摘要
目的:探讨核转录因子激活蛋白-1(activator protein-1,AP-1)在转化生长因子β1(transfor-ming growth factor-β1,TGF-β1)诱导人肺成纤维细胞Ⅰ型胶原分泌中的作用。方法:以人肺成纤维细胞HLF-02细胞系为研究对象,给予10μg/L的TGF-β1刺激,于不同时间点收集细胞,采用RT-PCR和Wester blot检测细胞Ⅰ型胶原转录和分泌;阻断实验中选用AP-1抑制剂姜黄素为阻断剂,凝胶阻滞实验(electrophoretic mobility shift assay,EMSA)检测细胞内AP-1的DNA结合活性变化,同时Western印迹检测Ⅰ型胶原分泌变化。结果:TGF-β1能诱导HLF-02细胞Ⅰ型胶原mRNA的转录和分泌(P<0.05);TGF-β1能提高HLF-02细胞AP-1的DNA结合活力(P<0.05);姜黄素能明显抑制TGF-β1诱导的HLF-02细胞AP-1的DNA结合活力,抑制率分别为17.1%,17.6%,24.2%,31.3%(P<0.05);同时,姜黄素能明显抑制TGF-β1诱导的HLF-02细胞Ⅰ型胶原的分泌,抑制率分别为62.1%,58.8%,62.1%,59.6%(P<0.05)。结论:核转录因子AP-1参与TGF-β1刺激的HLF-02细胞Ⅰ型胶原的分泌调控。
Objective To investigate the role of AP-1 in the secretion of Type I collagen in TGF-β1-stimulated human lung fibroblasts. Methods Human lung fibroblasts cell line （HLF-02） was cultured, and then stimulated with 10μg/L TGF-β1 at different time points. Curcumin was added into the culture medium to inhibit the AP-1 activity before incubating with TGF-β1. AP-1 DNA binding activity was assayed by electrophoretic mobility shift assay （EMSA） , and the expression of Type I collagen was detected by Western blot and RT-PCR. Results TGF-β1 could induce the transcription and secretion of Type I collagen in HLF-02 cells （ P 〈 0.05 ）. TGF-β1 could upregulate the AP-1 DNA binding activity （ P 〈0. 05）. Curcumin （ 5, 10, 15, and 20 μmol/L） could inhibit the AP-1 DNA binding activity in TGF-βl-stimulated cells （the inhibition ratio was 17. 1%, 17.6%, 24.2%, and 31.3% ; P〈0.05）. Curcumin （5, 10, 15, and 20 μmol/L） could also inhibit the secretion of Type I collagen significantly （ the inhibition ratio was 62. 1% ,58.8%, 62.1%, and59.6% ; P〈0.05）. Conclusion AP-1 is responsible for the secrection of TGF-β1 -induced Type I collagen in human lung fibroblasts.
出处
《中南大学学报（医学版）》
CAS
CSCD
北大核心
2007年第5期776-781,共6页
Journal of Central South University ：Medical Science
基金
国家自然科学基金项目(30170399)
中南大学理学发展基金项目(07SDF05)~~
