肿瘤坏死因子α和白细胞介素1β在结膜松弛症发病中的炎性机制及意义  

Inflammatory mechanism and significance of tumor necrosis factor-α and interleukin-1β in pathogenesis of conjunctivochalasis

作　　者：柯梅青 张兴儒[1] 周桂贞[1] 马艳春[2] 李青松[1] 项敏泓[1] 

机构地区：[1]上海中医药大学附属普陀医院眼科,200062 [2]上海中医药大学附属普陀医院中心实验室,200062

出　　处：《中华实验眼科杂志》2017年第4期325-331,共7页Chinese Journal Of Experimental Ophthalmology

基　　金：上海市卫生局科研基金项目（20124415）

摘　　要：背景结膜松弛症（CCh）是年龄相关性眼表疾病，研究发现CCh患者结膜组织及泪液中炎性细胞因子表达升高，推测炎性细胞因子可能参与CCh的发病，但其作用机制尚未完全阐明。研究还发现CCh患者结膜组织中基质金属蛋白酶（MMPs）的表达上调，而炎性因子与MMPs在CCh发病中是否有关联尚不清楚。目的探讨肿瘤坏死因子α（TNF-α）和白细胞介素1β（IL-1β）对CCh患者结膜成纤维细胞中MMP-1、MMP-3及TNF-α刺激产生的肿瘤坏死因子刺激基因6（TSG-6）和穿透素3（PTX3）表达的影响。方法采用组织块培养法分别对CCh和白内障患者手术中采集的结膜组织进行培养，将培养的2种结膜成纤维组织各分为3个组，分别用含20ng／ml的PBS、TNF-α或IL-1B的细胞全培养基培养细胞4h，分别采用实时荧光定量PCR法和Western blot法检测细胞中MMP-1、MMP-3、TSG-6和PTX3 mRNA及其蛋白的表达。结果倒置显微镜下可见第2代细胞的形态和大小一致，为长梭形，呈放射状排列，细胞核呈卵圆形，细胞突起相互交联。CCh组和白内障组间用PBS、TNF-α和IL-1β干预后细胞中MMP-1、MMP-3、TSG-6和PTX3mRNA相对表达量的总体比较差异均有统计学意义（MMP-1 mRNA：F分组=2．611，P=0．116；F干预=161．564，P=0．000；MMP-3 mRNA：F分组=5．201，P=0．029；F干预=211．021，P=0．000；TSG-6 mRNA：F分组=47．209，P=0．000；F干预=119．340，P=0．000；PTX3 mRNA：F分组=40．512，P=0．000；F干预=93．935，P=0．000）。与白内障组PBS干预后比较，CCh组PBS干预后MMP-1、MMP-3、TSG-6和PTX3 mRNA的相对表达量均明显升高，差异均有统计学意义（均P〈0．05）；与CCh组PBS干预后比较，CCh组TNF-α和IL-1β干预后细胞中MMP-1、MMP-3、TSG-6和PTX3 mRNA的相对表达量明显升高，差异均有统计学意义（均P〈0．05）；与白内障组PBS干预后比较，白内障组TNF-α和IL-1β干预后细胞中MMP-1Background Conjunctivochalasis （CCh） is a common age-related ocular surface diseases. Researches showed that the inflammatory factors are upregulated in conjunctiva and tear of CCh patients, inferring inflammation participates in the pathogenesis of CCh,however,its mechanism is unclear now. Studies also showed that the expression of matrix metalloproteinases （MMPs） in CCh eonjunctiva is elevated. However,the association between inflammatory factors and MMPs is unknown. Objective This research was to observe the effects of tumor necrosis factor-α （TNF-α） and interleukin-1β （ IL-1β） on the expression of MMP-1, MMP-3 ,tumor necrosis factor-stimulated gene-6（TSG-6） and pentraxin-3 （PTX3） in cultured CCh fibroblasts. Methods Conjunctival fibroblasts were isolated and cultured from CCh-derived and cataract-derived human conjunctiva explants. The cells were treated using 20 ng/ml PBS,TNF-α or IL-1β for 4 hours,respectively. The expression levels of MMP-1 ,MMP-3, TSG-6 and PTX3 genes and proteins were detected by quantitative real time PCR and Western blot assay,respectively. Results The second-generation cells showed a long and fusiform shape with ovoid nucleus and radial agreement, and the cell processors connected each other. The differences of the relative expression levels of MMP-1, MMP-3 ,TSG-6 and PTX3 mRNA in the cells were significantly different between CCh group and cataract group after being treated by PBS,TNF-α and IL-1β （ MMP-1 mRNA ： Fgroup = 2.611, P = 0. 116 ; Fintervene = 161. 564, P = 0. 000 ; MMP-3 mRNA ： Fgroup = 5. 201, P = 0. 029 ; Fintervene= 211. 021, P = 0. 000 ; TSG-6 mRNA ： Fgroup = 47. 209, P = 0. 000 ; Finterene= 119. 340, P = 0. 000 ; PTX3 mRNA ： Fgroop = 40. 512, P = 0. 000 ; Fintervene = 93. 935, P = 0. 000 ）. Compared with the cataract group with PBS treatment,the expression levels of MMP-1, MMP-3,TSG-6 and PTX3 mRNA were significantly elevated in the CCh group with PBS treatment or the cataract group with TNF-α and IL-1β treatment （ all at

关 键 词：结膜松弛症 炎症 炎性细胞因子 基质金属蛋白酶 肿瘤坏死因子刺激基因6 穿透素3 细胞培养 人 

分 类 号：R777.3[医药卫生—眼科]