金银花提取物对小鼠T细胞行为的影响及脓毒症的免疫调节作用
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摘要
目的
研究金银花(Flos lonicerae,FL)提取物对小鼠T细胞行为和脓毒症的影响,探讨其对脓毒症小鼠的保护作用机制,为其在临床的应用提供参考数据和理论依据。
方法
1.体外结合流式细胞术利用荧光标记的单克隆抗体染色,检测FL对多克隆刺激剂刀豆蛋白A(Concanavalin A,Con A)刺激的小鼠淋巴细胞活化的影响;利用羧基荧光素二乙酸琥珀酰亚胺酯(CFDA-SE)染色分析FL对淋巴细胞增殖情况的影响;碘化丙锭(PI)染色分析FL对淋巴细胞周期分布的影响。
2.腹腔注射大肠杆菌(escherichia coli,E.coli)建立脓毒症模型,观察FL对小鼠生存状况的影响,分析各处理组小鼠胸腺、脾脏细胞总数,计算小鼠胸腺和脾脏指数,AnnexinV-FITC/PI双染检测FL对小鼠胸腺细胞凋亡的影响,使用CFDA-SE标记的大肠杆菌(E.Coli)结合流式细胞术检测FL对小鼠腹腔Mφ吞噬功能的影响。
3.利用腹腔注射LPS/D-半乳糖建立内毒素诱导脓毒症模型,采用MTT比色法检测FL对脓毒症小鼠淋巴细胞增殖的影响,通过H_2DCFDA染色检测外周血反应性氧族(reactive oxygen species,ROS)变化,利用H_2O_2测定试剂盒检测小鼠血清中H_2O_2的水平,通过血液生化和病理组织学检查,观察FL对小鼠肝功能的保护作用,结合体外试验,利用Griess反应试剂盒检测FL对脂多糖(LPS)激活的Mφ释放NO的影响。
结果
1.细胞活化分析结果表明,金银花提取物对小鼠淋巴细胞活化和增殖均有明显的抑制作用(P＜0.05),并且能明显阻止淋巴细胞进入S期和G2/M期。
2.大肠杆菌诱导的脓毒症小鼠胸腺和脾脏出现显著的萎缩现象;与此同时,药物治疗组小鼠胸腺和脾脏指数明显增加,结合体外实验,FL对DEX诱导的小鼠胸腺细胞凋亡都有显著的拮抗作用(P＜0.01);小鼠存活率亦显著提高(P＜0.01);腹腔巨噬细胞的吞噬功能明显增强。
3.利用内毒素诱导D-半乳糖致敏脓毒症模型,结果分析表明,与正常组相比,淋巴细胞对Con A诱导应答能力明显下降,外周血中性粒细胞ROS显著升高,血清中ALT、AST和H_2O_2水平亦增加,肝组织炎性细胞水平显著升高,肝脏出现病理损伤,而药物治疗组对增殖反应明显强于单纯模型组,外周血中性粒细胞ROS、血清中ALT、AST和H_2O_2水平均显著降低,肝组织炎性细胞水平减轻,Griess反应检测结果显示,FL明显抑制LPS激活巨噬细胞释放NO的量(P＜0.05)。
结论
1.FL能明显抑制小鼠淋巴细胞的体外活化和增殖,并能抑制淋巴细胞进入细胞分裂周期,提示FL对特异性细胞免疫有重要抑制作用,这可能是FL作为临床常用药物治疗免疫相关疾病的理论基础。
2.抑制胸腺和脾脏细胞凋亡和增强腹腔巨噬细胞吞噬功能可能FL对大肠杆菌所致脓毒症小鼠的保护机制。
3.清除氧自由基和提高脓毒症小鼠淋巴细胞免疫应答能力可能是FL对内毒素诱导的脓毒症小鼠的免疫保护机制。
Aim:
To investigate the effect of Flos lonicerae on mouse T lymphocytes behavior and sepsis,and elucidate the protective mechanism,in order to supply theory to clinical therapy.
Methods:
1.Lymphocytes isolated from lymphoid nodes of BalB/c mouse were stained with fluorescence conjugated monoclonal antibodies and flow cytometry analysis was employed to detect the effects of FL on the activation of mouse lymphocytes treated with Con A.The effects of FL on the proliferation rate of mouse lymphocytes were also analyzed by CFDA-SE staining.The distribution of the cell-cycle was analyzed by PI staining together with flow cytometry analysis.
2.For experiment sepsis,mouse was intraperitoneal injection(i.p.) of a bacterial suspension containing 10~8 live escherichia coli(E.coli) cells.We monitored the effect of FL on the mouse mortality rate in the experimental sepsis.The spleens and thymi were removed and cell yield was counted,and we compared spleen index and thymi index of different groups.Apoptosis of thymocytes DEX were detected by AnnexinV-FITC/PI staining together with flow cytometry.E.coli stained by CFDA-SE and flow cytometry was utilized to observe the effect of FL on macrophage phagocytosis.
3.For LPS-induced experimental sepsis,mice were given intraperitoneal injections of a mixture of LPS and D-galactosamine.The proliferation related index of mouse lymphocytes was analyzed with flow cytometry and MTT.Changes of reactive oxygen species(ROS) in blood were analyzed by H_2DCFDA staining together with flow cytometry.Serum ALT,AST levels and the pathological changes were obtained to study the protective of FL on liver function of septic mouse.Concentration of H_2O_2 in serum was measured by H_2O_2 kit.The effect of FL on NO production of the peritoneal macrophage in vitro was measured by Griess Reagent assay kit.
Results:
1.The results indicated that FL ihibited the activation of mouse lymphocytes(P＜0.05).CFDA-SE analysis also showed the proliferation of mouse lymphocytes were obviously inhibited by different concentrations of FL.Flow cytometry analysis of the cell-cycle distribution revealed that the cell number in G0/G1 was increased,while the cell number in S and G2/M was decreased as the concentration of FL increased.
2.The results demonstrated that septic mouse induced by the intraperitoneal injection of E.coli led to splenic atrophy.However,compared with septic mouse,FL could significantly increase thymus index and spleen index.FL could inhibit the apoptosis of thymus cells induced by Dex(P＜0.01).The survival rate of FL group was much higher than the sepsis group.FL promoted the phagocytosis.
3.Establishing sepsis model by intraperitoneal injections of LPS was to analysis the protective mechanism of FL on septic mouse.Compared with the control group,the lymphocytes proliferation reponses of septic mouse were reduced upon stimulation with Con A,and the levels of ALT,AST and H_2O_2 in serum and ROS in blood were increased significantly compared with the normal group,Meanwhile,the levels of inflammatory mediators were increased and morphological structures were damaged..But in the interference group FL could enhance proliferation of lymphocytes,and inhibit the levels of ALT,AST and H_2O_2 in serum and ROS in blood.Moreover,the levels of inflammatory mediators were significantly lower than septic group.FL could inhibit NO production of peritoneal macrophage induced by LPS with Griess Reagent assay kit.
Conclusion:
1.FL could significantly inhibit the activation and proliferation of mouse lymphocytes in vitro.Flow cytometry analysis of the cell-cycle distribution revealed that FL can inhibit the progression of mouse lymphocytes growing into S and G2/M phase. That indicated FL inhibited specific cellular immune response.These results supply theory to clinical therapy.
2.FL could inhibit atrophy in spleen and thymus induced by sepsis.FL could inhibit the apoptosis of thymus cells in vitro and promote the phagocytosis of peritoneal macrophage significantly in sepsis,which contribute to the protective effect on the sepsis mouse in the experimental sepsis induced by the intraperitoneal injection of E.coli.
3.In the experimental sepsis induced by LPS,FL may eliminate the toxic effect of free radical and enhance the proliferative response of lymphocytes,which contribute to the protective effect on the sepsis mouse.
研究金银花(Flos lonicerae,FL)提取物对小鼠T细胞行为和脓毒症的影响,探讨其对脓毒症小鼠的保护作用机制,为其在临床的应用提供参考数据和理论依据。
方法
1.体外结合流式细胞术利用荧光标记的单克隆抗体染色,检测FL对多克隆刺激剂刀豆蛋白A(Concanavalin A,Con A)刺激的小鼠淋巴细胞活化的影响;利用羧基荧光素二乙酸琥珀酰亚胺酯(CFDA-SE)染色分析FL对淋巴细胞增殖情况的影响;碘化丙锭(PI)染色分析FL对淋巴细胞周期分布的影响。
2.腹腔注射大肠杆菌(escherichia coli,E.coli)建立脓毒症模型,观察FL对小鼠生存状况的影响,分析各处理组小鼠胸腺、脾脏细胞总数,计算小鼠胸腺和脾脏指数,AnnexinV-FITC/PI双染检测FL对小鼠胸腺细胞凋亡的影响,使用CFDA-SE标记的大肠杆菌(E.Coli)结合流式细胞术检测FL对小鼠腹腔Mφ吞噬功能的影响。
3.利用腹腔注射LPS/D-半乳糖建立内毒素诱导脓毒症模型,采用MTT比色法检测FL对脓毒症小鼠淋巴细胞增殖的影响,通过H_2DCFDA染色检测外周血反应性氧族(reactive oxygen species,ROS)变化,利用H_2O_2测定试剂盒检测小鼠血清中H_2O_2的水平,通过血液生化和病理组织学检查,观察FL对小鼠肝功能的保护作用,结合体外试验,利用Griess反应试剂盒检测FL对脂多糖(LPS)激活的Mφ释放NO的影响。
结果
1.细胞活化分析结果表明,金银花提取物对小鼠淋巴细胞活化和增殖均有明显的抑制作用(P＜0.05),并且能明显阻止淋巴细胞进入S期和G2/M期。
2.大肠杆菌诱导的脓毒症小鼠胸腺和脾脏出现显著的萎缩现象;与此同时,药物治疗组小鼠胸腺和脾脏指数明显增加,结合体外实验,FL对DEX诱导的小鼠胸腺细胞凋亡都有显著的拮抗作用(P＜0.01);小鼠存活率亦显著提高(P＜0.01);腹腔巨噬细胞的吞噬功能明显增强。
3.利用内毒素诱导D-半乳糖致敏脓毒症模型,结果分析表明,与正常组相比,淋巴细胞对Con A诱导应答能力明显下降,外周血中性粒细胞ROS显著升高,血清中ALT、AST和H_2O_2水平亦增加,肝组织炎性细胞水平显著升高,肝脏出现病理损伤,而药物治疗组对增殖反应明显强于单纯模型组,外周血中性粒细胞ROS、血清中ALT、AST和H_2O_2水平均显著降低,肝组织炎性细胞水平减轻,Griess反应检测结果显示,FL明显抑制LPS激活巨噬细胞释放NO的量(P＜0.05)。
结论
1.FL能明显抑制小鼠淋巴细胞的体外活化和增殖,并能抑制淋巴细胞进入细胞分裂周期,提示FL对特异性细胞免疫有重要抑制作用,这可能是FL作为临床常用药物治疗免疫相关疾病的理论基础。
2.抑制胸腺和脾脏细胞凋亡和增强腹腔巨噬细胞吞噬功能可能FL对大肠杆菌所致脓毒症小鼠的保护机制。
3.清除氧自由基和提高脓毒症小鼠淋巴细胞免疫应答能力可能是FL对内毒素诱导的脓毒症小鼠的免疫保护机制。
Aim:
To investigate the effect of Flos lonicerae on mouse T lymphocytes behavior and sepsis,and elucidate the protective mechanism,in order to supply theory to clinical therapy.
Methods:
1.Lymphocytes isolated from lymphoid nodes of BalB/c mouse were stained with fluorescence conjugated monoclonal antibodies and flow cytometry analysis was employed to detect the effects of FL on the activation of mouse lymphocytes treated with Con A.The effects of FL on the proliferation rate of mouse lymphocytes were also analyzed by CFDA-SE staining.The distribution of the cell-cycle was analyzed by PI staining together with flow cytometry analysis.
2.For experiment sepsis,mouse was intraperitoneal injection(i.p.) of a bacterial suspension containing 10~8 live escherichia coli(E.coli) cells.We monitored the effect of FL on the mouse mortality rate in the experimental sepsis.The spleens and thymi were removed and cell yield was counted,and we compared spleen index and thymi index of different groups.Apoptosis of thymocytes DEX were detected by AnnexinV-FITC/PI staining together with flow cytometry.E.coli stained by CFDA-SE and flow cytometry was utilized to observe the effect of FL on macrophage phagocytosis.
3.For LPS-induced experimental sepsis,mice were given intraperitoneal injections of a mixture of LPS and D-galactosamine.The proliferation related index of mouse lymphocytes was analyzed with flow cytometry and MTT.Changes of reactive oxygen species(ROS) in blood were analyzed by H_2DCFDA staining together with flow cytometry.Serum ALT,AST levels and the pathological changes were obtained to study the protective of FL on liver function of septic mouse.Concentration of H_2O_2 in serum was measured by H_2O_2 kit.The effect of FL on NO production of the peritoneal macrophage in vitro was measured by Griess Reagent assay kit.
Results:
1.The results indicated that FL ihibited the activation of mouse lymphocytes(P＜0.05).CFDA-SE analysis also showed the proliferation of mouse lymphocytes were obviously inhibited by different concentrations of FL.Flow cytometry analysis of the cell-cycle distribution revealed that the cell number in G0/G1 was increased,while the cell number in S and G2/M was decreased as the concentration of FL increased.
2.The results demonstrated that septic mouse induced by the intraperitoneal injection of E.coli led to splenic atrophy.However,compared with septic mouse,FL could significantly increase thymus index and spleen index.FL could inhibit the apoptosis of thymus cells induced by Dex(P＜0.01).The survival rate of FL group was much higher than the sepsis group.FL promoted the phagocytosis.
3.Establishing sepsis model by intraperitoneal injections of LPS was to analysis the protective mechanism of FL on septic mouse.Compared with the control group,the lymphocytes proliferation reponses of septic mouse were reduced upon stimulation with Con A,and the levels of ALT,AST and H_2O_2 in serum and ROS in blood were increased significantly compared with the normal group,Meanwhile,the levels of inflammatory mediators were increased and morphological structures were damaged..But in the interference group FL could enhance proliferation of lymphocytes,and inhibit the levels of ALT,AST and H_2O_2 in serum and ROS in blood.Moreover,the levels of inflammatory mediators were significantly lower than septic group.FL could inhibit NO production of peritoneal macrophage induced by LPS with Griess Reagent assay kit.
Conclusion:
1.FL could significantly inhibit the activation and proliferation of mouse lymphocytes in vitro.Flow cytometry analysis of the cell-cycle distribution revealed that FL can inhibit the progression of mouse lymphocytes growing into S and G2/M phase. That indicated FL inhibited specific cellular immune response.These results supply theory to clinical therapy.
2.FL could inhibit atrophy in spleen and thymus induced by sepsis.FL could inhibit the apoptosis of thymus cells in vitro and promote the phagocytosis of peritoneal macrophage significantly in sepsis,which contribute to the protective effect on the sepsis mouse in the experimental sepsis induced by the intraperitoneal injection of E.coli.
3.In the experimental sepsis induced by LPS,FL may eliminate the toxic effect of free radical and enhance the proliferative response of lymphocytes,which contribute to the protective effect on the sepsis mouse.
引文
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[8]Chiffoleau E,Walsh P T,Turka L.Apoptosis and transplantation tolerance[J].Immunol Rev,2003,193:124-145.
[9]Ferguson T A,Stuart P M,Hemdon J M,Griffith T S.Apoptosis,tolerance,and regulatory T cells-old wine,new winekins[J],Immunol Rev,2003,193:111-123.
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[17] Woodside K J, Hu M, Meng T, Hunter G C, Sower L E. Differential effects of interleukin-2 blockade on apoptosis in naive and activated human lymphocytes [J]. Transplantation, 2003,75:1631-1635.
[1]Yah J J,Jung J S,Lee J E,Lee J,Huh S O.Therapeutic effects of lysophosphatidylcholine in experimental sepsis[J].Nat Med.2004,10(2):161-167.
[2]彭珊瑛,张弗盈,欧阳雪宇,刘洋,王文杰.银杏内酯B对血小板活化因子引起的巨噬细胞趋化及细胞骨架改变的影响[J].药学学报,2006,41(2):156-160.
[3]Efron P,Moldawer LL.Sepsis and the dendritic cell[J].Shock,2003,20(5):381-401.
[4]Bone RC.Sir Isaac Newton,sepsis,SIRS,and CARS[J].Crit Care Med,1996,24:1125-1128.
[5]Oberholzer A,Oberholer C,Moldawer LL.Sepsis syndromes:understanding the role of innate and acquired immunity[J].Shock,2001,16:83-96.
[6]Heidecke CD,Hensler T,Weighardt H,Zantl N,Wagner H,Siewert JR,Holzmann B.Selective defects of T lymphocyte function in patients with lethal intraabdominal infection[J].Am J Sung.1999,178:288-292.
[7]Hotchkiss R S,Tinsley K W,Swanson P E,Schmieg R E,Jia J H,Chang K C,Osborne D F,Freeman B D,Perren Cobb J,Buchman T G,Karl I E.Sepsis induced apoptosis causes progressive profound depletion of B and CD4+ T lymphocytes in humans[J].J Immunol,2001,166(1):6952-6963.
[8]Hotchkiss R S,Tinsley K W,Swanson P E,Grayson M H,Osborne D F,Wagner T H,Perren Cobb J,Coopersmith C,Karl I E.Depletion of dendritic cells,but not macrophages,in patients with sepsis[J].J Immunol,2002,168:2496-2500.
[9]Hotchkiss R S,Chang KC,Grayson MH,Tinsley K W,Dunne B S,Davis C G,Osborne D F,Karl I E.Adoptive transfer of apoptotic splenocytes worsens survival,whereas adoptive transfer of necrotic splenocytes improves survival in sepsis[M].Pro Natl AcadS-ci USA,2003,100:6724-6729.
[10]Le Tulzo Y,Pangault C,Gacouin A,Guilloux V,Tribut O,Amiot L,Tattevin P,Fauchet R.Early sirculating lymphocyte apoptosis in human septic shock is associated with poor outcome[J].Shock,2002,18(16):487-494.
[11]Fukuzuka K,Edwards C K,Clare-Salzler M,Copeland E M,Moldamei L L,Mozingo D W.Glucocorticoid-induced,caspase-dependent organ apoptosis early after bum injury[J].Am J Physiol Regul Integr Comp Physiol,2000,278(4):1005-1018.
[12] De Freitas I, Fernandez-Somoza M, Essenfeld-Sekler E, Cardier J E. Serum levels of the apoptosis-associated molecules, tumor necrisis factor-alpha/tumor necrosis factor type-1 receptor and Fas/FasL, in sepsis[J].Chest,2004,125(6):2238-2246.
[13] Calandra T, Echtenacher B, Roy DL, Pugin J. Protection from septic shock by neutralization of macrophage migration inhibitory factor[J]. Nat Med, 2000,6:164-170.
[14] Ayala A, Herdon CD, Lehman DL, Chaudry I H. Differential induction of apoptosis in lymphoid tissues during sepsis: variation in onset, frequency, and nature of the mediators[J]. Blood, 1996,87:4261-4275.
[15] Heidecke C D,Hensler T, Weighardt H, Zantl N, Wagner H, Siewert J R. Selective defects of T lymphocyte function in patients with lethal in intraabdominal infection[J].Am J Surg, 1999,178:288-292.
[16] Ayala A, Xu YX. Chung CS, Chaudry I H. Does Fas Ligand or endotoxin contribute to thymic apoptosis during polymicrobial sepsis [J]. Shock, 1999,11:211 -217.
[1]Dunican-AL,Lenenroth SL,Grutkoski P.TNF alpha-induced suppression of PMN apoptosis is mediated through interleukin-8 production[J].Shock,2000,14(3):284-289.
[2]Fujita S,Seino K,Sato K,Sato Y,Eizumi N.Regulatory dendritic cells act as regulators of acute lethal systemic inflammatory response[J].Blood,2006,107(9):3656-3664.
[3]Cohen J.The dectection and interpretation of endotoxemia[J].Inten Care Med 2000;26:51-56.
[4]Leist M,Gantner F,Bohlinger I,Tiegs G,Germann P G,Wendel A.Tumor necrosis Factor-induced hepatocyte apoptosis precedes liver failure in experimental murine of shock models[J].Am J Pathol,1995,146:1220-1234.
[5]Stachlewitz R F,Seabra V,Bradford B,Bradham C A,Rusyn I,Germolec D.Glycine and ufidine prevent D-galactosamine hepatotoxicity in the rat:role of kupffer cells[J].Hepatology,1999,29(3):737-745.
[6]许波,吴玉章.巨噬细胞诱导型一氧化氮合酶的表达调节机制[J].免疫学杂志,2002,18(3):156-159.
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[11]Sener G,Toklu H,Ercan F,Erkanli G.Protective effect of beta-glucan against oxidative organ injury in a rat model of sepsis[J].Int Immunopharmacol,2005,5(9):1387-1396.
[12] Zhang HB, Spapen H, Nguyen DN, Benlabed M, Buurman W A, Vincent J L. Protective effects of N-acetyl-L-cysteine in endotoxemia [J]. Am J Physiol, 1994, 266: H1746-H1754.
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[1]黄喜茹,刘伟娜,曹冬.金银花的化学成分 药理作用研究评价[J].中医药学刊,2005,23(3):418-419.
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[14]Werfel T,Boeker M,KappA.Rapid expression of the CD69 antigen on T cells and natural killer cells upon antigenic stimulation of peripheral blood mononuclear cell suspensions[J].Allergy, 1997,52(4):465469.
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[1]Yah J J,Jung J S,Lee J E,Lee J,Huh S O.Therapeutic effects of lysophosphatidylcholine in experimental sepsis[J].Nat Med.2004,10(2):161-167.
[2]彭珊瑛,张弗盈,欧阳雪宇,刘洋,王文杰.银杏内酯B对血小板活化因子引起的巨噬细胞趋化及细胞骨架改变的影响[J].药学学报,2006,41(2):156-160.
[3]Efron P,Moldawer LL.Sepsis and the dendritic cell[J].Shock,2003,20(5):381-401.
[4]Bone RC.Sir Isaac Newton,sepsis,SIRS,and CARS[J].Crit Care Med,1996,24:1125-1128.
[5]Oberholzer A,Oberholer C,Moldawer LL.Sepsis syndromes:understanding the role of innate and acquired immunity[J].Shock,2001,16:83-96.
[6]Heidecke CD,Hensler T,Weighardt H,Zantl N,Wagner H,Siewert JR,Holzmann B.Selective defects of T lymphocyte function in patients with lethal intraabdominal infection[J].Am J Sung.1999,178:288-292.
[7]Hotchkiss R S,Tinsley K W,Swanson P E,Schmieg R E,Jia J H,Chang K C,Osborne D F,Freeman B D,Perren Cobb J,Buchman T G,Karl I E.Sepsis induced apoptosis causes progressive profound depletion of B and CD4+ T lymphocytes in humans[J].J Immunol,2001,166(1):6952-6963.
[8]Hotchkiss R S,Tinsley K W,Swanson P E,Grayson M H,Osborne D F,Wagner T H,Perren Cobb J,Coopersmith C,Karl I E.Depletion of dendritic cells,but not macrophages,in patients with sepsis[J].J Immunol,2002,168:2496-2500.
[9]Hotchkiss R S,Chang KC,Grayson MH,Tinsley K W,Dunne B S,Davis C G,Osborne D F,Karl I E.Adoptive transfer of apoptotic splenocytes worsens survival,whereas adoptive transfer of necrotic splenocytes improves survival in sepsis[M].Pro Natl AcadS-ci USA,2003,100:6724-6729.
[10]Le Tulzo Y,Pangault C,Gacouin A,Guilloux V,Tribut O,Amiot L,Tattevin P,Fauchet R.Early sirculating lymphocyte apoptosis in human septic shock is associated with poor outcome[J].Shock,2002,18(16):487-494.
[11]Fukuzuka K,Edwards C K,Clare-Salzler M,Copeland E M,Moldamei L L,Mozingo D W.Glucocorticoid-induced,caspase-dependent organ apoptosis early after bum injury[J].Am J Physiol Regul Integr Comp Physiol,2000,278(4):1005-1018.
[12] De Freitas I, Fernandez-Somoza M, Essenfeld-Sekler E, Cardier J E. Serum levels of the apoptosis-associated molecules, tumor necrisis factor-alpha/tumor necrosis factor type-1 receptor and Fas/FasL, in sepsis[J].Chest,2004,125(6):2238-2246.
[13] Calandra T, Echtenacher B, Roy DL, Pugin J. Protection from septic shock by neutralization of macrophage migration inhibitory factor[J]. Nat Med, 2000,6:164-170.
[14] Ayala A, Herdon CD, Lehman DL, Chaudry I H. Differential induction of apoptosis in lymphoid tissues during sepsis: variation in onset, frequency, and nature of the mediators[J]. Blood, 1996,87:4261-4275.
[15] Heidecke C D,Hensler T, Weighardt H, Zantl N, Wagner H, Siewert J R. Selective defects of T lymphocyte function in patients with lethal in intraabdominal infection[J].Am J Surg, 1999,178:288-292.
[16] Ayala A, Xu YX. Chung CS, Chaudry I H. Does Fas Ligand or endotoxin contribute to thymic apoptosis during polymicrobial sepsis [J]. Shock, 1999,11:211 -217.
[1]Dunican-AL,Lenenroth SL,Grutkoski P.TNF alpha-induced suppression of PMN apoptosis is mediated through interleukin-8 production[J].Shock,2000,14(3):284-289.
[2]Fujita S,Seino K,Sato K,Sato Y,Eizumi N.Regulatory dendritic cells act as regulators of acute lethal systemic inflammatory response[J].Blood,2006,107(9):3656-3664.
[3]Cohen J.The dectection and interpretation of endotoxemia[J].Inten Care Med 2000;26:51-56.
[4]Leist M,Gantner F,Bohlinger I,Tiegs G,Germann P G,Wendel A.Tumor necrosis Factor-induced hepatocyte apoptosis precedes liver failure in experimental murine of shock models[J].Am J Pathol,1995,146:1220-1234.
[5]Stachlewitz R F,Seabra V,Bradford B,Bradham C A,Rusyn I,Germolec D.Glycine and ufidine prevent D-galactosamine hepatotoxicity in the rat:role of kupffer cells[J].Hepatology,1999,29(3):737-745.
[6]许波,吴玉章.巨噬细胞诱导型一氧化氮合酶的表达调节机制[J].免疫学杂志,2002,18(3):156-159.
[7]Ogilvie A C,Groeneveld A B,Straub J P,Thijs L G.Plasma lipid peroxides and antioxidants in human septic shock[J].lntersive Care Med,1991,17(1):40-44.
[8]Jabs CM,Neglen P,Eklof B.Breakdown of adenine nucleotides,formation of oxygen free radicals,and early markers of cellular injury in endotoxic shock[J].Eur J Surg,1995,161:147-155.
[9]Abello P A,Fidler S A,Bulkley G B,Buchman T G.Antioxidants modulate induction of programmed endothelial cell death(apoptosis) by endotoxin[J].Arcn Sury,1994,129:134-141
[10]Compton C N,Franko A P,Murray M T,Diebel L N,Dulchavsky S N.Signaling of apoptotic lung injury by lipid hydroperoxides[J].J Trauma,1998,44:783-788.
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