Irf3基因缺失导致角膜组织对HSV-1病毒易感性增强

目的研究IRF3缺失对单纯疱疹性角膜炎的发病进程影响以及病理学依据。方法用HSV-l感染对照组和实验组小鼠角膜,同时用培养基(不含病毒)处理Irf3^-/-小鼠角膜作为溶剂对照组。分别用裂隙灯显微镜、HD-OCT、角膜共聚焦技术观察各组角膜的临床变化;采用组织学技术检查角膜病理学改变;用QRT-PCR检测角膜中HSV-1的表达水平;用蛋白免疫印迹法检测炎性指标在各组角膜组织中的表达水平。用流式细胞术、全身症状记录观察小鼠全身感染情况。结果实验组角膜接种HSV-1后第7天开始出现角膜上皮缺损等角膜基质炎早期症状,并且逐渐加重,发病率持续增高。对照组发病率显著较低,且症状比实验组明显减轻。Irf3^-/-溶剂对照组无明显角膜感染变化。QRT-PCR和蛋白免疫印迹法结果显示,实验组HSV-1、IL-9、IL-10、IL-18、TNF-α表达水平均比对照组、Irf3^-/-溶剂对照组显著较高。脾脏流式细胞术、全身症状记录结果显示实验组比对照组、Irf3^-/-溶剂对照组全身炎性反应明显较重,提示有全身感染。结论 IRF3是角膜HSV-1感染的重要感受信号分子,它的缺失使小鼠发生单纯疱疹病毒性角膜炎的易感性明显增高。

Irf3 gene deletion increases the susceptibility of cornea to HSV-1 infection

To investigate the effect of Inf3 deletion on the pathogenesis of herpes simplex keratitis and the pathological process, HSV-1 was used to infect the cornea of mice in control and experimental groups, while Irf3-/-mouse cornea was treated with culture medium(without virus) as vehicle control group. Then, the clinical changes of cornea were observed by slit lamp microscopy, HD-OCT and corneal confocal technique;the pathological changes of cornea were examined by histological techniques;the expression levels of HSV-1 and inflammatory markers in corneal tissues were detected by QRT-PCR and Western blotting, respectively. Flow cytometry and record of systemic symptoms were used to observe systemic infection of mice. Data showed that the early symptoms of corneal stromal inflammation, such as defect of corneal epithelium, began to appear on the 7 th day after HSV-1 infection of cornea in experimental group, which were gradually aggravated, and the incidence continued to increase. The morbidity of the control group was significantly low and the symptoms were significantly milder than those in the experimental group. Irf3-/-vehicle control group demonstrated no obvious infection of corneal. The results of QRTPCR and Western blot showed that the expression levels of HSV-1, IL-9, IL-10, IL-18 and TNF-α in the experimental group were significantly higher than those in the control group and Irf3-/-vehicle control group.The results of flow cytometry and systemic symptoms record showed that the systemic inflammatory reaction in the experimental group was more severe than that in the control group and Irf3-/-vehicle control group, which indicated systemic infection. This study proves that IRF3 is an important sensory signal molecule of HSV-1 infection in cornea and Irf3 deletion significantly increases the susceptibility of mice to herpes simplex keratitis.