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[更昔洛韦与黄芪对体外巨细胞病毒感染后脐血造血祖细胞增殖的影响]

[Influence of ganciclovir and astragalus membranaceus on proliferation of hematopoietic progenitor cells of cord blood after cytomegalovirus infection in vitro].

作者信息

Liu Wen-jun, Liu Bin, Guo Qu-lian, Zhang Ying-chun, Yuan Ya-Jie, Fu Xiao-dong, Deng Zheng-hua, Lin Jiang

机构信息

Department of Pediatrics, Affiliated Hospital, Lu Zhou Medical College, Lu Zhou 646000, China.

出版信息

Zhonghua Er Ke Za Zhi. 2004 Jul;42(7):490-4.

Abstract

OBJECTIVE

Cytomegalovirus (CMV) infection was greatly common in the world. CMV infection produces usually mild or asymptomatic infections in individuals with normal immune responses, whereas it may cause serious disease in immunosuppressive patients. Clinical manifestations include suppression of myelopoiesis, a mononucleosis like syndrome, hepatosplenomegaly, lymphadenopathy, thrombocytopenia, and hemolytic anemia. In patients undergoing bone marrow transplantation CMV remains the most common infectious causes of morbidity and mortality. But the treatment drugs with specific effect for CMV was fewer at the present. This study was to investigate the effect of CMV on proliferation of colony forming unit granulocyte-macrophage (CFU-GM), CFU-erythroid (CFU-E), brust forming unit-erythroid (BFU-E), CFU-multipotential (CFU-Mix) and CFU-megakaryocyte (CFU-Mk) progenitor cells of cord blood (CB) with the presence of ganciclovir (GCV) and astragalus membranaceus in vitro.

METHODS

Twenty CB samples were collected from fetal umbilical vein of normal term spontaneous delivery neonates. The colony forming unit-assay was applied to observe the suppression effect of CMV-AD169 strain on CFU-GM, CFU-E, BFU-E, CFU-Mix and CFU-Mk of CB with the presence of GCV and astragalus membranaceus in vitro. The technique of PCR was used to demonstrate the existence of CMV-AD169 DNA in the colony cells of cultured CFU-GM, CFU-E, BFU-E, CFU-Mix and CFU-Mk.

RESULTS

(1) The numbers of CFU-GM, CFU-E, BFU-E, CFU-Mix and CFU-Mk colonies in CMV infection groups were significantly less than those in blank and mock group, respectively. The last time of colonies in groups with CMV infection was significantly shorten compared with the blank and mock group. (2) CMV-DNA was positively detected in the colony cells of CMV infection groups by PCR, while negative in the control groups. (3) The lasting time of CFU-GM, CFU-E, BFU-E, CFU-Mix and CFU-Mk colonies infected with CMV extended significantly with the presence of astragalus membranaceus and GCV, and the numbers of those increased significantly compared with the CMV infection group, respectively. The increasing rate of colonies was 27.2%, 45.2%, 49.1%, 39.0% and 11.9% with astragalus membranaceus group, 37.4%, 74.2%, 71.7%, 67.4% and 38.9% with GCV group, 53.6%, 83.8%, 88.7%, 87.8% and 61.5% with astragalus membranaceus and GCV group, respectively.

CONCLUSIONS

The differentiation and proliferation of CFU-GM, CFU-E, BFU-E, CFU-Mix and CFU-Mk were significantly inhibited after infected with CMV-AD169 strain. The suppression effect of CMV-AD169 on CFU-GM, CFU-E, BFU-E, CFU-Mix and CFU-Mk was inhibited with the presence of GCV and astragalus membranaceus in vitro. This suggested that CMV-AD169 may be inhibited or killed by GCV and Astragalus Membranaceus in vitro.

摘要

目的

巨细胞病毒(CMV)感染在全球极为常见。CMV感染在免疫反应正常的个体中通常产生轻度或无症状感染,而在免疫抑制患者中可能导致严重疾病。临床表现包括骨髓抑制、单核细胞增多症样综合征、肝脾肿大、淋巴结病、血小板减少和溶血性贫血。在接受骨髓移植的患者中,CMV仍然是发病和死亡的最常见感染原因。但目前针对CMV有特效的治疗药物较少。本研究旨在体外观察在更昔洛韦(GCV)和黄芪存在的情况下,CMV对脐血(CB)中粒-巨噬细胞集落形成单位(CFU-GM)、红系集落形成单位(CFU-E)、爆式红系集落形成单位(BFU-E)、多能集落形成单位(CFU-Mix)和巨核细胞集落形成单位(CFU-Mk)祖细胞增殖的影响。

方法

从足月自然分娩新生儿的胎儿脐静脉采集20份CB样本。应用集落形成单位测定法,在体外观察CMV-AD169株在GCV和黄芪存在的情况下对CB的CFU-GM、CFU-E、BFU-E、CFU-Mix和CFU-Mk的抑制作用。采用PCR技术检测培养的CFU-GM、CFU-E、BFU-E、CFU-Mix和CFU-Mk集落细胞中CMV-AD169 DNA的存在情况。

结果

(1)CMV感染组中CFU-GM、CFU-E、BFU-E、CFU-Mix和CFU-Mk集落数分别显著少于空白组和 mock组。CMV感染组集落的持续时间与空白组和mock组相比显著缩短。(2)通过PCR在CMV感染组的集落细胞中检测到CMV-DNA呈阳性,而对照组为阴性。(3)在黄芪和GCV存在的情况下,感染CMV的CFU-GM、CFU-E、BFU-E、CFU-Mix和CFU-Mk集落的持续时间显著延长,且其数量分别比CMV感染组显著增加。黄芪组集落增加率分别为27.2%、45.2%、49.1%、39.0%和11.9%;GCV组分别为37.4%、74.2%、71.7%、67.4%和38.9%;黄芪和GCV组分别为53.6%、83.8%、88.7%、87.8%和61.5%。

结论

感染CMV-AD169株后,CFU-GM、CFU-E、BFU-E、CFU-Mix和CFU-Mk的分化和增殖受到显著抑制。体外在GCV和黄芪存在的情况下,CMV-AD169对CFU-GM、CFU-E、BFU-E、CFU-Mix和CFU-Mk的抑制作用受到抑制。这表明在体外GCV和黄芪可能抑制或杀灭CMV-AD169。

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