大承气汤含药血清对内毒素刺激的人支气管上皮细胞表达CAV-1、eNOS及NF-κB的影响

Effects of Dachengqi Decoction Containing Serum on the Expressions of Caveolin-1,eNOS,and NF-κB in Lipopolysaccharide Stimulated Human Bronchial Epithelial Cells

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DOI：

中文关键词: 大承气汤内毒素人支气管上皮细胞小凹蛋白-1 内皮型一氧化氮合酶核因子-kB

英文关键词:Dachengqi Decoction lipopolysaccharide human bronchial epithelial cell caveolin-1 endothelial nitric oxide synthase nuclear transcription factor-kappa B

基金项目:国家自然科学基金面上项目(No.30873225);湖北省卫生厅资助项目(No.2010Z-Y20)

作者	单位
杨胜兰	华中科技大学同济医学院附属协和医院中西医结合科
金阳	华中科技大学同济医学院附属协和医院中西医结合科
沈霖	华中科技大学同济医学院附属协和医院中西医结合科
刘建国	华中科技大学同济医学院附属协和医院中西医结合科
樊琼	华中科技大学同济医学院附属协和医院中西医结合科
薛卡明	华中科技大学同济医学院附属协和医院中西医结合科
黄璐	华中科技大学同济医学院附属协和医院中西医结合科
吴嫣然	华中科技大学同济医学院附属协和医院中西医结合科

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中文摘要:

目的观察大承气汤含药血清对内毒素(lipopolysaccharide,LPS)刺激的人支气管上皮细胞(human bronchial epithelial cells,HBECs)表达小凹蛋白-1(caveolin-1、CAV-1),内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)及核转录因子κB(nuclear transcription factor-kappa B,NF-κB)的影响。方法制备大承气汤及大承气汤含药血清。将体外培养的HBECs分为7组进行处理(正常血清组,单纯LPS干预组,低剂量大承气汤含药血清组,中剂量大承气汤含药血清组,高剂量大承气汤含药血清组,西药对照组,空白血清对照组),采用四甲基偶氮唑盐(MTT)比色法、Real-timePCR、免疫细胞化学及Westernblot法检测不同剂量的大承气汤含药血清对内毒素刺激HBECs表达CAV-1、eNOS及NF-κBmRNA水平及蛋白的影响。结果正常血清组HBECs有基础量的CAV-1、eNOS及NF-κBmRNA及蛋白的表达,经LPS刺激后,单纯干预组CAV-1、eNOS及NF-κBmRNA及蛋白表达较正常血清组显著增加(P<0.01),而不同剂量大承气汤含药血清均可抑制CAV-1、eNOS及NF-κBmRNA及蛋白表达。结论大承气汤含药血清能抑制LPS刺激的HBECs中CAV-1、eNOS及NF-κB的表达。

英文摘要:

Objective To observe the effects of Dachengqi Decoction(DD) containing serum on the expressions of caveolin-1(CAV-1),endothelial nitric oxide synthase(eNOS),and nuclear transcription factor-kappa B(NF-κB) in lipopolysaccharide(LPS) stimulated human bronchial epithelial cells(HBECs).Methods The DD and the DD containing serum were prepared.The in vitro cultured HBECs were randomly divided into 7 groups,i.e.,the normal serum control group,the LPS intervention group,the low dose DD serum containing group,the middle dose DD serum containing group,the high dose DD serum containing group,the Western medicine control group,the vehicle serum control group.The effects of DD containing serum at different doses on the mRNA and protein expressions of CAV-1,eNOS,and NF-κB were detected using methyl thiazolyl tetrazolium(MTT) colorimetry,Real-time PCR,immuocytochemical assay,and Western blot.Results The mRNA and protein expressions of CAV-1,eNOS,and NF-κB at the basic levels were detected in the HBECs of the normal serum control group.After stimulated by LPS,the mRNA and protein expressions of CAV-1,eNOS,and NF-κB increased more significantly in the LPS intervention group than in the normal serum control group(P<0.01),while DD containing serums at different doses all could suppress the mRNA and protein expressions of CAV-1,eNOS,and NF-κB.Conclusion DD containing serum could inhibit the expressions of CAV-1,eNOS,and NF-κB in LPS stimulated HBECs.

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