摘要
目的探究人巨细胞病毒活动性感染对早孕绒毛细胞基质金属蛋白酶9(MMP9)和组织型基质金属蛋白酶抑制剂(TIMP1)活性的影响。方法收集2016年10月-2018年10月在湘潭市妇幼保健院和衡阳市第一人民医院自愿接受流产的健康孕妇的绒毛,人巨细胞病毒感染后,免疫组化检测细胞角蛋白7(CK7)、人波形蛋白(Vim)、HER-2基因(c-erbB-2)的表达以观察绒毛细胞纯度。免疫组化检测pp65抗原用于观察人巨细胞病毒(HCMV)的感染情况。Western Blot、荧光定量PCR(RT PCR)和酶联免疫吸附测定(ELISA)用于检测HCMV感染后早孕绒毛细胞中MMP9和TIMP1的表达水平。结果免疫组化染色显示,分离的原代细胞为高纯度绒毛细胞且HCMV可感染原代早孕绒毛细胞。PCR结果显示,HCMV组MMP9基因表达水平为0.23±0.08,低于对照组(P<0.05),TIMP1基因表达水平为5.49±1.76,高于对照组(P<0.05)。HCMV感染后第4天,HCMV组早孕绒毛细胞胞质中MMP9蛋白表达水平为0.18±0.08,低于对照组(P<0.05),TIMP1蛋白表达水平为0.97±0.16,高于对照组(P<0.05)。此外,与模拟感染细胞相比,感染HCMV的巨噬细胞中MMP-9活性持续降低。感染后第7天HCMV组上清液中MMP9蛋白水平低于对照组(P<0.05);感染后第4天和第7天HCMV组上清液中TIMP1蛋白水平高于对照组(P<0.05)。结论 HCMV感染可显著下调原代早孕绒毛细胞MMP9的基因和蛋白表达,上调TIMP1的基因和蛋白表达。
OBJECTIVE To investigate the effect of human cytomegalovirus active infection on the activity of MMP9 and TIMP1 in villi cells of early pregnancy. METHODS Healthy pregnant women, who volunteered for abortion at the Xiangtan maternal and child health care hospital and the First people’s hospital of Hengyang from October 2016 to October 2018 were recruited and their villi were collected. Villi cells were infected by Human cytomegalovirus(HCMV). Immunohistochemical technique was used to detect the expression of CK7, Vim and c-erbB-2 to observe the purity of villi cells, and Pp65 antigen was used to discriminate cells infected with HCMV from them. Western Blot, fluorescence quantitative PCR and ELISA were used to detect the expression levels of MMP9 and TIMP1 in villi cells of early pregnancy after HCMV infection. RESULTS Immunohistochemical staining showed that the isolated primary cells were high purity of villi cells and HCMV could infect the primary early pregnancy villi cells. Fluorescence quantitative PCR showed the mRNA expression of MMP9 in the HCMV group was 0.23±0.08, which was significantly lower than that in the control group(P<0.05). MRNA expression of TIMP1 was 5.49±1.76, which was significantly higher than that in the control group(P<0.05).Four days after the infection, the protein level of MMP9 in the HCMV group was 0.18±0.08, which was significantly lower than that in the control group(P<0.05), whereas the protein level of TIMP1 was 0.97±0.16, which was significantly higher than that in the control group(P<0.05). Besides, the protein activity of MMP9 in marophage cells infected with HCMV was consistently decreasing. Seven days after infection, the MMP9 protein level in the supernatant of the HCMV group was was significantly lower than that in the control group(P<0.05). On the 4 th day and 7 th day after infection, the TIMP1 protein level in the supernatant of the HCMV group were significantly higher than that in the control group(P<0.05). CONCLUSION HVMV infection could significantly down-regulate the mRNA expression and protein level of MMP9 in the early pregnancy villus cells and up-regulate the gene expression and protein level of TIMP1.
作者
蒋娜
胡蘅芬
詹胜红
陶静
周芳
JIANG Na;HU Heng-fen;ZHAN Sheng-hong;TAO Jing;ZHOU Fang(Hunan Polytechnic of Environment and Biology,Hengyang、Hunan 421005,China;不详)
出处
《中华医院感染学杂志》
CAS
CSCD
北大核心
2020年第6期913-918,共6页
Chinese Journal of Nosocomiology
基金
衡阳市科技计划基金资助项目(S2018F9031018294)。
