EGR1对晶状体上皮细胞炎症反应及EMT的早期调控作用

Early Regulation of the Early Growth Response Gene-1 on Lens Epithelial Cell Inflammation and Epithelial-Mesenchymal Transition after Cataract Surgery in Mice

目的:探究早期生长反应基因1(EGR1)对小鼠白内障术后晶状体上皮细胞(LECs)炎症反应以及上皮-间质转化(EMT)的早期调控作用。方法:实验研究。选取10~16周龄EGR1敲除小鼠(EGR1^-/-)和野生型小鼠(WT)行白内障手术造模,于造模后0 h,3 h,24 h,48 h,72 h,4 d和5 d处死相应小鼠并取眼球做冰冻切片。免疫荧光染色观察晶状体囊袋中EGR1蛋白、CD11b、LCN2、CXCL1和αSMA的表达情况,Image J定量荧光强度。EGR1^-/-小鼠和WT小鼠组间比较采用配对t检验,组内不同时间点比较采用单因素方差分析。结果:WT小鼠白内障术后LECs中EGR1蛋白表达上调、炎症反应指标CD11b、LCN2、CXCL1表达升高。EGR1^-/-小鼠白内障术后LECs中EGR1蛋白表达无变化,CD11b、LCN2、CXCL1表达量明显小于WT小鼠(均P<0.01)。此外,EGR1^-/-小鼠白内障术后EMT特征性标记物αSMA的表达量、囊袋内有形细胞数量亦明显小于WT小鼠(72 h、4 d和5 d,均P<0.05)。结论:白内障手术可早期激活LECs中的EGR1,激活的EGR1参与调控LECs的炎症反应和EMT,敲除EGR1可以减轻上述炎症反应和EMT。

Objective:To study the early growth response gene-1(EGR1)on the inflammatory response of lens epithelial cells(LECs)and epithelial-mesenchymal transition(EMT)after cataract surgery in mice.Methods:In this experimental study,10-16 week-old EGR1 knockout mice(EGR1^-/-)and wild-type mice(WT)were chosen for cataract surgery,and were sacrificed at 0 h,3 h,24 h,48 h,72 h,4 d,and 5 d after modeling.The eyeballs were harvested for frozen section.Immunofluorescence staining was used to observe the expression of the EGR1 protein,CD11b,LCN2,CXCL1 andαSMA in the lens capsule and the fluorescence intensity was quantified by Image J.A paired t-test and one-way analysis of variance were used for comparison between groups and within groups.Results:The expression of the EGR1 protein in LECs after modeling in WT mice was up-regulated,as well as the expression of the inflammatory markers(CD11b,LCN2 and CXCL1).No difference in the level of EGR1 protein was observed in EGR1^-/-mice after modeling,and the levels of CD11b,LCN2 and CXCL1 in the EGR1^-/-mice were significantly lower than those in the WT mice(all P<0.01).In addition,the expression ofαSMA(a marker of EMT)and the number of tangible cells in the lens capsule of EGR1^-/-mice were also significantly less than those of WT mice(72 h,4 d and 5 d after modeling,all P<0.05).Conclusions:Cataract surgery canactivate EGR1 in LECs at an early stage after surgery.Activated EGR1 participates in regulating the inflammatory response and the EMT of LECs.Knocking out EGR1 can reduce the above-mentioned inflammatory response and EMT.