EP_2A体外可促进人CD34^+细胞归巢而不能促进其增殖  

EP_2A promotes human CD34^+ cell homing in vitro but not proliferation

作　　者：刘芳洁[1] 汪亚群[1] 唐晶[1] 陈惠珍[1] 赖淑萍[1] 苏畅[1] 李娟[1] 许多荣[1] 

机构地区：[1]中山大学附属第一医院血液科,广东广州510080

出　　处：《中国病理生理杂志》2017年第11期2026-2031,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(N0.81370663)

摘　　要：目的:探讨前列腺素E_2受体2激动剂(EP_2A)在体外对人CD34^+细胞的归巢与增殖作用。方法:收集健康供者经粒细胞集落刺激因子动员后的外周血,免疫磁珠法分选出人CD34^+细胞;同时收集健康供者动员前骨髓液,分离单个核细胞,并行骨髓间充质干细胞(BMMSC)培养。人CD34^+细胞和BMMSC经前列腺素E_2(阳性对照)、DMSO(阴性对照)、EP_2A和EP_2A+前列腺素E_2受体2拮抗剂(EP_2AA)处理后,对人CD34^+细胞用CCK-8法检测细胞活力,集落形成实验检测集落形成数目,流式细胞术检测G_2/M期细胞比例,Western blot检测细胞中survivin、β-catenin及CXC趋化因子受体4(CXCR4)的蛋白表达;ELISA法检测BMMSC中基质细胞衍生因子1α(SDF-1α)的含量。结果:EP_2A组与阴性对照组相比,人CD34^+细胞在细胞活力、集落形成数目、G_2/M期比例及survivin和β-catenin蛋白表达方面均无明显差别。但EP_2A组人CD34^+细胞CXCR4及BMMSC SDF-1α的表达均明显高于阴性对照组。结论:EP_2A体外可促进人CD34^+细胞归巢但不能促进其增殖。AIM：To investigate the role of prostaglandin E2receptor 2 agonist（EP2A）in proliferation and homing of human CD34＋cells.METHODS：Bone marrow fluid and peripheral blood containing stem cells were collected from healthy donors mobilized by granulocyte colony-stimulating factor in our department.Human CD34＋cells were isolated by the method of magnetic-activated cell sorting microbeads.Bone marrow mononuclear cells were isolated by Ficoll-Paque centrifugation,and the bone marrow mesenchymal stem cells（BMMSC）were cultured with L-DMEM.Human CD34＋cells and BMMSC were divided into 4 groups,and treated with PGE2（as positive control）,DMSO（as negative control）,EP2A and EP2A＋prostaglandin E2receptor 2 antagonist（EP2AA）,respectively.After exposed to the reagents,human CD34＋cell viability was measured by CCK-8 assay,the number of colonies was evaluated by colony-formation assay,the cell cycle distribution was analyzed by flow cytometry,and the protein expression of survivin,β-catenin and CXC chemokine receptor4（CXCR4）was detrmined by Western blot.Moreover,the concentration of stromal cell-derived factor-1α（SDF-1α）in the BMMSC was detected by ELISA.RESULTS：The cell viability and the colony number of human CD34＋cells in EP2A group were not higher than those in negative control group.Furthermore,the proportion of human CD34＋cells treated with EP2A in G2/M phase was not elevated compared with negative control group.The protein expression of survivin andβ-catenin did not up-regulated in human CD34＋cells exposed to EP2A,but the protein expression of CXCR4 in human CD34＋cells and the concentration of SDF-1αin BMMSC were elevated.CONCLUSION：EP2A promotes human CD34＋cell homing in vitro but not proliferation.

关 键 词：前列腺素E2 前列腺素E2受体2激动剂 细胞增殖 归巢 

分 类 号：R551.3[医药卫生—血液循环系统疾病] R363[医药卫生—内科学]