贵州医科大学学报

2009, v.34;No.138(03) 262-266

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内毒素对离体鼻黏膜上皮细胞IFN-γ、IL-4表达的影响
The Effect of Endotoxin to Expression of IFN-γ and IL-4 in Human Nasal Epithelial Cell in vitro

杨秀海;李源;高晓勤;
YANG Xiuhai1,LI Yuan2,GAO Xiaoqin3 (1.Department of Otorhinolarynglogy and Cephalic-Cervical Surgery,Guizhou Provincial People's Hospital,Guiyang 550002,Guizhou,China;2.Department of Otorhinolarynglogy and Cephalic-Cervical Surgery,The Third Affiliated Hospital of Sun Yat-Sen University,Guangzhou 510630,Guangdong,China;3.Department of Histo-Embryology,Guiyang Medical college,Guiyang 550004,Guizhou,China;)

摘要(Abstract):

目的:探讨内毒素(LPS)对鼻黏膜上皮细胞(HNEC)IFN-γ、IL-4表达的影响。方法:设LPS组、LPS+地塞米松(DXM)组、LPS+吡咯啉烷二硫代氨基甲酸盐(PDTC)组及生理盐水空白对照组。采用酶联免疫吸附及原位杂交方法检测培养上清液中IFN-γ、IL-4及HNEC内NF-кB p50mRNA的表达,凝胶电泳迁移率检测NF-кB p50,观察PDTC及DXM对NF-кB p50活化抑制后IFN-γ和IL-4蛋白水平。结果:(1)0~100μg/L LPS随浓度的增加,IFN-γ分泌量增多;>100μg/L后,IFN-γ分泌量减少;100μg/LLPS刺激HNEC分泌IFN-γ的作用最强。0~20μg/L LPS随浓度的增加,IL-4分泌量增多;>20μg/L后,IFN-γ分泌量减少;20μg/L LPS刺激HNEC分泌IL-4的作用最强,40μg/L LPS刺激IFN-γ和IL-4的分泌有交叉,二者维持于一定的水平。(2)50μg/L LPS刺激HNEC后随时间延长IFN-γ分泌量逐渐增加,从4 h开始即有明显上升(P<0.05),8 h达到峰值;IL-4随时间延长分泌量逐渐增加,从2 h开始即有明显上升(P<0.05),6 h达到峰值。(3)PDTC和DXM均抑制NF-кB p50的表达和活性,减少HNEC分泌IFN-γ和IL-4,DXM4.0μg/L时,下降更明显,但高于此浓度时,不引起进一步的降低。结论:LPS对HNEC的刺激反应与浓度呈剂量依赖性,适当浓度的LPS刺激有利于HNEC分泌的Th1/Th2细胞因子维持平衡状态;过高或过低浓度的LPS刺激可诱导HNEC的Th1/Th2细胞因子失衡分泌,这可能是GNB在慢性鼻、鼻窦炎(CRS)的致病机理。
Objective: To study the influence of endoutoxin(lipopolysaccharide,LPS) to expression of IFN-γand IL-4 in human nasal epithelial cell(HNEC).Methods: Cultured HNEC were divided into group LPS,group DXM,and group PDTC,that were added with LPS,LPS plus dexamethasone,and LPS plus pyrroline dithiocarbamate respectively.A blank control group(group B) was set up by adding normal saline.Expression of IFN-γ and IL-4 in supernatants of culture media and NF-кB p50 mRNA expression in HNEC were detected with enzyme-linked immune adsorption(ELISA) and hybridization in situ;The activation of NF-кB p50 was detected by gel electrophoresis mobility analysis;The protein level changes of IFN-γ and IL-4 after the inhibition of PDTC and DXM to NF-кB p50 was observed.Result: LPS could directly act on the HNEC to promote the secretions of IFN-γ and IL-4 in terms of time.The LPS excitement could significantly promote the activation of NF-кB p50,which peaked in 3 hours and then went down.PDTC and DXM could significantly decrease analogically the secretions of IFN-γ and IL-4 and the activation of NF-кB p50.Conclusions: The response of HNEC to the stimulation of LPS shows dosage dependence,and stimulation of LPS with suitable concentration benefits HNEC secreting Th1/Th2 cell factors to maintain the balance;The stimulation of LPS with too high and too low concentrations would cause unbalanced secretion of Th1/Th2 cell factors,which might be the pathopoiesis mechanism of gram negative bacilli in chronic rhinitis and rhinosinusitis.

关键词(KeyWords): 内毒素类;上皮细胞;鼻黏膜;核因子кBp50;IFN-γ;白细胞介素4
endotoxins;epithelial cells;nasal mucosa;nucler factorкB p50;IFN-Υ;interleukin-4

Abstract:

Keywords:

基金项目(Foundation): 贵州省省长基金资助项目(20052A357)

作者(Author): 杨秀海;李源;高晓勤;
YANG Xiuhai1,LI Yuan2,GAO Xiaoqin3 (1.Department of Otorhinolarynglogy and Cephalic-Cervical Surgery,Guizhou Provincial People's Hospital,Guiyang 550002,Guizhou,China;2.Department of Otorhinolarynglogy and Cephalic-Cervical Surgery,The Third Affiliated Hospital of Sun Yat-Sen University,Guangzhou 510630,Guangdong,China;3.Department of Histo-Embryology,Guiyang Medical college,Guiyang 550004,Guizhou,China;)

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DOI: 10.19367/j.cnki.1000-2707.2009.03.008

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