JAK/STAT途径调节瘦素诱导的肝星状细胞Ⅰ型胶原基因的表达  

JAK/STAT pathway mediates leptin-induced Ⅰ collagen mRNA expression in human hepatic stellate cells

作　　者：牛丽文[1] 曹琦[2] 李俊[2] 杨镇[3] 王晓红[2] 

机构地区：[1]厦门大学附属中山医院普外科,福建厦门361004 [2]安徽医科大学药学院,安徽合肥230032 [3]华中科技大学同济医学院附属同济医院普外科,湖北武汉430030

出　　处：《中国药理学通报》2007年第10期1280-1285,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30470755)

摘　　要：目的研究瘦素对肝星状细胞(HSC)α1(Ⅰ)型胶原mRNA表达和蛋白合成的影响,探讨Janus激酶/信号传导及活化转录因子(JAK/STAT)信号传导通路在瘦素诱导的HSC胶原基因表达过程中的作用。方法采用RT-PCR法、ELISA法和Western blot法分别检测不同浓度的瘦素对人肝星状细胞株LX-2α1(Ⅰ)型胶原mRNA表达、蛋白合成以及JAK1、STAT3磷酸化状态的影响;采用Western blot法和RT-PCR法分别观察JAK1抑制剂AG490对瘦素诱导的JAK1磷酸化和α1(Ⅰ)型胶原mRNA表达的影响;采用Western blot法分别观察AG490、LX-2转染STAT3反义寡核苷酸(STAT3-ASON)对瘦素诱导的STAT3磷酸化状态的影响;应用RT-PCR法检测LX-2转染STAT3-ASON对瘦素诱导的α1(Ⅰ)型胶原mRNA表达的影响。结果瘦素增加LX-2α1(Ⅰ)型胶原mRNA表达和蛋白合成,呈剂量效应关系,当瘦素浓度为80μg·L-1时达到最大值;瘦素促进JAK1、STAT3磷酸化,呈时间效应关系;AG490完全阻断瘦素诱导的JAK1、STAT3磷酸化和α1(Ⅰ)型胶原mRNA的表达;LX-2转染STAT3-ASON阻断瘦素诱导的STAT3磷酸化和α1(Ⅰ)型胶原mRNA的表达。结论瘦素通过增加HSCα1(Ⅰ)型胶原mRNA表达和蛋白合成而在肝纤维化发生发展过程中发挥重要的作用,JAK/STAT信号传导通路参与并调节该过程,AG490和LX-2转染STAT3-ASON可有效阻滞此传导途径。在人HSC中,活化的JAK1和STAT3信号可作为肝纤维化治疗新的分子靶点。Aim To investigate the effects of leptin on α1（Ⅰ ） collagen mRNA expression and protein production, and the roles of Janus kinase/signal transducers and activators transcription （JAK/STAT） signaling transduction pathway in increased α1（Ⅰ ） collagen mRNA expression stimulated by leptin in activated hepatic stellate cells （ HSCs ）. Methods Firstly, α1（Ⅰ ） collagen mRNA expression and protein production as well as JAK1 and STAT3 phosphorylation induced by leptin at different doses in a human HSC cell line, LX-2 were determined by RT-PCR, ELISA, and Western-blot. Secondly, the effects of JAK1 inhibitor AG490 on JAK1 phosphorylation and α1（Ⅰ） collagen mRNA expression stimulated by leptin were detected by Western blot and RT-PCR. Thirdly, the roles of AG490 and transfection with STAT3 antisense oligonucleotide（STAT3-ASON） in STAT3 phosphorylation after leptin were detected by Western blot. Finally, the effect of transfection with STAT3-ASON on α1（Ⅰ） collagen mRNA expression after leptin was measured by RT-PCR. Results Leptin increased α1（Ⅰ） colla gen mRNA expression and protein production in a dose-dependent manner in LX-2, reaching a maximal level at 80 μg · L^-1 leptin. In addition, phosphorylation of JAKI and STAT3 after leptin exhibited a timedependent effect. Besides, JAKI inhibitor AG490 completely blocked JAKI and STAT3 phosphorylation and increased in α1（Ⅰ ） collagen gene expression after leptin in LX-2. Transfection with STAT3-ASON blocked STAT3 phosphorylation and increased α1（Ⅰ） collagen mRNA by leptin in LX-2. Conclusion Leptin had a direct action on liver fibrogenesis by stimulating α1（Ⅰ） collagen mRNA expression and protein production in activated HSC, and JAK/STAT signaling transduction pathway was involved in the process. JAKI inhibitor AG490 and transfection with STAT3ASON blocked the transduction pathway effectively in LX-2. Leptin may be an important factor in the development of hepatic fibrosis. Activated JAKI and STAT3 sig

关 键 词：瘦素 肝星状细胞 JAK/STAT信号传导通路 Ⅰ型 胶原 

分 类 号：R322.47[医药卫生—人体解剖和组织胚胎学] R329.24[医药卫生—基础医学]