Effect of lipoxin receptor agonist on human cytomegalovirus replication and proliferation

doi:10.3969/j.issn.2095-4344.2014.29.001

Abstract

Abstract:

BACKGROUND: The anti-inflammation and protective effects of lipoxin have been verified in several immunity-related disease models. Preliminary studies of our research group have shown that, lipoxin receptor agonist BML-111 has negative regulation effects on the human cytomegalovirus (HCMV)-induced immunological injury. However, the effect of BML-111 on the HCMV replication remains unclear.

OBJECTIVE: To observe the influence of lipoxin receptor agonist BML-111 on HCMV replication and proliferation in THP-1 macrophages and human embryonic lung fibroblasts.

METHODS: THP-1 macrophages were infected by HCMV AD169 strain, and were divided into three groups: mock infection, HCMV infection, HCMV+BML-111. The final concentration of BML-111 was 100 nmol/L. Cells in each group were collected at 0, 1, 2, 4, 12, 36, 48 hours, the mRNA levels of IE86 and pp65 in the THP-1 macrophages were tested by RT-PCR method. Human embryonic lung fibroblasts were infected with HCMV (MOI=0.1), and were divided into two groups: HCMV infection and HCMV+BML-111. The patho-morphous changes of human embryonic lung fibroblasts were observed under light microscope, and the cell number was measured. The infective virus titer changes in human embryonic lung fibroblasts were examined by plaque assay.

RESULTS AND CONCLUSION: After the macrophages were infected by HCMV, compared with the mock infection group, the mRNA levels of IE86 and pp65 in the HCMV group and HCMV+BML-111 group were increased significantly; compared with the HCMV infection group, the mRNA levels of IE86 and pp65 in the HCMV+BML-111 group were increased significantly in the early stage (within 4 hours) after infection, but the pp65 mRNA levels were decreased significantly in the medium and late stages (24-72 hours) after infection. After human embryonic lung fibroblasts were infected by HCMV, the degree of the patho-morphous in the HCMV+BML-111 group reached 100% 2 days earlier than the of HCMV infection group. The infective virus titer reached the peak 2 days earlier than the HCMV infection group, but no significant difference was found between the two groups. BML-111 accelerates the replication of HCMV in the early stage of infection, but inhibits the expression of pp65 gene in the late stage. BML-111 has no impact on the proliferation of the infective HCMV titer in vitro.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

全文链接：

Key words: cytomegalovirus, cytomegalovirus infections, macrophages, lipoxins, virus replication

CLC Number:

R318

Chen Xiao-hong, Shu Sai-nan, Liu Xing-lou, Wang Hui, Zhang Ju, Du Xiao-yi, Li Ge, Fang Feng . Effect of lipoxin receptor agonist on human cytomegalovirus replication and proliferation[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(29): 4593-4598.

Figures/Tables 1

2.1 BML-111对人巨细胞病毒在人THP-1源巨噬细胞中复制的影响与模拟感染组相比，人巨细胞病毒感染组和人巨细胞病毒+BML-111组IE86及pp65 mRNA表达明显升高(P < 0.05)。其中，人巨细胞病毒感染组和人巨细胞病毒+BML-111组IE86 mRNA均在0 h时即显著升高，人巨细胞病毒组1 h达高峰，人巨细胞病毒+BML-111组2 h达高峰，均在4 h后迅速下降。人巨细胞病毒感染组和人巨细胞病毒+BML-111组pp65 mRNA亦均在0 h显著升高，人巨细胞病毒感染组在36 h达高峰，48 h之后缓慢下降，人巨细胞病毒+BML-111组在1 h即达高峰，4 h后迅速下降。与人巨细胞病毒感染组相比，人巨细胞病毒+BML-111组IE86 mRNA高峰延迟，pp65 mRNA高峰提前，两者峰值均升高(P < 0.05)，见图1，2。 2.2 BML-111对人巨细胞病毒感染人胚肺成纤维细胞后致细胞病变的影响人巨细胞病毒感染后，人胚肺成纤维细胞逐渐发生细胞病变效应，具有特征性表现：开始出现细胞肿胀，并且变大、变圆，在感染早期，病变细胞零星分布，或几个相连；在感染晚期，病变细胞则呈葡萄串样，可以见到多核巨细胞，最后病变细胞坏死、脱落。倒置显微镜下每日观察巨细胞病毒感染后人胚肺成纤维细胞病变程度，可见：第3天，人巨细胞病毒感染组尚未出现细胞病变，而人巨细胞病毒+BML-111组出现5%细胞病变；第5天，人巨细胞病毒感染组出现20%细胞病变，而人巨细胞病毒+BML-111组出现40%细胞病变；第8天，人巨细胞病毒感染组出现65%细胞病变，而人巨细胞病毒+BML-111组出现100%细胞病变，并脱落。与人巨细胞病毒感染组相比，人巨细胞病毒+BML-111组细胞先出现病变(P < 0.05)，并先达到100%病变(P < 0.05)，见图3。 2.3 BML-111对人巨细胞病毒在人胚肺成纤维细胞内感染性病毒滴度的影响用蚀斑法测定人胚肺成纤维细胞内感染性人巨细胞病毒滴度，受人巨细胞病毒感染后，两组细胞内扩增的感染性病毒滴度逐日升高，人巨细胞病毒组在第9天达到峰值，平均滴度为1.13×107 PFU，而人巨细胞病毒+BML-111组在第7天达到峰值，平均滴度为0.87×107 PFU。与人巨细胞病毒组相比，人巨细胞病毒+ BML-111组的病毒滴度先达到峰值(P < 0.05)，但两组病毒滴度峰值差异无显著性意义(P > 0.05)，见图4。"

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