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目的探讨人巨细胞病毒(HCMV)对脐血多向造血祖细胞(CFU-Mix)体外增殖的影响及黄芪对此的保护作用。方法采用甲基纤维素半固体培养技术,培养、观察、计数HCMV-AD169感染的CFU-Mix集落数、抑制率、集落峰值及集落维持时间;用聚合酶链反应(PCR)和荧光定量PCR技术检测集落细胞内HCMV-AD169DNA。在1∶10感染组中加入黄芪,观察黄芪对HCMV-AD169感染的CFU-Mix集落增殖抑制的干预作用。结果HCMV-AD169感染的CFU-Mix集落数较对照组明显减少(P<0.05),集落数随病毒感染滴度的增高而减少,抑制率随病毒感染滴度的增高而逐渐增加;感染组集落维持时间较对照组明显缩短(P<0.01),但各组集落峰值出现时间无明显差异(P>0.05);用PCR技术在感染组集落细胞内检测到HCMV-AD169DNA;在1∶10感染组中加入黄芪后,CFU-Mix集落数明显提高,集落增殖提高率为38.9%,集落维持时间较1∶10感染组明显延长(P<0.01),荧光定量PCR技术检测集落细胞内HCMV-DNAcopies较1:10感染组明显减少。结论在体外HCMV-AD169对CFU-Mix生长和集落形成有明显的抑制作用,HCMV能直接感染多向造血祖细胞,此可能与临床HCMV感染引起贫血、血小板减少和粒细胞减少有关;黄芪在体外有明显的抗HCMV作用。
Objective To investigate the effects of human cytomegalovirus (HCMV) on the proliferation of cord blood multi-hematopoietic progenitor cells (CFU-Mix) in vitro and the protective effects of Radix Astragali. Methods The number of CFU-Mix colonies, the inhibition rate, the peak of colonies and the colony maintenance time of HCMV-AD169 infection were cultured, observed and counted by using methylcellulose semi-solid culture technique. Polymerase chain reaction (PCR) and fluorescence quantitative PCR The colony cells HCMV-AD169 DNA was detected. Astragalus was added into the 1:10 infection group to observe the intervention effect of Astragalus on the proliferation inhibition of HCMV-AD169 infected CFU-Mix. Results The number of CFU-Mix colonies infected by HCMV-AD169 was significantly lower than that of the control group (P <0.05). The number of colonies decreased with the increase of virus titer, and the inhibition rate increased gradually with the increase of virus titer. (P <0.01), but there was no significant difference in peak time of each group (P> 0.05). HCMV-AD169 DNA was detected in the colonies of infected cells by PCR technique. In 1:10 infected group After adding Astragalus, the number of CFU-Mix colonies increased significantly, the rate of increase of colony proliferation was 38.9%, the colony maintenance time was significantly longer than 1:10 infection group (P <0.01), Fluorescent quantitative PCR detection of colony cells within HCMV-DNAcopies 1:10 infection group was significantly reduced. Conclusions HCMV-AD169 can significantly inhibit the growth and colony formation of CFU-Mix in vitro. HCMV can directly infect multidirectional hematopoietic progenitor cells, which may be related to the clinical HCMV infection causing anemia, thrombocytopenia and neutropenia. Astragalus in vitro There is a clear anti-HCMV effect.