mRNA in bone marrow was the highest in the mobilization group and the lowest in the control group（P 〈0.05）.Conclusion The increase of S1P level in peripheral blood and S 1PR1 expression in bone marrow might facilitate the G-CSF-mediated HSPC mobilization in the process of G-CSF-induced HSPC mobilization.