血小板生成素通过修复化疗后骨髓内皮祖细胞促进巨核细胞造血  

Thrombopoietin promotes megakaryopoiesis via protecting bone marrow endothelial function in patients undergoing chemotherapy for hematological malignancies

作　　者：曾晓媛 焦营营 李宗鹏 张玉娇 叶洁瑜[1] ZENG Xiaoyuan;JIAO Yingying;LI Zongpeng;ZHANG Yujiao;YE Jieyu(Department of Hematology,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China)

机构地区：[1]南方医科大学南方医院血液科,广东广州510515

出　　处：《南方医科大学学报》2020年第8期1134-1140,共7页Journal of Southern Medical University

基　　金：广东省自然科学基金(217A030313767)。

摘　　要：目的探索血小板生成素(TPO)能否通过修复大剂量化疗后患者的骨髓内皮祖细胞(BM-EPC)恢复其对巨核细胞的造血支持作用。方法选取23例血液系统恶性肿瘤患者化疗后30 d的骨髓以及10例健康人骨髓作为实验标本。采用表面特异性抗原CD34,CD309,CD133对BM-EPC进行鉴定;使用CCK8分析TPO是否可促进血液肿瘤患者化疗后BM-EPC增殖及其最佳作用浓度。设置TPO处理组为实验组,无TPO为对照组,健康人BM-EPC为健康对照组;通过DiL-Ac-LDL摄取及FITC-UEA-I结合实验检测实验组、对照组和健康对照组的BM-EPC数量。利用成管及迁移实验评估3组的BM-EPC功能;分别将实验组和对照组BM-EPC与巨核细胞共培养,利用流式细胞术检测巨核细胞增殖情况。结果实验组BM-EPC高表达CD34/CD133/CD309;TPO可促进BM-EPC增殖,最佳作用浓度为100μg/L。免疫荧光双标实验显示,实验组BM-EPC数量较对照组明显增加(P<0.05)。实验组成管能力及迁移能力较对照组增强(P<0.05)。共培养后,实验组巨核细胞数多于对照组(P=0.013)。结论大剂量化疗后患者BM-EPC受损,TPO具有直接刺激巨核系统造血作用,还可能通过促进BM-EPC增殖,修复其功能,从而恢复BM-EPC对巨核细胞的造血支持作用。Objective To explore whether thrombopoietin(TPO)can rescue megakaryopoiesis by protecting bone marrowderived endothelial progenitor cells(BM-EPCs)in patients receiving chemotherapy for hematological malignancies.Methods Bone marrow samples were collected from 23 patients with hematological malignancies 30 days after chemotherapy and from 10 healthy volunteers.BM-EPCs isolated from the samples were identified by staining for CD34,CD309 and CD133,and their proliferation in response to treatment with TPO was assessed using CCK8 assay.DiL-Ac-LDL uptake and FITC-UEA-I binding assay were performed to evaluate the amount of BM-EPCs from the subjects.Tube-formation and migration experiments were used for functional assessment of the BM-EPCs.The BM-EPCs with or without TPO treatment were co-cultured with human megakaryocytes,and the proliferation of the megakaryocytes was detected with flow cytometry.Results Flow cytometry indicated that the TPO-treated cells had high expressions of CD34,CD133,and CD309.CCK8 assay demonstrated that TPO treatment enhanced the proliferation of the BM-EPCs,and the optimal concentration of TPO was 100μg/L.Double immunofluorescence assay indicated that the number of BM-EPC was significantly higher in TPO-treated group than in the control group.The TPO-treated BM-EPCs exhibited stronger tube-formation and migration abilities(P<0.05)and more significantly enhanced the proliferation of co-cultured human megakaryocytes than the control cells(P<0.05).Conclusion TPO can directly stimulate megakaryopoiesis and reduce hemorrhage via protecting the function of BM-EPCs in patients following chemotherapy for hematological malignancies.

关 键 词：血小板生成素 骨髓内皮祖细胞 巨核细胞造血 

分 类 号：R730.53[医药卫生—肿瘤]