三條脂多糖結合蛋白／CD14結合位點模擬肽體外抗內毒素活性的比較研究_《中國呼吸與危重監護雜志》

作者：

徐智 ¹ , 李琦 ¹ , 馮起甲 ¹ , 錢桂生 ¹ , 徐順貴 ¹ , 李昆霖 ² , 王興勝 ² ,  吳國明 ¹

1 第三軍醫大學新橋醫院全軍呼吸內科研究所全軍呼吸病研究重點實驗室(重慶400037);;
2 第三軍醫大學大坪醫院呼吸科;

關鍵詞：

脂多糖結合蛋白 CD14 模擬肽內毒素

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目的比較三條LBP／CD14結合位點模擬肽體外抑制內毒素性炎癥反應的生物學活性。方法采用酶聯免疫吸附法(ELISA)檢測模擬肽與CD14的親和力及與脂多糖結合蛋白(LBP)的競爭性抑制活性。將佛波脂(PMA)誘導成熟的U937細胞與內毒素共培養,并予以模擬肽干預,采用RT-PCR檢測模擬肽對U937細胞TNF. 基因表達的影響。將大鼠肺泡巨噬細胞(AMs)與熒光標記內毒素(FITC-LPS)共培養,并予以模擬肽干預,采用熒光顯微鏡觀測模擬肽對內毒素(LPS)與AMs結合的影響。結果 1號模擬肽(FHRWPTWPLPSP,10 μg／mL)與CD14的親和力顯著高于2號和3號模擬肽(20.3±4.1比11.8±2.4和13.7±3.3,P lt;0.01或P lt;0.05),1號模擬肽(10 μg／mL)對LBP的競爭性抑制活性也顯著高于2號和3號模擬肽[(57.2±11.2)％比(39.4±9.7)％和(37.9±8.3)％ ,P均 lt;0.01]。三條模擬肽(10 μg／mL)均可從基因水平顯著抑制LPS誘導U937細胞產生TNF-α(P lt;0.01或P lt;0.05),1號模擬肽的抑制作用顯著強于2號和3號模擬肽(0.239±.053比0.406±0.112和0.493±0.121,P lt;0.01)。1號模擬肽能顯著抑制LPS與AMs結合(2 157±514比2 763±453,P lt;0.01)。結論 1號模擬肽(FHRWPTWPLPSP)有較高的CD14親和力及LBP競爭性抑制活性,具有潛在的抗內毒素性炎癥反應的作用。

引用本文： 徐智,李琦,馮起甲,錢桂生,徐順貴,李昆霖,王興勝,吳國明. 三條脂多糖結合蛋白／CD14結合位點模擬肽體外抗內毒素活性的比較研究. 中國呼吸與危重監護雜志, 2008, 08(6): 435-439. doi: 復制

1.	Finberg RW ,Re F,Popova L,et al．Cell activation by Toll_like receptors:role of LBP and CD14．j Endotoxin Res,2004,10:413-418.
2.	Blais DR,Vascotto SG,Griffith M ,et al．LBP and CD14 secreted in tears by the lacrimal ands modulate the LPS response of corneal epithelial cells．Invest Ophthalmol Vis Sci,2005,46:4235-4244.
3.	Spek CA,Verbon A,Aberson H,et a1．Treatment with an anti-CD14 monoclonal antibody delays and inhibits lipopolysaccharide-induced gene expression in humans in vivo．J Clin Immunol,2003,23:132- 140.
4.	Kitchens RL,Thompson PA．Modulatory effects of sCD14 and LBP on LPS．host ce11 interactions．J Endotoxin Res,2005,11:225-229.
5.	馮起甲,徐智,吳國明,等．脂多糖結合蛋白與CD14結合位點模擬肽的初步篩選．第三軍醫大學學報,2007,29:1572-1575.
6.	敖啟林,黃磊,朱朋成,等．脂多糖活化的大鼠肺泡巨噬細胞TNF-α產生的新機制．中國病理生理雜志,2006,22:148-151.
7.	Kitchens RL,Thompson PA．Impact of sepsis induced changes in plasma on LPS interactions with monocytes and plasma lipoproteins: roles of soluble CD14,LBP,and acute phase lipoproteins．J Endotoxin Res,2003,9:113-118.
8.	Stein T,Morris JS,Davies CR,et al．Involution of the mouse mammary gland is associated with all immune cascade and an acute-phase response,involving LBP,CD14 and STAT3．Breast Cancer Res, 2004,6:R75-R91. 9 吳學玲,錢桂生,徐德斌,等．脂多糖結合蛋白抑制肽對內毒素誘導的人單核巨噬細胞株表達CD14的影響．中國呼吸與危重監護雜志,2005,4:230-232.
9.	Cunningham SC,Malone DL,Bochicchio GV,et al．Serum lipopolysaccharide-binding protein concentrations in trauma victims．Surg Infect,2006,7:251-261.
10.	Watkin RW ,Harper LV,Vernallis AB,et a1．Pro-inflammatory cytokines IL6,TNF-alpha,IL1 beta,procalcitonin,lipopolysaccharide binding protein and C-reactive protein in infective endocarditis．J Infeet,2007,55:220-225.
11.	Zweigner j,Schumann RR,Weber JR．The role of lipopotysaecha. ride- binding protein in modulating the innate immune response．Microbes Infect,2006,8:946-952.
12.	Iovine N,Eastvold J,Elsbach P,et al．The carboxyl-terminal domain of closely related endotoxin-binding proteins determ ines the target of protein-lipopolysaccharide complexes．J Biol Chem,2002,277:7970-7978.
13.	Samoylova TI,Morfison NE,Globa LP,et al．Peptide phage display: opportunities for development of personalized anti-cancer strate- gies．Anticancer Agents Med Chem,2006,6:9-17.
14.	Soykut EA,Dudak FC,Boyaei IH．Selection of staphylococcal entero. toxin B(SEB)-binding peptide using phage display technology．Bioehem Biophys Res Commun,2008,370:104-108．.

1. Finberg RW ,Re F,Popova L,et al．Cell activation by Toll_like receptors:role of LBP and CD14．j Endotoxin Res,2004,10:413-418.
2. Blais DR,Vascotto SG,Griffith M ,et al．LBP and CD14 secreted in tears by the lacrimal ands modulate the LPS response of corneal epithelial cells．Invest Ophthalmol Vis Sci,2005,46:4235-4244.
3. Spek CA,Verbon A,Aberson H,et a1．Treatment with an anti-CD14 monoclonal antibody delays and inhibits lipopolysaccharide-induced gene expression in humans in vivo．J Clin Immunol,2003,23:132- 140.
4. Kitchens RL,Thompson PA．Modulatory effects of sCD14 and LBP on LPS．host ce11 interactions．J Endotoxin Res,2005,11:225-229.
5. 馮起甲,徐智,吳國明,等．脂多糖結合蛋白與CD14結合位點模擬肽的初步篩選．第三軍醫大學學報,2007,29:1572-1575.
6. 敖啟林,黃磊,朱朋成,等．脂多糖活化的大鼠肺泡巨噬細胞TNF-α產生的新機制．中國病理生理雜志,2006,22:148-151.
7. Kitchens RL,Thompson PA．Impact of sepsis induced changes in plasma on LPS interactions with monocytes and plasma lipoproteins: roles of soluble CD14,LBP,and acute phase lipoproteins．J Endotoxin Res,2003,9:113-118.
8. Stein T,Morris JS,Davies CR,et al．Involution of the mouse mammary gland is associated with all immune cascade and an acute-phase response,involving LBP,CD14 and STAT3．Breast Cancer Res, 2004,6:R75-R91. 9 吳學玲,錢桂生,徐德斌,等．脂多糖結合蛋白抑制肽對內毒素誘導的人單核巨噬細胞株表達CD14的影響．中國呼吸與危重監護雜志,2005,4:230-232.
9. Cunningham SC,Malone DL,Bochicchio GV,et al．Serum lipopolysaccharide-binding protein concentrations in trauma victims．Surg Infect,2006,7:251-261.
10. Watkin RW ,Harper LV,Vernallis AB,et a1．Pro-inflammatory cytokines IL6,TNF-alpha,IL1 beta,procalcitonin,lipopolysaccharide binding protein and C-reactive protein in infective endocarditis．J Infeet,2007,55:220-225.
11. Zweigner j,Schumann RR,Weber JR．The role of lipopotysaecha. ride- binding protein in modulating the innate immune response．Microbes Infect,2006,8:946-952.
12. Iovine N,Eastvold J,Elsbach P,et al．The carboxyl-terminal domain of closely related endotoxin-binding proteins determ ines the target of protein-lipopolysaccharide complexes．J Biol Chem,2002,277:7970-7978.
13. Samoylova TI,Morfison NE,Globa LP,et al．Peptide phage display: opportunities for development of personalized anti-cancer strate- gies．Anticancer Agents Med Chem,2006,6:9-17.
14. Soykut EA,Dudak FC,Boyaei IH．Selection of staphylococcal entero. toxin B(SEB)-binding peptide using phage display technology．Bioehem Biophys Res Commun,2008,370:104-108．.

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三條脂多糖結合蛋白／CD14結合位點模擬肽體外抗內毒素活性的比較研究

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三條脂多糖結合蛋白／CD14結合位點模擬肽體外抗內毒素活性的比較研究

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