香烟提取物和脂多糖对人胚肺成纤维细胞TGF-β_1 mRNA及蛋白表达的影响  

Effects of cigarrete smoke extract and lipopolysaccharid e on the expression of transforming growth factor-β_ 1 mRNA and protein in human embryonic lung fibroblasts

作　　者：葛晓娜[1] 熊密[1] 黄召[2] 

机构地区：[1]华中科技大学同济医学院病理学系 [2]华中科技大学同济医学院附属协和医院,湖北武汉430030

出　　处：《中国病理生理杂志》2003年第10期1345-1348,T003,共5页Chinese Journal of Pathophysiology

基　　金：国家"九五"科技攻关项目 (No .96 - 90 6 - 0 2 -16 )

摘　　要：目的 :探讨香烟提取物 (CSE)和脂多糖 (LPS)对体外培养的人胚肺成纤维细胞 (HELF)转化生长因子- β1(TGF - β1)mRNA及其蛋白表达的影响。方法 :应用不同浓度的CSE(1∶5 0、1∶2 5和 1∶10 )、LPS(0 1mg/L、1mg/L和 10mg/L)及CSE(1∶2 5 )与LPS(1mg/L)联合作用于HELF ,37℃作用 2 4h后 ,提取细胞总RNA ,应用逆转录 -多聚酶链反应 (RT -PCR)及免疫细胞化学技术检测TGF - β1mRNA和蛋白表达的变化。结果 :CSE低浓度 (1∶5 0和 1∶2 5 )时可增加HELFTGF - β1mRNA及蛋白的表达 (P <0 0 5 ) ,高浓度 (1∶10 )时未引起TGF - β1mRNA及蛋白表达增强 (P >0 0 5 )。不同浓度的LPS均引起HELFTGF - β1mRNA及蛋白表达增强 (P <0 0 5 )。CSE与LPS联合作用也可增加HELFTGF - β1mRNA及蛋白的表达 (P <0 0 5 )。结论 :一定浓度的CSE和LPS可上调肺成纤维细胞TGF - β1mRNA及蛋白表达。AIM: To examine the effects of cigarette smo ke extract (CSE) and lipo polysaccharide (LPS) on the production of transforming growth factor-β 1 (TGF- β 1 ) mRNA and protein in cultured human embryonic lung fibroblasts (HELF). METHODS: The cultured HELF were incubated with CSE, LPS or CSE in com bination with LP S for 24 hours at 37℃, respectively. The total RNA was extracted from the cells . The expression levels of TGF-β 1 mRNA were evaluated by reverse transcription - p olymerase chain reaction (RT-PCR). The mRNA levels of TGF-β 1 were corrected by GAPDH transcripts, and TGF-β 1 protein levels were determined by immunocytoch emistry. RESULTS: CSE stimulated the TGF-β 1 mRNA expression i n HELF at lower concentr ati ons (1∶50 and 1∶25)( P< 0.05) .TGF-β 1 mRNA production wasn't increased by CSE at higher concentration(1∶10)( P> 0.05). LPS enhanced TGF-β 1 mRNA levels in HELF at a ll three doses (0.1 mg/L, 1 mg/L, and 10 mg/L)( P< 0.05). Increased expressi on of TGF -β 1 mRNA was also observed in HELF treated with CSE(1∶25) in combination wit h LPS( 1 mg/L) ( P< 0.05). Immunocytochemistry demonstrated the results of RT-PCR( P< 0.05). CONCLUSION: CSE and LPS upregulated the expr ession levels of TGF-β 1 mRNA and protein in HELF.

关 键 词：香烟提取物 脂多糖类 人胚肺成纤维细胞 转化生长因子Β mRNA 

分 类 号：R563[医药卫生—呼吸系统] R99[医药卫生—内科学]