白细胞介素22诱导HaCaT细胞表达肝素结合表皮生长因子样生长因子的作用机制探讨  

Mechanisms underlying interleukin-22-induced expression of heparin-binding epidermal growth factor-like growth factor in HaCaT cells

作　　者：刘欣欣[1] 罗素菊[1] 郑焱[2] 许文娟[1] 李颖[3] 刘全忠[1] 

机构地区：[1]天津医科大学总医院皮肤科,300052 [2] 西安交通大学第二医院皮肤科 [3]天津医科大学肺癌研究所,300052

出　　处：《中华皮肤科杂志》2015年第3期179-183,共5页Chinese Journal of Dermatology

基　　金：国家自然科学基金（81201231、81371732）

摘　　要：目的 探讨白细胞介素22（IL-22）诱导HaCaT细胞表达肝素结合表皮生长因子样生长因子（HB-EGF）的相关作用机制.方法 用不同浓度的IL-22（12.5、25、50、100 μg/L）干预处理HaCaT细胞,对照组选择等量磷酸盐缓冲液（PBS）处理.24 h后,提取HaCaT细胞总蛋白进行蛋白免疫印迹,检测丝裂原活化蛋白激酶-细胞外信号调节激酶1/2（MAPK-ERK1/2）通路中磷酸化ERK1/2（P-ERK1/2）的表达和转录激活因子途径（JAK/STAT）中磷酸化JAK2（P-JAK2）和磷酸化STAT3 （P-STAT3）的表达.将HaCaT细胞分4组,分别用PBS、IL-22、MAPK-ERK1/2抑制剂PD98059、JAK2/STAT3通路抑制剂AG490与IL-22共同干预HaCaT细胞,24 h后,提取细胞总蛋白和总mRNA,分别用蛋白免疫印迹法和实时定量逆转录（RT）-PCR法检测不同处理组HB-EGF蛋白和mRNA水平的改变.采用SPSS16.0软件进行单因素方差分析检验组间差异,Bonferroni检验进行多重比较.结果 不同浓度的IL-22干预处理后,HaCaT细胞中P-ERK1/2、P-JAK2和P-STAT3蛋白表达均高于对照组（P＜0.05）.HB-EGF蛋白水平（HB-EGF/内参照的灰度比值）在PD98059组和AG490组分别为0.183±0.020和0.199±0.011,与IL-22组（0.924±0.032）相比,差异具有统计学意义（F值分别为37.700、36.400,均P＜0.05）.HB-EGF mRNA水平在PD98059组和AG490组分别为1.034±0.072和0.989±0.038,与IL-22组（1.844±0.135）相比,差异具有统计学意义（F值分别为11.271、13.429,均P＜0.05）.结论 IL-22可以引起HaCaT细胞中MAPK-ERK1/2和JAK2/STAT3这两条信号通路激活.IL-22诱导HaCaT细胞产生HB-EGF蛋白的作用机制可能与MAPK-ERK1/2和JAK2/STAT3这两条信号通路有关.Objective To investigate the mechanisms underlying intedeukin-22 （IL-22）-induced expression of heparin-binding epidermal growth factor-like growth factor （HB-EGF） in HaCaT cells.Methods Some HaCaT cells were divided into several inverention groups treated with IL-22 at concentrations of 12.5,25,50,100 μg/L,respectively and a control group treated with phosphate buffer saline （PBS）.After 24-hour culture,total proteins were extracted from the HaCaT cells,and Western blot was performed to measure the expression of phosphorylated extracellular signalregulated kinase 1/2 （P-ERK1/2） in the mitogen-activated protein kinase （MAPK）-ERK1/2 pathway,as well as phosphorylated-JAK2 （P-JAK2） and phosphorylated-signal transducer and activator of transcription 3 （P-STAT3） in the JAK2/STAT3 pathway.In a blocking experiment,some HaCaT cells were divided into 4 groups to be treated with PBS,IL-22,PD98059 （an inhibitor of MAPK-ERK1/2） combined with IL-22 （PD98059 group）,AG490 （an inhibitor of JAK2/STAT3） combined with IL-22 （AG490 group）,respectively.After 24-hour treatment,total proteins and mRNAs were extracted from the HaCaT cells followed by Western blot and real-time quantitative reverse transcription-PCR for the measurement of protein and mRNA expressions of HB-EGF respectively.Statistical analysis was carried out with the software SPSS 16.0 by one-way analysis of variance （ANOVA） for intergroup comparisons and by Bonferroni＆#39;s test for multiple comparisons.Results After treatment with IL-22 at the above 4 concentrations,the expressions of P-ERK1/2,P-JAK2 and P-STAT3 in HaCaT cells were all increased compared with the control group （all P 〈 0.05）.The protein and mRNA expression levels （expressed as the HB-EGF/β-actin ratio and 2-△△Cr respectively） of HB-EGF were both significantly decreased in the PD98059 group and AG490 group than in the IL-22 group （protein：0.183 ± 0.020 and 0.199 ± 0.011 vs.0.924 ± 0.032,F =37.700,36.400,respectively,both P 〈 0.05;

关 键 词：白细胞介素-22 成纤维细胞生长因子1 角蛋白细胞 丝裂原激活蛋白激酶类 Janus激酶2 STAT3转录因子 银屑病 HaCaT细胞 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]