CAJ | 학술논문

背景：脂氧素的抗炎及保护效应已经在多种免疫相关疾病模型中被验证，课题组前期研究结果亦显示，脂氧素受体激动剂BML-111对巨细胞病毒诱导的免疫损伤有负调节作用，那么，它对病毒的复制将产生怎样的影响呢？ 　　目的：观察脂氧素受体激动剂BML-111对人巨细胞病毒在THP-1源巨噬细胞及人胚肺成纤维细胞中复制增殖的影响。 　　方法：用HCMV AD169毒株感染THP-1源巨噬细胞(MOI=0.5)，设立模拟感染组、人巨细胞病毒感染组及人巨细胞病毒+BML-111组，BML-111的终浓度为100 nmol/L；于感染后0，1，2，4，12，24，36，48 h收集各组细胞，用RT-PCR法检测细胞中IE86和pp65 mRNA的水平；人巨细胞病毒感染人胚肺成纤维细胞(MOI=0.1)，设立人巨细胞病毒感染组与人巨细胞病毒+BML-111组，逐日光镜观察人胚肺成纤维细胞病变并计数其比率；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。 　　结果与结论：人巨细胞病毒感染巨噬细胞后，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 mRNA 及 pp65 mRNA 表达明显升高；与人巨细胞病毒感染组相比，人巨细胞病毒+BML-111组 IE86 mRNA 及 pp65 mRNA 在细胞感染早期(4 h 内)表达明显升高，但在感染中晚期(24-72 h)，pp65 mRNA的表达明显降低。人巨细胞病毒感染人胚肺成纤维细胞后，与人巨细胞病毒感染组相比，人巨细胞病毒+BML-111组细胞提前2 d达到100%细胞病变；感染性病毒滴度提前2 d达到峰值，但两组病毒滴度峰值差异无显著性意义。结果显示 BML-111在感染早期加快人巨细胞病毒在细胞内的复制速度，在感染晚期抑制pp65基因表达；BML-111在体外对人巨细胞病毒的感染性病毒增殖量无影响。
배경：지양소적항염급보호효응이경재다충면역상관질병모형중피험증，과제조전기연구결과역현시，지양소수체격동제BML-111대거세포병독유도적면역손상유부조절작용，나요，타대병독적복제장산생즘양적영향니？ 　　목적：관찰지양소수체격동제BML-111대인거세포병독재THP-1원거서세포급인배폐성섬유세포중복제증식적영향。 　　방법：용HCMV AD169독주감염THP-1원거서세포(MOI=0.5)，설립모의감염조、인거세포병독감염조급인거세포병독+BML-111조，BML-111적종농도위100 nmol/L；우감염후0，1，2，4，12，24，36，48 h수집각조세포，용RT-PCR법검측세포중IE86화pp65 mRNA적수평；인거세포병독감염인배폐성섬유세포(MOI=0.1)，설립인거세포병독감염조여인거세포병독+BML-111조，축일광경관찰인배폐성섬유세포병변병계수기비솔；식반법측정인배폐성섬유세포내감염성인거세포병독적도。 　　결과여결론：인거세포병독감염거서세포후，여모의감염조상비，인거세포병독감염조화인거세포병독+BML-111조 IE86 mRNA 급 pp65 mRNA 표체명현승고；여인거세포병독감염조상비，인거세포병독+BML-111조 IE86 mRNA 급 pp65 mRNA 재세포감염조기(4 h 내)표체명현승고，단재감염중만기(24-72 h)，pp65 mRNA적표체명현강저。인거세포병독감염인배폐성섬유세포후，여인거세포병독감염조상비，인거세포병독+BML-111조세포제전2 d체도100%세포병변；감염성병독적도제전2 d체도봉치，단량조병독적도봉치차이무현저성의의。결과현시 BML-111재감염조기가쾌인거세포병독재세포내적복제속도，재감염만기억제pp65기인표체；BML-111재체외대인거세포병독적감염성병독증식량무영향。
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 col ected 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 cellnumber 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.