巨细胞病毒感染对定向干细胞增殖的影响及黄芪的干预作用

Effect of cytomegalovirus infection on proliferation of committed stem cells with the presence of Astragalus Membranaceus injection in vitro

目的:探讨巨细胞病毒(CMV)感染对定向干细胞(CFU-Gm、CFU-E、BFU-E、CFU-Mk及CFU-TL)体外增殖抑制的影响及黄芪的干预作用。方法:采用造血细胞培养技术,培养、观察、计数CMV-AD169感染CFU-Gm、CFU-E、BFU-E、CFU-Mk及CFU-TL后各祖细胞集落及维持时间;用聚合酶链反应(PCR)和荧光定量PCR技术检测集落细胞内CMV-AD169DNA;根据细胞毒性实验结果,将黄芪作用于CMV感染的CFU-Gm、CFU-E、BFU-E、CFU-Mk及CFU-TL祖细胞,再分别计数集落、峰期及维持时间。结果:(1)CMV感染组CFU-Gm、CFU-E、BFU-E、CFU-Mk及CFU-TL集落数较对照组明显减少(P<0.01);集落维持时间CMV感染组较对照组明显缩短;(2)用PCR在感染组集落细胞内检测到HCMV-DNA的存在;(3)黄芪作用于CMV感染的定向干细胞后,与病毒组比较集落数明显提高(P<0.01),集落增殖提高率分别为CFU-GM27.2%;CFU-E45.2%;BFU-E49.1%;CFU-Mk11.9%,CFE-TL55.2%,且集落维持时间明显延长;(4)荧光定量PCR技术检测集落细胞内CMV-DNA复制较CMV感染组明显减少(P<0.01)。结论:CMV-AD169株在体外能明显抑制CFU-Gm、CFU-E、BFU-E、CFU-Mk及CFU-TL集落细胞的增殖;在体外黄芪有明显抗CMV作用,能促进CMV感染的定向干细胞增殖。

OBJECTIVE To investigate the effect of Astragalus Mernbranaceus injection（AMI） on suppression proliferation of colony forming gunitgranulocyte-macrophage （CFU-Gm）, CFU-Erythroid （CFU-E）, burst forming unit-erythroid （BFU-E）. CFU-megakaryocyte （CFU-Mk） and CFU-T lymphocyte（CFU-TL） progenitor cells of cord blood（CB） with cytomegalovirus （CMV） infection. METHODS 20 CB samples were collected from fetal umbilical vein of normal term spontaneous delivery neonates. Colony forming unit-assay was applied to observe the suppression effect of CMV-AD169 strain on CFU-Gm, CFU-E, BFU-E, CFU-Mk and CFU-TL of CB with the presence of AMI in vitro. The polymerase chain reaction （PCR） and fluorescence quantification PCR were used to demonstrate the existence of CMV-AD169 DNA in the colony cells of cultured hematopoietic progenitors. RESULTS The numbers of CFU-GM,CFU-E, BFU-E,CFU-Mk and CFU-TL colonies in CMV infection groups were significantly less than that in blank and mock group respectively. The maintaining duration, of colonies in groups with CMV infection was significantly shorter compared to blank and mock. CMV-DNA was positively detected in the colony cells of CMV infection groups by PCR,while negative in the control groups. The maintaining duration, of CFU-GM,CFU-E, BFU-E, CFU-Mk and CFU-TL colonies infected with CMV extended significantly with the presence of AMI,and the numbers of those increased significantly compared with CMV infection group respectively. The proliferation rate of colonies was 27.2%,45.20%, 49. 1%,55.1%,11.9% with AMI CMV groups respectively. PCR amplification demonstrated the existence of HCMV-AD169 DNA in the colony cells of the Infection groups. CMV-AD169 DNA copies decreased obviously by fluorescence quantification PCR in HCMV group with the presence of AMI. CONCLUSION CMV-AD169 strain can infect hematopoietic progenitors of cord blood and inhibit the proliferation of hematopoietic progenitors. The growth of hematopoietic progenitor cell after CMV AD169 infection is promoted by AMI,which suggests that AMI exert an effect of anti-CMV in vitro.