摘要
目的 研究单纯疱疹病毒Ⅰ型（herpes simplex virus type 1，HSV-1）感染小鼠眼球以后转录因子T—bet在小鼠外周血中的表达，探讨T—bet的表达与单纯疱疹性角膜基质炎之间的关系。方法 将10^6空斑单位（plague forming unit，PFU）·L^-1的HSV-1 Mckrae毒株接种于BAIB／c鼠的角膜上建立单纯疱疹性角膜基质炎（herpetic stromal keratitis,HSK）动物模型，分别于角膜接种病毒后的第1d、3d、7d、10d、14d、21d及28d，用毛细管取小鼠的左眼眼眶静脉窦血1mL，提取淋巴细胞，用半定量逆转录聚合酶链反应（RT-PCR）检测T-bet mRNA的表达水平；在裂隙灯显微镜下观察小鼠角膜的临床变化，组织学检查角膜的病理改变；用ELISA法检测T-bet蛋白在角膜组织中的表达水平。结果 BALB／c鼠的角膜接种HSV-1后的1～5d，角膜擦拭液中均检测出HSV-1复制，表明小鼠感染了单纯疱疹病毒；裂隙灯显微镜观察：HSV-1感染鼠的角膜后，小鼠均患了急性角膜上皮炎，并于感染后1周内痊愈。其中81．7％（49／60）的小鼠自感染病毒后10d起开始出现角膜基质炎改变，表现为灶状角膜基质混浊，局部角膜新生血管化，角膜基质内大量的炎性细胞浸润。角膜基质混浊逐渐进展，在病毒感染后的第14～21d达到高峰。RT-PCR检测的数据显示：未感染HSV-1的对照组小鼠的外周血中不表达T-bet mRNA；HSV-1感染小鼠的早期即可诱导T—bet mRNA在小鼠外周血中的表达，并且表达持续存在；T—bet mRNA表达的高峰时间（10～28d），位于角膜基质炎发生和发展的过程中。ELISA法检测角膜提取物中的T-bet蛋白显示了相似的结果。结论 HSV-1上调T-bet mRNA在小鼠外周血中的表达；T-bet的表达与角膜基质炎的发生和发展密切相关。
Objective To investigate the expression of T-bet in murine ripheral blood following ocular herpes simplex virus type 1 （HSV-1） infection and to discuss its relationship with herpetic stromal keratitis. Methods Corneas of 60 BALB/c mice were infected with 106 PFU（plague forming unit）·L^-1 of HSV-1 McKrae strain, and ultraviolet-inactivited HSV-1 was used in another 60 mice and 5μL, PBS in 20 control mice. The clinieal evaluation of infected eyes were taken under the slit-lamp microscope, and the histological changes of corneas were observed under the optical microscope. The expression of T-bet mRNA in peripheral blood of mice was detected by reverse transcription polymerase chain reaction （RT-PCR） on the 1st, 3rd, 7th, 10th, 14th, 21st and 28th day, respectively, after corneal inoculation with HSV-1. At the same time, T-bet protein in cornea was detected by enzyme-linked immunosorbant assay （ELISA）. Results Epithelial damage was observed and healed within seven days after infection of the cornea with HSV-1 in BALB/c mice. From the 10th day on, 81.7% （49/60） of the mice developed stromal opacities, and opacification peaked on the 14th～21st day after inoculation. Semi-quantitative RT-PCR analyses showed that there wash＇ t expression of T-bet mRNA in peripheral blood of control mice, whereas expression of T-bet mRNA was seen in the early infection period and reached the highest level from 10 to 28 days after inoculation in peripheral blood of mice infected by HSV-1 and ouly faint expression occurred in the lst～3rd day in peripheral blood of mice infected by inactivated HSV-1. ELISA evaluation of T-bet protein from corneal extracts demonstrated a similar result. HSV-1 infection also induced corneal edema and neutrophilic granulocyte and lymphocyte infiltrations on the 28th day,however, no responses mentioned above were seen on the cornea infected with ultraviolet-inactivated HSV-1. Conclusion HSV-1 mediates up-regulation of T-bet expression in murine peripheral blood. Expression level of T-
出处
《眼科新进展》
CAS
2006年第11期818-822,共5页
Recent Advances in Ophthalmology
基金
辽宁省教育厅高等学校科学技术研究项目(编号:2004D163)~~
