铜绿假单胞菌注射液对脓毒症大鼠的预防效应
|
摘要
革兰阴性菌感染引起的脓毒症在临床的死亡率很高,虽然人们对脓毒症的研究花费了大量的精力和金钱,但脓毒症的死亡率仍没有得到明显的改善。近年来人们发现TLR4 (toll-like-receptors 4)在革兰阴性菌感染中的免疫反应及脓毒症的全身炎症反应综合症(SIRS)中均起到了至关重要的作用。因此如何调节TLR4的活性来增强机体的免疫功能以达到控制G-细菌感染性疾病同时避免全身炎症反应综合症的发生成为现在研究的热点。铜绿假单胞菌注射液(PA-MSHA注射液)是一种基因工程技术制备的铜绿假单胞菌-甘露糖敏感血凝株菌毛灭活疫苗,因此可以作为G-细菌的病原体相关分子模式,来激活G-细菌的主要病原模式识别受体TLR4,从而有效活化人体免疫细胞,增强机体的免疫防御,已经广泛用于多种恶性肿瘤和感染性疾病的治疗。本课题首先观察预防应用不同剂量和给药时间的铜绿假单胞菌注射液干预后的S-D大鼠CLP术(盲肠结扎穿刺术)后的生存率变化,其次检测生存率有明显差异实验组大鼠不同时间点血液及组织标本中TLR4和TNF-αIL-1、IL-4、IL-6、IL-10炎症因子的水平。最后探讨大鼠脓毒症造模后的生存率和免疫水平、炎症因子及TLR4活化状态的关系。
目的：
1、评价预防性应用铜绿假单胞菌注射液对S-D大鼠CLP术后7天生存率的影响。
2、探讨药物干预后的大鼠CLP术后生存率与免疫状态,炎症因子及TLR4活化状态的关系。
方法：
雄性S-D大鼠,按照随机化原则分入实验A、B、C、D、E组及对照组,每组12只。各组的用药方法分别为：A组(PA-MSHA注射液0.25ml S. C(皮下注射)qod共8天)、B组(PA-MSHA注射液0.5ml S. C qod共8天)、C组(PA-MSHA注射液0.125ml S. C qod共16天)、D组(PA-MSHA注射液0.25ml S. C qod共16天)、E组(PA-MSHA注射液0.5ml S. C qod共16天)、对照组(空白对照,无任何干预)。各实验组用药结束后给予CLP造模,对照组无任何干预直接行CLP术。观察和记录术后7天各组大鼠的整体生活状态、临床评分、死亡数目和死亡时间,死亡鼠均做尸体解剖。
检测生存率观察中生存率最高实验组、生存率最低实验组用药后和CLP术后24小时血液和组织标本中TLR4, TNF-a、IL-1、IL-6、IL-4、IL-10等炎症相关因子表达情况。
结果：
1、生存率观察大鼠术后7天的生存率分别为：A组58.3%、B组91.7%、C组66.6%、D组70%、E组41.7%、对照组42.9%。生存率和生存曲线比较示：B组和对照组、B组和E组存在统计学差异(P<0.05)
2、实验组药物干预后TLR4和炎症因子水平与对照组相比较均有所增加,生存率最高组增加的非常明显,有统计学差异(P<0.05)。实验组CLP术后TLR4和炎症因子水平与对照组相比较均有所降低,生存率最低组降低的较明显,有统计学差异。
结论：
1、铜绿假单胞菌注射液一定条件下的预防性应用,是可以改善S-D大鼠CLP术后生存率。
2、S-D大鼠CLP术后生存率和术前免疫状态,炎症因子水平及TLR4活化状态是相关的。
Mortality rate of sepsis caused by gram-negative bacterium infection in clinical is very high, although people has spent a lot of energy and money on the research of sepsis, mortality rate of sepsis still remained no improved. In recent years, people found TLR4 play a very important role in the immune response of gram-negative bacterium infection and systemic inflammatory response syndrome (SIRS) of sepsis, so how to adjust the activity of TLR4, strengthen the immune function, to control the G-bacteria infectious diseases and to avoid SIRS became research hot spot today.
Pseudomonas aeruginosa injection (PA-MSHA injection) is a kind of Pseudomonas aeruginosa Mannose-sensitive hemagglutination inactivated vaccine made through genetic engineering technology. So it can activate G- bacterial pathogens related molecular mode receptor TLR4 as G- bacterial pathogens related molecular mode, thus effectively activate the body's immune cells, strengthen the body's immune defenses, it has been widely used in the treatment of various malignant tumor and infectious disease. Firstly, we observe the survival rate of rats with the preventive intervention of different doses and time pseudomonas aeruginosa injection after CLP operation. Secondly we detect the levels of TLR4 and inflammation factors such as TNF-a, IL-1, IL-4, IL-6, IL-10 in tissue and blood samples of the experimental groups significantly different in survival rate and control groups in different times. The paper also discussed the relationship among the survival rate of rats after CLP operation, immune level, inflammation factors and TLR4 activity state.
Obejective:
1、To evaluation the effect of Pseudomonas aeruginosa injection preventative intervention on the survival of the rats during the 7 days after CLP operation.
2、To discuss the relationship among the survival rate of rats after CLP operation, immune level, inflammation factors and TLR4 activity state.
Methods:
S-D male rats were divided into 5 experimental group A,B,C,D,E, and 1 control group at random,12 rats each group. The intervention of each experimental group are respectively:A(PA-MSHA injection 0.25 ml S.C qod for 8 days), B(PA-MSHA injection 0.5 ml S.C qod for 8 days), C(PA-MSHA injection 0.125 ml S.C qod for 16 days), D(PA-MSHA injection 0.25 ml S.C qod for 16 days), E(PA-MSHA injection 0.5 ml S.C qod for 16 days), control group(without any intervention).all rats were given CLP operation after intervention. The rats of control group were given CLP operation without any intervention. During the 7 days after CLP operation, all rats'physical activity, clinical score,the number of deaths and the time of deaths were observed and recorded, all rats underwent autopsy after death.
TLR4 and inflammation factors such as TNF-a, IL-1, IL-6, IL-4, IL-10 in the blood and tissue samples of survival supreme group, survival lowest group and control group was tested after intervention and 12 hours after CLP operation.
Results:
1、The survival rate of 6 groups 7 days after operation are respectively:group A 58.3%, group B 91.7%, group C 66.6%,group D 70%, group E 41.7%, control group 42.9%.There are significant statistically differences(P<0.05) between group B and control group, group B and group E, in the survival rate and the survival curves comparison.
2、The level of TLR4 and inflammation factors of experimental group after the intervention were both increased compared with control group, the increase of the survival supreme group was very significant, and has statistical difference compare with control group (P<0.05). The level of TLR4 and inflammation factors of experimental group after the CLP operation were both reduced compared with control group, the reduce of the survival lowest group was very significant, and has statistical difference compare with control group (P<0.05)
Conclusion:
1、The preventive application of PA-MSHA injection can improve the survival rate of S-D rats after CLP operation under certain conditions.
2、The survival rate of S-D rats after CLP operation is related with immune level, inflammation factors and the activity state of TLR4 before CLP operation.
目的：
1、评价预防性应用铜绿假单胞菌注射液对S-D大鼠CLP术后7天生存率的影响。
2、探讨药物干预后的大鼠CLP术后生存率与免疫状态,炎症因子及TLR4活化状态的关系。
方法：
雄性S-D大鼠,按照随机化原则分入实验A、B、C、D、E组及对照组,每组12只。各组的用药方法分别为：A组(PA-MSHA注射液0.25ml S. C(皮下注射)qod共8天)、B组(PA-MSHA注射液0.5ml S. C qod共8天)、C组(PA-MSHA注射液0.125ml S. C qod共16天)、D组(PA-MSHA注射液0.25ml S. C qod共16天)、E组(PA-MSHA注射液0.5ml S. C qod共16天)、对照组(空白对照,无任何干预)。各实验组用药结束后给予CLP造模,对照组无任何干预直接行CLP术。观察和记录术后7天各组大鼠的整体生活状态、临床评分、死亡数目和死亡时间,死亡鼠均做尸体解剖。
检测生存率观察中生存率最高实验组、生存率最低实验组用药后和CLP术后24小时血液和组织标本中TLR4, TNF-a、IL-1、IL-6、IL-4、IL-10等炎症相关因子表达情况。
结果：
1、生存率观察大鼠术后7天的生存率分别为：A组58.3%、B组91.7%、C组66.6%、D组70%、E组41.7%、对照组42.9%。生存率和生存曲线比较示：B组和对照组、B组和E组存在统计学差异(P<0.05)
2、实验组药物干预后TLR4和炎症因子水平与对照组相比较均有所增加,生存率最高组增加的非常明显,有统计学差异(P<0.05)。实验组CLP术后TLR4和炎症因子水平与对照组相比较均有所降低,生存率最低组降低的较明显,有统计学差异。
结论：
1、铜绿假单胞菌注射液一定条件下的预防性应用,是可以改善S-D大鼠CLP术后生存率。
2、S-D大鼠CLP术后生存率和术前免疫状态,炎症因子水平及TLR4活化状态是相关的。
Mortality rate of sepsis caused by gram-negative bacterium infection in clinical is very high, although people has spent a lot of energy and money on the research of sepsis, mortality rate of sepsis still remained no improved. In recent years, people found TLR4 play a very important role in the immune response of gram-negative bacterium infection and systemic inflammatory response syndrome (SIRS) of sepsis, so how to adjust the activity of TLR4, strengthen the immune function, to control the G-bacteria infectious diseases and to avoid SIRS became research hot spot today.
Pseudomonas aeruginosa injection (PA-MSHA injection) is a kind of Pseudomonas aeruginosa Mannose-sensitive hemagglutination inactivated vaccine made through genetic engineering technology. So it can activate G- bacterial pathogens related molecular mode receptor TLR4 as G- bacterial pathogens related molecular mode, thus effectively activate the body's immune cells, strengthen the body's immune defenses, it has been widely used in the treatment of various malignant tumor and infectious disease. Firstly, we observe the survival rate of rats with the preventive intervention of different doses and time pseudomonas aeruginosa injection after CLP operation. Secondly we detect the levels of TLR4 and inflammation factors such as TNF-a, IL-1, IL-4, IL-6, IL-10 in tissue and blood samples of the experimental groups significantly different in survival rate and control groups in different times. The paper also discussed the relationship among the survival rate of rats after CLP operation, immune level, inflammation factors and TLR4 activity state.
Obejective:
1、To evaluation the effect of Pseudomonas aeruginosa injection preventative intervention on the survival of the rats during the 7 days after CLP operation.
2、To discuss the relationship among the survival rate of rats after CLP operation, immune level, inflammation factors and TLR4 activity state.
Methods:
S-D male rats were divided into 5 experimental group A,B,C,D,E, and 1 control group at random,12 rats each group. The intervention of each experimental group are respectively:A(PA-MSHA injection 0.25 ml S.C qod for 8 days), B(PA-MSHA injection 0.5 ml S.C qod for 8 days), C(PA-MSHA injection 0.125 ml S.C qod for 16 days), D(PA-MSHA injection 0.25 ml S.C qod for 16 days), E(PA-MSHA injection 0.5 ml S.C qod for 16 days), control group(without any intervention).all rats were given CLP operation after intervention. The rats of control group were given CLP operation without any intervention. During the 7 days after CLP operation, all rats'physical activity, clinical score,the number of deaths and the time of deaths were observed and recorded, all rats underwent autopsy after death.
TLR4 and inflammation factors such as TNF-a, IL-1, IL-6, IL-4, IL-10 in the blood and tissue samples of survival supreme group, survival lowest group and control group was tested after intervention and 12 hours after CLP operation.
Results:
1、The survival rate of 6 groups 7 days after operation are respectively:group A 58.3%, group B 91.7%, group C 66.6%,group D 70%, group E 41.7%, control group 42.9%.There are significant statistically differences(P<0.05) between group B and control group, group B and group E, in the survival rate and the survival curves comparison.
2、The level of TLR4 and inflammation factors of experimental group after the intervention were both increased compared with control group, the increase of the survival supreme group was very significant, and has statistical difference compare with control group (P<0.05). The level of TLR4 and inflammation factors of experimental group after the CLP operation were both reduced compared with control group, the reduce of the survival lowest group was very significant, and has statistical difference compare with control group (P<0.05)
Conclusion:
1、The preventive application of PA-MSHA injection can improve the survival rate of S-D rats after CLP operation under certain conditions.
2、The survival rate of S-D rats after CLP operation is related with immune level, inflammation factors and the activity state of TLR4 before CLP operation.
引文
[1]Smith CR, Stranhe RC, Ziegler EJ, et al. HA-IA, a human monoclonal antibody for the treatment of gram- negative sepsis. Infect Dis Clin North Am,1992,6: 253.
[2]谷元廷,吴河水,徐建波,等.全肝缺血再灌注小鼠肺泡巨噬细胞Toll样受体2/4的表达[J].郑州大学学报：医学版,2006,41(6)：1047-1049.
[3]TSUJMOTO H, ONO S, EFRON P A, et al Role of toll-like receptors in the development of sepsis[J]. Shock,2008,29(3):315-321.
[4]Forster Waldl E. Sadeghi K, Tamandl D, et al Monocyte toll-like receptor 4 expression and LPS-induced cytokine production increase.
[5]车兆义,邹悦,宋清斌 大鼠实验中几种常用的采血方法探讨[J]局解手术学杂志.2008,17(2)：84-85.
[6]盛志勇,姚咏明.脓毒症研究的现状与展望.解放军医学杂志,1999,24：79-82.
Sheng ZY, Yao YM. The presenl and prospects of sepsis research. J PLA,1999, 24:79-82.
[7]姚咏明,盛志勇.我国创伤脓毒症基础研究新进展.中华创伤杂志,200319：9-12.
Yao YM, Sheng ZY. New advances in basic scientific research on sepsis in China. Chin J Trauma,2003,19:9-12.
[8]Martin Gs, Mannino DM, Eaton S, et al. The epidemiology of sepsis in the United States from 1979 through 2000 [J]. N Engl J Med,2003, 348(16):1546-1554.
[9]Angus DC, Linde-Zwirble WT, Lidicker J, et al. Epidemiology of severe sepsis in the Unite State:analysis of incidence, outcome, and associated costs of care[J]. Crit Care Med,2001,29(7):1303-1310.
[10]盛志勇,姚咏明。脓毒症与多器官功能障碍综合症[J].中华急诊医学杂志,2003,12(10)：643-654.
[11]FRANTZS, ERTLG, BAUERSACHS J. Mechanisms of disease:Toll-like receptors in cardiovascular disease [J]. Nat Clin Pract Cardi ovascMed,2007, 4(8):444-454.
[12]Levy MM, Fink MP, Marshall JC, et al.2001 SCCM/ESICM/ACCP/ATS/SIS international Sepsis Definitions Conference [J]. Crit Care Med,2003,31:1250-1256
[13]Peres-Bota D, Melot C, Lopes F, et al. Infection probability score (IPS):a simple method to help assess the probability of infection in critically patients [J]. Crit Care Med,2003,31:2579-2584
[14]Cao Z, Shi L, Li Y, et al. Pseudomonas aeruginosa:Mannose Sensitive Hemagglutinin Inhibits the Growth of Human Hepatocarcinoma Cells via Mannose Mediated Apoptosis [J]. Digestive Diseases and Sciences,2009 54(10):211-227.
[15]凌伟,刘骅,曹晖,等.铜绿假单胞菌注射液对胃癌细胞体外杀伤作用的观察[J].中华肿瘤防治杂志,2008,15(18)：1381-1385.
[16]Liu Zhebin, Hou Yifeng, Dong Min, et al. PA-MSHA inhibit s proliferation and induces apoptosis through the up-regulation and activation of caspases in the human breast cancer cell lines [J]. J Cell Biochem,2009,108 (2):195-206.
[17]刘奎杰,赵华,雷三林.铜绿假单胞菌注射液对胃肠道肿瘤患者围手术期NK细胞调节[J].中南药学,2009,7(5)：392-394.
[18]孙文平,牟希亚,高小平,等.铜绿假单胞菌菌苗对尿路感染的免疫功能调节[J].微生物学杂志,2007,22(4)：20-22.
[19]单红卫,林兆奋,赵良,等.PA-MSHA疫苗对严重创伤患者并发肺部感染的预防作用[J].中国急救医学,2007,27(4)：375-377.
[20]Mossman K L, MianMF, Lauzon N M, et al. Cutting edge:FimH adhesin of type 1 flmbriae is a novel TLR4 ligand[J]. J Immunol,2008.81(10):6702-6706.
[21]孙文平,付红文,刘妮,等.PA-MSHA菌毛株疫苗对三种癌症患者免疫疗效的临床观察[J].中华微生物学和免疫学杂志,2000,20(4)373-376.
[22]Ecker M, Muller G. Cytokine release after administration of endotoxin containing vaccines [J]. A L T EX,1998,15:68-71
[23]李听,刘佳佳.IL-6对中性粒细胞在炎症中作用的影响[J].国外医学免疫学分册,2005,28(9)：277-280.
[24]蔡美英主编.医学免疫学[M].北京：科学出版社,51.
[25]沈蕾.脓毒症大鼠组织c- FLIP的变化及乌司他丁对其的影响(D).上海：复旦大学,2008：
[26]姚咏明.重视对脓毒症免疫状态的监测与评估[J].中华急诊医学杂志,2007,16：795-796.
[1]Smith CR, Stranhe RC, Ziegler EJ, et al. HA-IA, a human monoclonal antibody for the treatment of gram-negative sepsis. Infect Dis Clin North Am,1992,6: 253.
[2]谷元廷,吴河水,徐建波,等.全肝缺血再灌注小鼠肺泡巨噬细胞Toll样受体2/4的表达[J].郑州大学学报：医学版,2006,41(6)：1047-1049.
[3]TSUJMOTO H, ONOS, EFRON P A, et al Role of toll - like receptors in the development of sepsis [J]. Shock,2008,29(3):315-321.
[4]Forster Waldl E. Sadeghi K, Tamandl D, et al Monocyte toll - like receptor 4 expression and LPS-induced cytokine production increase during gestational aging[J].Pediatr Res,2005,58(1):121-124.
[5]BACHAR O, ADNER M, UDDMAN R, et al. Toll-like receptor stimulation induces airway hyper-responsiveness to bradykinin, an effect mediated by JNK and NF- kappa B signaling pathways [J]. Eur J Immunol,2004,34 (4):1196-1207.
[6]VERSTAK B, HERTZOG P, MANSELL A. Toll-like receptor signalling and the clinical benefits that lie within [J].Inflamm Res,2007,56(1):1-10.
[7]Perez-Vilar J, Mabolo R, McVaugh CT, et al Mucin granule intraluminal organization in living mucous/goblet cells Roles of protein post-translational modifications and secretion [J].JB iol Chem,2006,281 (8):4844-4855.
[8]Means TK, Golenbock DT, Fent onMJ. The biol ogy of Toll-like receptors [J]. Cytokine Grow th Factor Rev,2000,11(3):219-232.
[9]Kawai T, Akira S. TLR signaling [J]. Cell Death Difer,2006,13 (5):816-825.
[10]Pearl-Yafe M, Fabian I, Halperin D, et al Interferon - gamma and bacterial lipopolysaccharide act synergistically on human neutropenhancing interleukin-8, interleukin-1 beta, tumor necrosis factoalpha, and interleukin-12 p70 secretion and phagocytosis via up relation of toll-like receptor 4 [J]. Shock,2007, 27(3):226-231.
[11]Yoshioka M, Fukuishi N, Kubo Y, et al Human cathelicidin CAP18/LL-37 changes mast cell function toward innate immunity [J]. B iol Pharm Bull, 2008,31(2):212-216。
[12]Qin J, Qian Y, Yao J, et al SIGIRR inhibits interleukin-1 receptor and toll-like receptor4-mediated signaling through different mechanisms [J]. J Biol Chem,2005, 280(26):25233-25241.
[13]Kobayashi K, Hernandez LD, Gala n JE, et al. I RAK2 M is a negative regulator of Toll-like receptor signaling [J]. Cell,2002,110(2):192-202.
[14]Zhang G, Ghosh S. Negative regulation of toll-like receptor mediated signaling by Tollip [J]. J Biol Chem,2002,277 (9):7059-7065.
[15]Wang Y, Tang Y, Teng L, et al. Association of beta-arrestin and TRAF6 negatively regulates Toll-like receptor interleukin 1 receptor signaling [J]. Nat Immunol,2006,7(2):139-147.
[16]Dagvadorj J, Naiki Y, Tumurkhuu G, et al. Interleukin-10 inhibits tumor necrosis factor- production in lipopolysaccharide-stimulated RAW 264.7 cells through reduced My D88 expression [J]. Innate Immunity,2008,14:109-115.
[17]Shimazu R, Akashi S, Ogata H, et al MD-2, a molecule that confers lipopolysaccharide responsiveness on toll-like receptor 4 J Exp Med,1999, 189:1777-1782.
[18]Qureshi ST, Lariviere L, Leveque G, et al. Endotoxin-toler-ant mice have mutations in toll-like receptor 4 (Tlr4). J Exp Med,1999,189(4):615-25.
[19]蒋建新,朱佩芳,王正国.内毒素基因与Toll样受体4基因致病机制的研究进展.中华创伤杂志,2002,18(12)：765-7.
[20]Arbour NC, Lorenz E, Schutte BC, et al. TLR4 mutations are associated with endotoxin hyporesponsiveness in humans Nat Genet,2000,25:187-191.
[21]Agnese DM, Calvano JE, Hahm SJ, et al Human toll-like receptor 4 mutations but not CD14 polymorphisms are associated with an increased risk of gram-negative infections [J]. J InfectD is,2002,186(2):1522-1525.
[22]Hawn TR, Verbon A, JanerM, et al Toll-like receptor-4 polymorphisms are associated with resistance to Legionnaires' disease [J]. Proc Natl Acad Sci USA, 2005,102 (6):2487-2489.
[23]SHA T, SUNAMOTOM, KITAZAKI T, et al. Therapeutic effects of TAK-242, a novel selective Toll-like receptor 4 signal transducti on inhibitor, in mouse endotoxin shock model [J]. Eur JPharmacol,2007,571 (2-3):231-239.
[24]NAK AMURA M, SH I MIZU Y, SATO Y, et al. Toll-like recept or 4 signal transducti on inhibitor, M62812, suppresses endothelial cell and leukocyte activation and prevents lethal septic shock inmice[J]. Eur J Phar macol,2007,569(3):237-243.
[2]谷元廷,吴河水,徐建波,等.全肝缺血再灌注小鼠肺泡巨噬细胞Toll样受体2/4的表达[J].郑州大学学报：医学版,2006,41(6)：1047-1049.
[3]TSUJMOTO H, ONO S, EFRON P A, et al Role of toll-like receptors in the development of sepsis[J]. Shock,2008,29(3):315-321.
[4]Forster Waldl E. Sadeghi K, Tamandl D, et al Monocyte toll-like receptor 4 expression and LPS-induced cytokine production increase.
[5]车兆义,邹悦,宋清斌 大鼠实验中几种常用的采血方法探讨[J]局解手术学杂志.2008,17(2)：84-85.
[6]盛志勇,姚咏明.脓毒症研究的现状与展望.解放军医学杂志,1999,24：79-82.
Sheng ZY, Yao YM. The presenl and prospects of sepsis research. J PLA,1999, 24:79-82.
[7]姚咏明,盛志勇.我国创伤脓毒症基础研究新进展.中华创伤杂志,200319：9-12.
Yao YM, Sheng ZY. New advances in basic scientific research on sepsis in China. Chin J Trauma,2003,19:9-12.
[8]Martin Gs, Mannino DM, Eaton S, et al. The epidemiology of sepsis in the United States from 1979 through 2000 [J]. N Engl J Med,2003, 348(16):1546-1554.
[9]Angus DC, Linde-Zwirble WT, Lidicker J, et al. Epidemiology of severe sepsis in the Unite State:analysis of incidence, outcome, and associated costs of care[J]. Crit Care Med,2001,29(7):1303-1310.
[10]盛志勇,姚咏明。脓毒症与多器官功能障碍综合症[J].中华急诊医学杂志,2003,12(10)：643-654.
[11]FRANTZS, ERTLG, BAUERSACHS J. Mechanisms of disease:Toll-like receptors in cardiovascular disease [J]. Nat Clin Pract Cardi ovascMed,2007, 4(8):444-454.
[12]Levy MM, Fink MP, Marshall JC, et al.2001 SCCM/ESICM/ACCP/ATS/SIS international Sepsis Definitions Conference [J]. Crit Care Med,2003,31:1250-1256
[13]Peres-Bota D, Melot C, Lopes F, et al. Infection probability score (IPS):a simple method to help assess the probability of infection in critically patients [J]. Crit Care Med,2003,31:2579-2584
[14]Cao Z, Shi L, Li Y, et al. Pseudomonas aeruginosa:Mannose Sensitive Hemagglutinin Inhibits the Growth of Human Hepatocarcinoma Cells via Mannose Mediated Apoptosis [J]. Digestive Diseases and Sciences,2009 54(10):211-227.
[15]凌伟,刘骅,曹晖,等.铜绿假单胞菌注射液对胃癌细胞体外杀伤作用的观察[J].中华肿瘤防治杂志,2008,15(18)：1381-1385.
[16]Liu Zhebin, Hou Yifeng, Dong Min, et al. PA-MSHA inhibit s proliferation and induces apoptosis through the up-regulation and activation of caspases in the human breast cancer cell lines [J]. J Cell Biochem,2009,108 (2):195-206.
[17]刘奎杰,赵华,雷三林.铜绿假单胞菌注射液对胃肠道肿瘤患者围手术期NK细胞调节[J].中南药学,2009,7(5)：392-394.
[18]孙文平,牟希亚,高小平,等.铜绿假单胞菌菌苗对尿路感染的免疫功能调节[J].微生物学杂志,2007,22(4)：20-22.
[19]单红卫,林兆奋,赵良,等.PA-MSHA疫苗对严重创伤患者并发肺部感染的预防作用[J].中国急救医学,2007,27(4)：375-377.
[20]Mossman K L, MianMF, Lauzon N M, et al. Cutting edge:FimH adhesin of type 1 flmbriae is a novel TLR4 ligand[J]. J Immunol,2008.81(10):6702-6706.
[21]孙文平,付红文,刘妮,等.PA-MSHA菌毛株疫苗对三种癌症患者免疫疗效的临床观察[J].中华微生物学和免疫学杂志,2000,20(4)373-376.
[22]Ecker M, Muller G. Cytokine release after administration of endotoxin containing vaccines [J]. A L T EX,1998,15:68-71
[23]李听,刘佳佳.IL-6对中性粒细胞在炎症中作用的影响[J].国外医学免疫学分册,2005,28(9)：277-280.
[24]蔡美英主编.医学免疫学[M].北京：科学出版社,51.
[25]沈蕾.脓毒症大鼠组织c- FLIP的变化及乌司他丁对其的影响(D).上海：复旦大学,2008：
[26]姚咏明.重视对脓毒症免疫状态的监测与评估[J].中华急诊医学杂志,2007,16：795-796.
[1]Smith CR, Stranhe RC, Ziegler EJ, et al. HA-IA, a human monoclonal antibody for the treatment of gram-negative sepsis. Infect Dis Clin North Am,1992,6: 253.
[2]谷元廷,吴河水,徐建波,等.全肝缺血再灌注小鼠肺泡巨噬细胞Toll样受体2/4的表达[J].郑州大学学报：医学版,2006,41(6)：1047-1049.
[3]TSUJMOTO H, ONOS, EFRON P A, et al Role of toll - like receptors in the development of sepsis [J]. Shock,2008,29(3):315-321.
[4]Forster Waldl E. Sadeghi K, Tamandl D, et al Monocyte toll - like receptor 4 expression and LPS-induced cytokine production increase during gestational aging[J].Pediatr Res,2005,58(1):121-124.
[5]BACHAR O, ADNER M, UDDMAN R, et al. Toll-like receptor stimulation induces airway hyper-responsiveness to bradykinin, an effect mediated by JNK and NF- kappa B signaling pathways [J]. Eur J Immunol,2004,34 (4):1196-1207.
[6]VERSTAK B, HERTZOG P, MANSELL A. Toll-like receptor signalling and the clinical benefits that lie within [J].Inflamm Res,2007,56(1):1-10.
[7]Perez-Vilar J, Mabolo R, McVaugh CT, et al Mucin granule intraluminal organization in living mucous/goblet cells Roles of protein post-translational modifications and secretion [J].JB iol Chem,2006,281 (8):4844-4855.
[8]Means TK, Golenbock DT, Fent onMJ. The biol ogy of Toll-like receptors [J]. Cytokine Grow th Factor Rev,2000,11(3):219-232.
[9]Kawai T, Akira S. TLR signaling [J]. Cell Death Difer,2006,13 (5):816-825.
[10]Pearl-Yafe M, Fabian I, Halperin D, et al Interferon - gamma and bacterial lipopolysaccharide act synergistically on human neutropenhancing interleukin-8, interleukin-1 beta, tumor necrosis factoalpha, and interleukin-12 p70 secretion and phagocytosis via up relation of toll-like receptor 4 [J]. Shock,2007, 27(3):226-231.
[11]Yoshioka M, Fukuishi N, Kubo Y, et al Human cathelicidin CAP18/LL-37 changes mast cell function toward innate immunity [J]. B iol Pharm Bull, 2008,31(2):212-216。
[12]Qin J, Qian Y, Yao J, et al SIGIRR inhibits interleukin-1 receptor and toll-like receptor4-mediated signaling through different mechanisms [J]. J Biol Chem,2005, 280(26):25233-25241.
[13]Kobayashi K, Hernandez LD, Gala n JE, et al. I RAK2 M is a negative regulator of Toll-like receptor signaling [J]. Cell,2002,110(2):192-202.
[14]Zhang G, Ghosh S. Negative regulation of toll-like receptor mediated signaling by Tollip [J]. J Biol Chem,2002,277 (9):7059-7065.
[15]Wang Y, Tang Y, Teng L, et al. Association of beta-arrestin and TRAF6 negatively regulates Toll-like receptor interleukin 1 receptor signaling [J]. Nat Immunol,2006,7(2):139-147.
[16]Dagvadorj J, Naiki Y, Tumurkhuu G, et al. Interleukin-10 inhibits tumor necrosis factor- production in lipopolysaccharide-stimulated RAW 264.7 cells through reduced My D88 expression [J]. Innate Immunity,2008,14:109-115.
[17]Shimazu R, Akashi S, Ogata H, et al MD-2, a molecule that confers lipopolysaccharide responsiveness on toll-like receptor 4 J Exp Med,1999, 189:1777-1782.
[18]Qureshi ST, Lariviere L, Leveque G, et al. Endotoxin-toler-ant mice have mutations in toll-like receptor 4 (Tlr4). J Exp Med,1999,189(4):615-25.
[19]蒋建新,朱佩芳,王正国.内毒素基因与Toll样受体4基因致病机制的研究进展.中华创伤杂志,2002,18(12)：765-7.
[20]Arbour NC, Lorenz E, Schutte BC, et al. TLR4 mutations are associated with endotoxin hyporesponsiveness in humans Nat Genet,2000,25:187-191.
[21]Agnese DM, Calvano JE, Hahm SJ, et al Human toll-like receptor 4 mutations but not CD14 polymorphisms are associated with an increased risk of gram-negative infections [J]. J InfectD is,2002,186(2):1522-1525.
[22]Hawn TR, Verbon A, JanerM, et al Toll-like receptor-4 polymorphisms are associated with resistance to Legionnaires' disease [J]. Proc Natl Acad Sci USA, 2005,102 (6):2487-2489.
[23]SHA T, SUNAMOTOM, KITAZAKI T, et al. Therapeutic effects of TAK-242, a novel selective Toll-like receptor 4 signal transducti on inhibitor, in mouse endotoxin shock model [J]. Eur JPharmacol,2007,571 (2-3):231-239.
[24]NAK AMURA M, SH I MIZU Y, SATO Y, et al. Toll-like recept or 4 signal transducti on inhibitor, M62812, suppresses endothelial cell and leukocyte activation and prevents lethal septic shock inmice[J]. Eur J Phar macol,2007,569(3):237-243.