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妊娠期低分子质量肝素干预对新生鼠肝细胞先天性感染人巨细胞病毒的影响
引用本文:李继安,顾绍庆,叶亮英,郑意,顾小海,张晓鸣,朱广丽. 妊娠期低分子质量肝素干预对新生鼠肝细胞先天性感染人巨细胞病毒的影响[J]. 实用儿科临床杂志, 2011, 26(10): 792-794,797
作者姓名:李继安  顾绍庆  叶亮英  郑意  顾小海  张晓鸣  朱广丽
作者单位:1. 江苏大学临床医学院,江苏,镇江,212001
2. 江苏大学附属人民医院,儿科,江苏,镇江,212001
基金项目:江苏省自然科学基金(BK2008238)
摘    要:目的 探讨妊娠期应用低分子质量肝素(LMWH)对新生鼠肝细胞先天性感染人巨细胞病毒(HCMV)的病理改变和病毒DNA载量的影响.方法 取健康纯系清洁级Balb/c小鼠48只.随机选择配对,每对雌雄小鼠各1只.实验组20对,正常对照组4对.实验组每4对为1组,共5组,每只分别腹腔内注射6.0 log半数组织培养感染剂量(TCID50)HCMV-AD169.正常对照组4对,每只腹腔内注射相应细胞维持液高糖改良Eagles培养液(DMEM)1.0 mL.所有小鼠于接种后配对,隔离喂养,并观察受精情况,记录受精时间.发现妊娠后,分别予实验Ⅰ组皮下注射LMWH 1 000 U·kg-1,实验Ⅱ组皮下注射 LMWH 500 U·kg-1,实验Ⅲ组皮下注射LMWH 250 U·kg-1,实验Ⅳ组肌肉注射更昔洛韦(GCV)60 mg·kg-1,各10 d;实验Ⅴ组为阳性对照组,孕鼠不注射LMWH和GCV.每组随机取10只新生鼠,出生1 d处死,取出肝脏,进行病理学检查和荧光定量PCR检测.结果 实验Ⅰ组肝组织病理损害较阳性对照组明显减轻,HCMV DNA载量[(1.60±1.12)×103拷贝数/50 mg]较阳性对照组[(5.91±2.91)×107拷贝数/50 mg]显著降低(P<0.01),与实验Ⅳ组[(1.75±1.05)×103拷贝数/50 mg]比较差异无统计学意义(P>0.05).实验Ⅱ组肝组织炎性反应较阳性对照组稍减轻.实验Ⅲ组肝组织炎性反应与阳性对照组比较无明显变化,实验Ⅲ组HCMV DNA载量为(5.49±2.76)×107拷贝数/50 mg,与阳性对照组比较差异无统计学意义(P>0.05).结论在妊娠期给予LMWH 1 000 U·kg-1干预能显著降低HCMV AD169株先天性感染Balb/c新生鼠肝细胞病毒感染和复制,减轻细胞炎性反应.

关 键 词:人巨细胞病毒  先天性感染  新生鼠    低分子肝素

Influence of Low Molecular Weight Heparin Interventionin Pregnancy on Congenital Cytomegalovirus Infection in Liver Cells of Newborn Mice
LI Ji-an , GU Shao-qing , YE Liang-ying , ZHENG Yi , GU Xiao-hai , ZHANG Xiao-ming , ZHU Guang-li. Influence of Low Molecular Weight Heparin Interventionin Pregnancy on Congenital Cytomegalovirus Infection in Liver Cells of Newborn Mice[J]. Journal of Applied Clinical Pediatrics, 2011, 26(10): 792-794,797
Authors:LI Ji-an    GU Shao-qing    YE Liang-ying    ZHENG Yi    GU Xiao-hai    ZHANG Xiao-ming    ZHU Guang-li
Affiliation:1(1.Clinical Medical College,Jiangsu University,Zhenjiang 212001,Jiangsu Province,China;2.Depatment of Pediatrics,the People′s Hospital Affiliated to Jiangsu University,Zhenjiang 212001,Jiangsu Province,China)
Abstract:Objective To explore the effects of low molecular heparin(LMWH) intervention in pregnancy on pathological changes in liver tissue and human cytomegelovirus(HCMV) DNA load of newborn mice liver cells which had congenital infection with HCMV.Methods Forty-eight healthy Balb/c mice with pure line and sanitation(half of mice were female) were selected when they were about 10 weeks old and weight 22-30 g.They were randomly paired in male and female.The experimental group had 20 pairs and normal control group had 4 pairs.Every 4 pairs were divided into one experimental group,and 6.0 log tissue culture infective dose 50(TCID50)HCMV-AD169 were taken to intraperitoneally inject in male and female mice.Each of 4 pairs in the normal control group was in intra-abdominal injected corresponding cells maintain fluid dulbecco′s modified Eagles medium(DMEM) 1.0 mL.All mice were matched after in inoculation and isolated feeding,to observe fertilization and recording fertilization time.When pregnancy was confirmed,mice in the experimental group Ⅰ were subcutaneously injected LMWH 1 000 U·kg-1,in experimental group Ⅱ were subcutaneously injected LMWH 500 U·kg-1,in experimental group Ⅲ were subcutaneously injected LMWH 250 U·kg-1 and in experimental group Ⅳ were intramuscularly injected ganciclovir(GCV) 60 mg·kg-1 for 10 days,meanwhile the experimental group Ⅴ for positive control,the pregnant mice were not injected with LMWH or GCV.The newborn mice were killed after production 1 day,randomly taking 10 newborn mice each group,to remove the newborn mice,and its pat-hological examinations were done and fluorescence quantitative-polymerase chain reaction(PCR) detection was performed.Results In experimental group Ⅰ,pathological damage of the liver tissue was much lighter than that in positive control group,its HCMV DNA load was(1.60±1.12)×103 copies/50 mg,which was more significantly lower than that in positive control group [(5.91±2.91)×107 copies / 50 mg](P<0.001);there was no significant difference between experimental group Ⅰ and experimental group Ⅳ [(1.75±1.05)×103 co-pies/50 mg](P>0.05).In experimental group Ⅱ,liver tissue inflammation was sightly relieved compared to the positive control group,but the experimental group Ⅲ,liver tissue inflammation did not change apparently compared to the positive control group,and the HCMV DNA load was(5.49±2.76)×107copies / 50 mg,compared to that in the positive control group,and there was no significant difference(P>0.05).Conclusions After HCMV AD169 strains congenital infection Balb/c newborn mice liver cell,LMWH intervention during pregnancy can significantly lower liver cell virus infection and copy,which results in the relief cell inflammation.
Keywords:human cytomegalovirus  congenital infection  newborn mouse  liver  low molecular weight heparin
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