目的 比較三條LBP／CD14結(jié)合位點模擬肽體外抑制內(nèi)毒素性炎癥反應(yīng)的生物學(xué)活性。方法 采用酶聯(lián)免疫吸附法(ELISA)檢測模擬肽與CD14的親和力及與脂多糖結(jié)合蛋白(LBP)的競爭性抑制活性。將佛波脂(PMA)誘導(dǎo)成熟的U937細(xì)胞與內(nèi)毒素共培養(yǎng),并予以模擬肽干預(yù),采用RT-PCR檢測模擬肽對U937細(xì)胞TNF. 基因表達(dá)的影響。將大鼠肺泡巨噬細(xì)胞(AMs)與熒光標(biāo)記內(nèi)毒素(FITC-LPS)共培養(yǎng),并予以模擬肽干預(yù),采用熒光顯微鏡觀測模擬肽對內(nèi)毒素(LPS)與AMs結(jié)合的影響。結(jié)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(dǎo)U937細(xì)胞產(chǎn)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(qiáng)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(jié)合(2 157±514比2 763±453,P lt;0.01)。結(jié)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(nèi)毒素性炎癥反應(yīng)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(jié)合蛋白／CD14結(jié)合位點模擬肽體外抗內(nèi)毒素活性的比較研究. 中國呼吸與危重監(jiān)護(hù)雜志, 2008, 08(6): 435-439. doi: 復(fù)制
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| 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. |
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| 5. | 馮起甲,徐智,吳國明,等．脂多糖結(jié)合蛋白與CD14結(jié)合位點模擬肽的初步篩選．第三軍醫(yī)大學(xué)學(xué)報,2007,29:1572-1575. |
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| 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(jié)合蛋白與CD14結(jié)合位點模擬肽的初步篩選．第三軍醫(yī)大學(xué)學(xué)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(xì)胞TNF-α產(chǎn)生的新機(jī)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(xué)玲,錢桂生,徐德斌,等．脂多糖結(jié)合蛋白抑制肽對內(nèi)毒素誘導(dǎo)的人單核巨噬細(xì)胞株表達(dá)CD14的影響．中國呼吸與危重監(jiān)護(hù)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．.