妊娠期齐多夫定干预对新生儿、新生鼠造血干/祖细胞活性的影响

Influence of zidovudine administration during pregnancy on the activity of haematopoietic stem/progenitor cells in neonatus and neonatal mice

目的:探讨齐多夫定(AZT)对新生儿、新生鼠造血干/祖细胞活性的影响。方法:取孕期接受高效抗逆转录病毒治疗(HAART,AZT+3TC+LPV/r)的HIV感染孕产妇新生儿脐带血15份,以及正常新生儿脐带血12份,分离脐血单个核细胞,流式细胞术检测CD34^+细胞比例,培养粒单系祖细胞(CFU-GM)、红系祖细胞(BFU-E)以及巨核系祖细胞(CFU-Meg),计算克隆形成集落数。取脐血造血干/祖细胞、新生小鼠骨髓单个核细胞,在CFU-GM、BFU-E和CFU-Meg体系和造血干/祖细胞悬浮培养体系中,进行体外AZT干预,计算成集落数及凋亡率。取Balb/c孕小鼠,进行宫内AZT(5 mg、10 mg)干预,检测新生小鼠骨髓CFU-GM、BFU-E和CFU-Meg成集落数。结果:HAART暴露的脐血单个核细胞中,CD34^+造血干/祖细胞比例、CFU-GM、BFU-E和CFU-Meg集落数均显著低于正常脐血(P<0.05)。脐血造血干/祖细胞和新生小鼠骨髓细胞在1.0μmol/L AZT干预下,CFU-GM、BFU-E和CFU-Meg成集落数显著降低(P<0.05);AZT浓度>0.5μmol/L时,脐血造血干/祖细胞和新生小鼠骨髓细胞凋亡率显著升高(P<0.05)。宫内不同剂量AZT暴露的新生小鼠骨髓CFU-GM、BFU-E和CFU-Meg集落数明显低于正常对照组(P<0.01)。结论:妊娠期AZT干预可明显抑制造血干/祖细胞的活性。

Objective:To investigate the influence of zidovudine(AZT)administration during pregnancy on the activity of haematopoietic stem/progenitor cells(HSPCs)in neonatus and neonatal mice.Methods:Cord blood from 15 HIV mothers(received highly active antiretroviral therapy,HAART,AZT+3TC+LPV/r)and 12 HIV-uninfected mothers were collected.The proportion of CD34 cells was detected by flow cy-tometry.The formation of HSPCs colonies including granulocyte-monocyte colony forming units(CFU-GM),burst forming unit-erythroid(BFU-E)and colony forming unit-megakaryocyte(CFU-Meg)was cal-culated.Umbilical cord blood HSPCs and bone marrow cells of neonatal mice exposed to AZT were isolated and cultured in vitro.The CFU-GM,BFU-E and CFU-Meg colonies and the apoptosis rate of HSPCs were observed.Results:HAART exposure resulted in a decreased rate of CD34 cells,and decreased number of colonies including CFU-GM,BFU-E and CFU-Meg(P<0.05).After treated with 1.0umol/L AZT,the number of colonies including CFU-GM,BFU-E and CFU-Meg were significantly reduced(P<0.05).The apoptosis rate in cord blood HSPCs and bone marrow cells of neonatal mice was increased by treatment of AZT(>0.5umol/L).Administration of AZT at different doses during pregnancy could inhibit the colony formation of CFU-GM,BFU-E and CFU-Meg(P<0.01).Conclusion:Administration of AZT during pregnancy could suppress the activity of HSCPs.