摘要
目的:研究前列腺素E2(PGE2)对转化生长因子-β1(TGF-β1)诱导人胚肺成纤维细胞(HLF)转分化及促胶原(COL)合成作用的干预。方法:HLF细胞生长至融合后分为①对照组、②TGF-β1组、③TGF-β1+PGE2组、④TGF-β1+吲哚美辛组,给予干预24h后用细胞免疫荧光及Westernblotting法观察发生转分化为肌成纤维细胞的标志蛋白α-平滑肌动蛋白(α-SMA)的表达,用RT-PCR法检测结缔组织生长因子(CTGF)mRNA和Ⅰ型胶原(COLⅠ)mRNA的水平,用免疫细胞化学法检测CTGF蛋白表达,比色法检测培养上清中羟脯氨酸含量。结果:TGF-β1可将HLF转分化为α-SMA阳性表达的肌成纤维细胞,PGE2则能抑制这种转分化作用;PGE2可显著减低TGF-β1致肌成纤维细胞、CTGFmRNA与蛋白水平(P<0.05)和COLⅠmRNA水平升高(P<0.05);吲哚美辛升高TGF-β1致HLF肌成纤维细胞CTGF与COLⅠ的表达;放线菌酮预处理肌成纤维细胞3h后再加入TGF-β1+PGE2,其COLⅠmRNA水平明显低于TGF-β1处理组水平(P<0.05)。结论:PGE2可抑制TGF-β1对成纤维细胞的转分化及促胶原合成作用,且可通过依赖CTGF和非依赖2种方式下调COLⅠ的转录。
AIM： To observe the effects of prostaglandin E2 on transforming growth factor-β1（TGF-β1）triggered human lung fetus fibroblast （HLF） transdifferentiation and connective tissue growth factor （ CTGF）, collagen type I （COL I ） expression. METHODS： HLFs were treated with TGF - β1, the cells underwent phenotypic change to myofi- broblast. The marker of myofibroblast - α - smooth muscle actin （α - SMA） was detected by immunofluorescence. The α- SMA content was measured by Western blotting. The changes in CTGF and COL I at transcription levels were estimated by RT - PCR method. CTGF protein expression was evaluated by immunocytochemical. Cell culture medium hydroxyproline amount was measured by colormetric assay. RESULTS： PGE2 blocked TGF-β1 induced α -SMA positive myofibroblast transformation （P〈0.05）. TGF-β1 induced CTGF mRNA expression. COL I mRNA level was decreased when co - in- cubated with PGE2 （ P 〈 0.05 ）. Cycloheximide obviously elevated CTGF and COL I expression in myofibroblast. The COL I mRNA level in myofibroblast pretreated with cycloheximide was decreased when incubated with TGF -β1 and PGE2 compared with TGF -β1 treated group. CONCLUSION： PGE2 inhibits TGF - β1 induced transdifferentiation and collagen production, and reduces COL I expression through CTGF dependent and independent manners.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2008年第5期925-930,共6页
Chinese Journal of Pathophysiology
