摘要
【摘要】目的观察HCMV感染THP-1源性巨噬细胞后，促炎细胞因子IL-1β表达的时序性变化。方法构建HCMV感染THP-1源性巨噬细胞模型，设立HCMVAD169标准毒株感染组、模拟感染对照组、LPS＋ATP对照组和poly（dA：dT）对照组。用ELISA法测定THP-1源性巨噬细胞培养上清IL-1β水平在病毒感染细胞后1h、3h、6h、12h、24h和48h的时序性变化。分别用real-timePCR和westernblot检测感染后6hIL-1β基因和蛋白的表达水平。结果HCMV感染THP-1源性巨噬细胞6h后，促炎细胞因子IL-1β基因的相对表达量是模拟感染组的7．77倍。HCMV感染THP-1源性巨噬细胞1h后，细胞上清IL-1β表达量逐渐显著增高，感染后3h和6h继续升高，感染后12h达到高峰，一直持续到48h。HCMV感染THP-1源性巨噬细胞6hHCMV感染组IL-1β蛋白表达明显高于模拟感染对照组和poly（dA：dT）对照组，而与LPS＋ATP对照组比较无统计学差异。结论HCMV感染THP-1源性巨噬细胞可诱导IL-1β高表达，且呈时序性增高趋势。
Objective To observe the changes in the expression of pro-inflammatory cytokines IL-1β over time in THP-1 derived macrophages after human cytomegalovirus （HCMV） infection. Methods The model of HCMV infection in THP-1 derived macrophages was constructed, then HCMV infected group, the mock-infected control group, LPS ＋ ATP group and poly （dA ： dT） control group were established. ELISA analysis was performed to measure the levels of IL- 1β in culture supernatant of THP- 1 derived macrophages at 1 hour, 3 hours, 6 hours, 12 hours, 24 hours and 48 hours after infected with HCMV. Real-time PCR and western blot were performed to measure the expression levels of IL-1β gene and protein, respectively. Results The gene expression of pro-inflammatory cytokines IL-1β in HCMV infected group is 7.77 times to the mock-infected control group at 6 hours after HCMV infected THP- 1 derived macrophages. IL-1β in supernatant of THP-l-derived macrophages increased significantly at 1 hour after HCMV infection, was continuously rising at 3 hours and 6 hours subsequently, and reached at peak at 12 hours and maintained till 48 hours. The protein expression of IL-1β in HCMV infected group was significantly higher than the mock-infected control group and poly （dA ： dT） control group at 6 hours after HCMV infected THP-I derived macrophages, while no significant difference between the LPS ＋ ATP control group. Conclusions The expression of IL-1β is induced by HCMV infected THP-1 derived macrophages, and continue to increase over time.
作者
刘兴楼
马迪
舒赛男
刘玲玲
方峰
Liu Xinglou Ma Di Shu Sainan Liu Lingling Fang Feng(Department of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science Technology, Wuhan 430030, Chin)
出处
《国际病毒学杂志》
2017年第3期151-155,共5页
International Journal of Virology
基金
国家自然科学基金项目（81301425、81271807）
