中波紫外线照射对HaCaT细胞Janus激酶-信号转导及转录激活因子信号通路和白细胞介素-10、12表达的影响  

Effects of UVB irradiation on the activity of JAK-STAT signaling pathway and expression of IL-10 and IL-12 in HaCaT cells

作　　者：喻晶 张宜 朱以良 陈芳 刘琴 YU Jing;ZHANG Yi;ZHU Yiiang;CHEN Fang;LIU Qin(Laboratory Animal Room,General Hospital of Central Theater Command,Wuhan 430070,China)

机构地区：[1]中部战区总医院实验动物室,湖北武汉430070

出　　处：《临床皮肤科杂志》2024年第11期641-646,共6页Journal of Clinical Dermatology

基　　金：中部战区总医院育英计划(ZZYFH202117)。

摘　　要：目的:探讨中波紫外线辐照对人永生化表皮细胞(HaCaT)中Janus激酶(JAK)-信号转导及转录激活因子(STAT)信号通路和相关炎症因子白细胞介素(IL)-10、IL-12表达的影响。方法:实验分正常对照组(0 m J/cm^(2))和3个不同剂量辐照组,辐照组分别采用紫外线(30、60、90 mJ/cm^(2))辐照HaCaT细胞,辐照后24 h,通过光镜观察细胞形态的变化;细胞计数法(CCK8)检测细胞活力;实时荧光定量PCR检测IL-10 mRNA和IL-12 mRNA的表达水平;酶联免疫吸附试验检测IL-10和IL-12的含量;蛋白免疫印迹实验检测JAK1、JAK2、p-JAK1、p-JAK2和p-STAT1、p-STAT3的蛋白表达水平。结果:与正常对照组相比,HaCaT细胞经不同剂量紫外线辐照24 h后,细胞出现皱缩、变成圆形、细胞核突出,细胞活力随着辐照剂量的增加而降低(P<0.01);IL-10、IL-12的含量及IL-10 mRNA、IL-12 mRNA的表达水平随着辐照剂量的增加而升高(P<0.05);p-JAK1、p-JAK2和p-STAT1、p-STAT3蛋白表达水平随着辐照剂量的增加而升高(P<0.05);JAK1和JAK2蛋白表达水平的变化差异无统计学意义。结论:中波紫外线辐照HaCaT细胞引起了JAK-STAT信号通路及炎症相关因子IL-10、IL-12的表达改变。Objective:To investigate the role of JAK-STAT signaling pathway and the expression of IL-1O and IL-12 in hu-man immortalized epidermal cells(HaCaT)under UVB irradiation.Methods:HaCaT cells were irradiated with different doses of UVB(0,30,60,90 mJ/cm^(2))for 24 h.Afterward,microscopy was performed to observe cell morphology,cell viability was mea sured by CCK-8,mRNA levels of IL-10 and IL-12 by real-time fluorescent quantitative PCR,concentrations of IL-10 and IL-12 in the culture lysate of HaCaT cells by enzyme linked immunosorbent assay(ELISA),protein levels of JAK1,JAK2 and p-JAK1,p-JAK2,p-STAT1,p-STAT3 by Western-blot.Results:Compared with control group,cell shrinkage and cell viability decreased;the mRNA levels of IL-10 and IL-12 increased(P<0.05),the concentrations of IL-10 and IL-12 increased(P<0.05),and the protein levels of p-JAKI,p-JAK2,p-STAT1,p-STAT3(except JAK1 and JAK2)increased with the dosage of UVB(P<0.05).Conclusion:The JAK-STAT signaling pathway,IL-10 and IL-12 participate in injury of HaCaT cells under UVB irradiation.

关 键 词：中波紫外线 人永生化表皮细胞 JAK-STAT信号通路 IL-10 IL-12 

分 类 号：R751[医药卫生—皮肤病学与性病学]