基于NF-kappaB/p65信号通路探究IL-8调控MCP-1分泌及表达影响晶状体囊袋内残留上皮细胞的迁移  

Effect of interleukin-8 regulation on monocyte chemotactic protein-1 secretion and expression through the NF-kappaB/p65 signaling pathway on the migration of residual epithelial cells in the lens capsule

作　　者：司玮 徐素[1,2] 张宇航 毛一 郭柯宇 胡延忠 张凤妍 Si Wei;Xu Su;Zhang Yuhang;Mao Yi;Guo Keyu;Hu Yanzhong;Zhang Fengyan(Department of Ophthalmology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,Henan Province,China;Zhengzhou University,Zhengzhou 450000,Henen Province,China)

机构地区：[1]郑州大学第一附属医院眼科,河南省郑州市450000 [2]郑州大学,河南省郑州市450000

出　　处：《国际眼科杂志》2025年第4期537-543,共7页International Eye Science

摘　　要：目的:探究炎症因子白介素-8(IL-8)影响后发性白内障(PCO)细胞迁移的过程中对晶状体上皮细胞(LEC)分泌的单核细胞趋化蛋白1(MCP-1)的调控作用。方法:构建大鼠晶状体囊袋模型,采用10%胎牛血清培养撕囊后的晶状体囊袋使LEC迁移至后囊30%-50%面积后,撤去血清,将囊袋分为对照组和15 ng/mL IL-8组,并在不同时间点进行拍照观察囊袋LEC迁移。ELISA法和RT-qPCR检测不同组MCP-1的分泌和信使RNA(mRNA)相对表达水平。免疫荧光检测不同组MCP-1表达水平。将SRA01/04细胞分为对照组、15 ng/mL IL-8组和15 ng/mL IL-8+200μmol/L Bindarit(BND)组,用Transwell法检测不同组迁移细胞数量。将SRA01/04细胞分为阴性对照(NC)组、NC+15 ng/mL IL-8组及15 ng/mL IL-8+p65 siRNA组,ELISA法和RT-qPCR检测不同组MCP-1的分泌和mRNA相对表达水平。结果:体外培养的大鼠晶状体囊袋LEC迁移水平显示,在48、72、96 h时15 ng/mL IL-8组囊袋内细胞迁移明显增加(均P<0.05)。ELISA的结果显示,15 ng/mL IL-8组囊袋和SRA01/04细胞与对照组相比较,在12、24 h分泌的MCP-1的水平升高(均P<0.05);RT-qPCR的结果同样显示15 ng/mL IL-8组SRA01/04细胞在12、24 h分泌的MCP-1 mRNA的相对表达水平增高(均P<0.05)。免疫荧光的结果显示,与对照组相比,24 h时15 ng/mL IL-8组的囊袋上皮细胞MCP-1表达水平升高(P=0.007)。Transwell的检测结果显示,与对照组相比,15 ng/mL IL-8组迁移数量增加(P=0.001);与15 ng/mL IL-8组相比,15 ng/mL IL-8+200μmol/L BND组迁移数量减少(P=0.003)。ELISA和RT-qPCR的结果显示,与NC组相比,NC+15 ng/mL IL-8组在12、24 h时的MCP-1分泌和mRNA的相对表达增加(均P<0.01);与NC+15 ng/mL IL-8组比较,15 ng/mL IL-8+p65 siRNA组在12、24 h时的MCP-1分泌和mRNA的相对表达减少(均P<0.01)。结论:IL-8可促进囊袋内残留上皮细胞的迁移,调控晶状体上皮细胞MCP-1的分泌和表达水平,推测其作用机制与NF-κB/p65信号通路有关。AIM:To investigate the effect of interleukin-8(IL-8)on the regulation of monocyte chemotactic protein-1(MCP-1)secreted by lens epithelial cells(LEC)during cell migration in the development of posterior capsule opacification(PCO).METHODS:A rat lens capsule model was established and cultured in medium supplemented with 10%fetal bovine serum.Upon migration of LEC to 30%-50%of the posterior capsule,serum was removed.The capsule was subsequently divided into two groups:a control group and an IL-8(15 ng/mL)treatment group.LEC migration was captured at multiple time points.The secretion and mRNA expression of MCP-1 were quantified using ELISA and RT-qPCR,respectively.Immunofluorescence was used to assess MCP-1 expression in the different experimental groups.SRA01/04 cells were divided into three groups:control,IL-8(15 ng/mL),and IL-8(15 ng/mL)+200μmol/L Bindarit(BND)groups,with migration measured by the Transwell assay.Additionally,SRA01/04 cells were divided into negative control(NC),NC+15 ng/mL IL-8,and 15 ng/mL IL-8+p65 siRNA groups,and MCP-1 secretion and mRNA expression were further analyzed by ELISA and RT-qPCR.RESULTS:LEC migration in the rat lens capsule cultured in vitro showed that the cells migration of the 15 ng/mL IL-8 group significantly increased at 48,72 and 96 h(all P<0.05).ELISA results revealed that MCP-1 levels in SRA01/04 cells from the 15 ng/mL IL-8-treated group were markedly higher than those in the control group at both 12 and 24 h(all P<0.05).RT-qPCR analysis also demonstrated a significant increase in MCP-1 mRNA expression in the 15 ng/mL IL-8 group at both time points(all P<0.05).Immunofluorescence staining indicated greater MCP-1 expression in capsular epithelial cells of the 15 ng/mL IL-8 group at 24 h(P=0.007).Transwell assays further confirmed increased cell migration in the 15 ng/mL IL-8 group compared to the control group(P=0.001),while the migration reduced in the 15 ng/mL IL-8+200μmol/L BND group compared to the 15 ng/mL IL-8 group(P=0.003).Moreover,ELISA and RT-qPCR results demonst

关 键 词：后发性白内障 白介素-8(IL-8) 晶状体上皮细胞 细胞迁移 单核细胞趋化蛋白1(MCP-1) NF-κB/p65信号通路 

分 类 号：R73[医药卫生—肿瘤]