肺损伤机制研究:脂多糖结合蛋白抑制肽对脂多糖诱导的人单核细胞株核转录因子κB活性的影响  

Mechanism of pulmonary injury: effects of lipopolysaccharide binding protein inhibitory peptide on nuclear factor kappa B activity of human monocyte-like cell line induced by lipopolysaccharide

作　　者：吴学玲[1] 钱桂生[1] 徐德斌[1] 侯一峰[1] 陈维中[1] 

机构地区：[1]解放军第三军医大学新桥医院全军呼吸病研究所,重庆市400037

出　　处：《中国临床康复》2005年第11期89-91,共3页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然科学基金资助项目(30170366)

摘　　要：目的:探讨肺损伤机制和脂多糖结合蛋白(lipopolysaccharide-bindingprotein,LBP)抑制肽对脂多糖诱导的人单核细胞株U937细胞核转录因子κB(nuclearfactorkappaB,NF-κB)活性的影响,研究LBP抑制肽对脂多糖所致肺损伤的作用。方法:佛波脂诱导U937成熟后分为5组。即正常对照组;脂多糖组(10μg/L脂多糖+100μg/L重组人LBP);低剂量抑制肽组;中剂量抑制肽组;高剂量抑制肽组(后3组分别给予10,100和1000μg/L抑制肽)。用免疫细胞化学染色图像分析法和蛋白质定量分析法(Westernblot)测定U937细胞NFκB的活性;ELISA测定培养上清液中肿瘤坏死因子α(TNF-α)水平。结果:脂多糖刺激后NFκBP65进入核内,抑制肽组核易位明显减少。低、中、高剂量抑制肽组P65核浆灰度比分别为0.59±0.06,1.02±0.12和1.08±0.10,明显低于脂多糖组1.34±0.12)(t=4.756,P(<0.01;t=2.235,2.308,P<0.05)。低、中、高剂量抑制肽组TNF-α水平分别为(93.66±2.30),(75.78±6.55)和(71.50±7.75)pg/L,明显低于脂多糖组犤(128.67±39.67)pg/L犦(t=2.216,P<0.05;t=2.423,2.389,P<0.01)。结论:LBP抑制肽抑制了U937细胞NFκB的活性和TNF-α的释放;LBP抑制肽对脂多糖所致肺损伤可能具有潜在的预防作用。AIM:To investigate the mechanism of pulmonary injury and influence of lipopolysaccharide binding protein(LBP) inhibitory peptide on nuclear factor kappa B(NFκ B) activity of human like cell line(U937) induced by lipopolysaccharide(LPS),and study the role of LBP inhibitory peptide in LPS induced pulmonary injury. METHODS:U937 was stimulated by phorbol myristate acetate(PMA 10 μ g/L) and divided into five groups: normal control group, LPS group(10 μ g/L LPS+ 100 μ g/L recombined human LBP),low,middle and high dose inhibitory peptide group(10,100 and 1 000 μ g/L inhibitory peptide respectively).The activity of NFκ B of U937 was evaluated by immunochemical staining and image analysis and Western blot.The level of tumor necrosis factor(TNF α ) in culture supernate was detected with enzyme linked immunoadsordent assay(ELISA). RESULTS:NFκ B P65 subunit entered nuclear after U937 was stimulated by LPS,and the translocation of nuclear in the inhibitory peptide groups was obviously decreased.The gray values of P65 subunit in nuclei and cytoplasm of the low,middle and high dose inhibitory peptide groups(0.59± 0.06,1.02± 0.12,1.08± 0.10) were markedly lower than that of the LPS group(1.34± 0.12)(t=4.756,P< 0.01;t=2.235,2.308,P< 0.05).The TNF α levels were obviously lower in the low, middle and high dose inhibitory peptide groups[(93.66± 2.30), (75.78± 6.55), (71.50 ± 7.75) pg/L] than in the LPS group[(128.67± 39.67) pg/L] (t =2.216,P< 0.05;t=2.423,2.389,P< 0.01). CONCLUSION:LBP inhibitory peptide inhibits the NFκ B activity of U937 and the release of TNF α .LBP inhibitory peptide might have a potential value in preventing pulmonary injury induced by LPS.

关 键 词：脂多糖类 NF-κB 肿瘤坏死因子 

分 类 号：R563[医药卫生—呼吸系统]