白细胞介素1、白细胞介素6及肿瘤坏死因子α对人晶状体上皮细胞移行的作用  

Effects of interleukin-1 ,interleukin-6 and tumor necrosis factor-α on migration of human lens epithelial cells

作　　者：杜敏娟[1] 周健[1] 

机构地区：[1]第四军医大学西京医院眼科全军眼科研究所,西安710032

出　　处：《中华实验眼科杂志》2012年第10期873-876,共4页Chinese Journal Of Experimental Ophthalmology

基　　金：国家自然科学基金项目（30672292）;新世纪优秀人才支撑计划项目（NCET-06-932）;西京医院学科助推计划项目（XJZT09D02）

摘　　要：背景晶状体上皮细胞（LECs）在保持晶状体的透明性方面发挥重要的作用。研究表明炎症反应参与了一些类型白内障的形成过程，且炎性因子对LECs的生物学行为产生影响，但其对LECs移行的影响尚不明确。目的探讨白细胞介素1d（IL-1d）、IL-1B、IL-6、肿瘤坏死因子α（TNF—α）及4种炎性因子混合物对人LECs细胞系HLEB-3迁移的影响。方法HLEB-3细胞用含质量分数0．5％胎牛血清的DMEM培养液进行饥饿培养，将10μg/L的IL—1α、IL—1β、IL-6、TNF—α及这4种炎性因子的混合物分别加入培养液中作用24h后进行细胞划痕实验和Transwell实验。于上述因子处理后即刻和24h在倒置显微镜下观察并用数码相机记录划痕区，在200倍显微镜下计数划痕线内移行的细胞数量。Transwell实验24h后取出小室用质量分数4％多聚甲醛溶液固定15min，草酸铵结晶紫溶液染色2min，于倒置显微镜下观察，选取12：00、3：00、6：00和9：00位以及中心部位5个观察区在200倍显微镜下进行细胞计数，以评价炎性因子对LECs移行的作用。用仅含0．5％胎牛血清的DMEM培养液进行培养的LECs作为空白对照组。结果划痕实验结果表明，各种炎性因子作用后24h，对照组划痕区LECs数量明显低于IL-1β作用组及TNF—α作用组，差异均有统计学意义（P=0．000、0．000），混合因子组划痕区细胞数量均明显高于IL-1β作用组及TNF-α作用组，差异均有统计学意义（P=0．000、0．000）；IL-1α作用组、IL-6作用组与对照组比较，划痕区细胞数量差异均无统计学意义（P=0．600、0．098）。Transwell实验显示，混合因子组作用24h小室膜下层LECs密度最大，与IL-α作用组、TNF—α作用组相比差异均有统计学意义（P=0．000、0．000），且IL-α作用组、TNF—α作用组小室膜下层LECs数量均多于对照组，差异均有统计学意义（P=0.000、0．000）。IL—1α作�Background Lens epithelial cells （LECs）play an important role in maintaining the lens transparency. Inflammatory cytokines have been clarified to participate in the formation of traumatic and after cataracts. However,the influence of inflammatory cytokines on the migration of LECs is still studying. Objective This study was to investigate the effects of interleukin-1 alpha （IL-1α）, interleukin-I beta （IL-113）, IL-6, tumor necrosis factor alpha（TNF-α） and their mixture on the migration of human LECs in vitro. Methods HLE-B3 cells were cultured in DMEM containing 0.5% fetal bovine serum. 10 μg/L of IL-1α,IL-1β,IL-6,TNF-α or the mixture was added in the medium for 24 hours,respectively. Wound-scratch assay and transwell monolayer permeability assay were used to evaluate the effects of different cytokines on LECs migration. The migrating cell numbers at the scratch zone and the transmembrane cell numbers at 12：00,3：00,6：00,9：00 and the center areas on the transwell chamber were calculated under the inverted microscope, in the control group, LECs were cultured in DMEM with 0. 5% fetal bovine serum only. Results Wound-scratch assay revealed that 24 hours after cytokine incubation, the LECs numbers in the scratch zone were significantly less in the control group compared with the IL-1β group and TNF-α group（P = 0. 000,0. 000）, and those in the mixture group were higher than the IL-1β group and TNF-α group （P=0. 000,0. 000）. However,no significant change was found in migrating LECs number between IL-1α group or IL-6 group and the control group （P = 0. 600,0. 098 ）. Transwell assay displayed that after 24-hour culture with cytokines,the largest density of transmembrane LECs was obtained in the mixture group,with a significant difference in comparison with the IL-1 β group or TNF-α group （ P = 0. 000,0. 000 ）. However, the transmembrane LECs were much more in the [L-113 group or TNF-α group compared with the control group （P = 0. 000,0. 000）. Only a few transmembran

关 键 词：炎性因子 晶状体 上皮细胞 细胞移行 

分 类 号：R363[医药卫生—病理学]