I型单纯疱疹病毒感染对人角膜上皮细胞基质金属蛋白酶-2和黏着斑激酶表达的影响  

Matrix metallo proteinase-2 and focal adhesion kinase corneal epithelial cell

作　　者：曹婷[1] 邢怡桥[1] 杨燕宁[1] 梅海峰[1] 

机构地区：[1]武汉大学人民医院眼科中心,430060

出　　处：《中华实验眼科杂志》2013年第11期1050-1054,共5页Chinese Journal Of Experimental Ophthalmology

摘　　要：背景研究表明，I型单纯疱疹病毒（HSV-1）感染可导致角膜上皮细胞（CECs）基质金属蛋白酶（MMPs）表达、分泌和活性改变。黏着斑激酶（FAK）对MMPs的表达、释放和激活起着重要的调节作用。人角膜上皮感染HSV-1后FAK的表达变化鲜有报道。目的探讨HSV-1感染对体外培养的人CECsMMP-2和FAK表达的影响。方法以HSV-1（F株）感染体外培养的人CECs，应用逆转录PCR（RT—PCR）、免疫印迹法、免疫组织化学法分别于感染后2、20、40h检测人CECsMMP-2、FAK及磷酸化黏着斑激酶（p-FAK）的表达情况。结果RT．PCR法检测结果显示，感染细胞的MMP-2mRNA、FAKmRNA较非感染细胞（即加入无病毒无血清培养液的阴性对照组）的表达明显增强，不同时间点间比较差异无统计学意义，组别与时间点间不存在交互作用（MMP-2mRNA：F目目=0．968，P=0．436；F蚴I=47．649，P=0．000；F女Ⅱ#目=0．757，P=0．536．FAKmRNA：F时间=0．658，P=0．631；F组别=35．182，P=0．000；F交互作用=1．386，P=0．137）。Westernblot法检测结果显示，感染后2h时p-FAK、FAK、MMP-2在感染细胞与非感染细胞间差异均无统计学意义（P’0．05），感染后20h、40h感染细胞的蛋白表达较非感染细胞均明显增强（P〈0．05）。免疫组织化学染色检测结果显示，随着感染时间的延长，MMP-2、FAK、p-FAK蛋白染色阳性细胞数目增多。结论在HSV-1感染早期，FAK的激活可刺激MMPs活性增强，从而加速角膜溃疡及坏死性病变的形成。Background Previous studies showed that after herpes simplex virus-1 （HSV-1） infection of the cornea,matrix metallo proteinase-2 （MMP-2） （produced by corneal cells and corneal epithelial cells） plays an important role in the development of HSK. Focal adhesion kinase （FAK） plays an important role on the expression, release and activation of MMPs. This study explored the expressions of MMP-2 and FAK, which induced by HSV-1 infected human corneal epithelial cells. Objective To investigate MMP-2 and FAK expression in HSV-1 infected human corneal epithelial cells. Methods Human corneal epithelial cells were infected with HSV-1 in vitro to establish cell model of viral infection. The expression of MMP-2 and FAK were detected by reverse transcription PCR （RT-PCR） , Western blot,immunohistochemieal method and immunofluoreseence method at 2 hours ,20 hours and 40 hours after infection. Results At 2 hours,20 hours and 40 hours after infection, the expressionis of MMP-2 mRNA and FAK mRNA were significantly increased in comparison with uninfected cells （MMP-2 mRNA：Ftim, = 0. 968,P = 0. 436 ; F oop = 47. 649, P = 0. 000 ; Finteracion= 0. 757, P ： 0. 536. FAK mRNA ： Ftime= 0. 658, P = 0.631 ; Fgroop= 35. 182, P=0. 000; Finteration= 1. 386, P = 0. 137 ）. Western blot assays showed that there were no significant differences in p-FAK,FAK or MMP-2 expressions between infected cells and control cells after 2 hours （P 〉 0.05 ）, but the expression levels of infected cells were significantly increased at 20 hours and 40 hours （both at P 〈0. 05 ）. lmmunohistochemistry results showed that longer infection time was associated with an increased number of ceils staining for MMP-2, FAK and p-FAK. Conclusions At the initial stage of HSV-1 infected, p-FAK plays an important role in the process of virus invading and MMP-2 activation.

关 键 词：基质金属蛋白酶-2 黏着斑激酶 I型单纯疱疹病毒 角膜炎 

分 类 号：R752.11[医药卫生—皮肤病学与性病学]