一氧化氮在肝损伤中作用的实验研究

材料与方法：１．动物模型的复制及分组：雄性ｗｉｓｔａｒ大鼠,随机分成六组。除正常对照组外,各组均给予１０％Ｄ－半乳糖胺（Ｄ－Ｇａ１Ｎ）０．６ｇ／ｋｇ及大肠杆菌Ｏ１１１Ｂ４脂多糖（ＬＰＳ）０．１ｍｇ／ｋｇ腹腔注射,造成大鼠急性肝损伤模型。并分别于给Ｄ－ＧａｌＮ前２０分钟及后６小时、１１小时,精氨酸（Ａｒｓ）组给子Ｌ－Ａｒｇ０．５ｇ／ｋｇ腹腔注射;Ａｒｇ一硝基精氨酸甲酯（ＮＡＭＥ）组给予Ｌ－Ａｒｇ０．５ｇ／ｋｇ加Ｌ一ＮＡＭＥ５０ｍｇ／ｋｇ;ＮＡＭＥ组给予Ｌ－ＮＡＭＥ１０ｍｇ／ｋｇ;地塞米松（Ｄｅｘ）…     

一氧化氮和自由基对大鼠急性肝损伤的作用

目的用硫代乙酰胺（ＴＡＡ）诱发大鼠急性肝损伤,观察肝损伤过程中一氧化氮与自由基的变化．方法实验Ⅰ：大鼠２４只分为４组,一组为正常组,其余３组为损伤组．ＴＡＡ６００ｍｇ／ｋｇｓｃ２４,４８,７２ｈ测定内毒素及ＮＯ３－／ＮＯ２－,ＡＬＴ,ＡＳＴ含量．取肝组织匀浆,测定蛋白含量．脂质过氧化物歧化酶（ＳＯＤ）及谷胱甘肽过氧化物酶（ＧＳＨＰＸ）活性,并观察肝组织学变化．实验Ⅱ：大鼠３２只分为４组,Ａ组为正常组,Ｂ,Ｃ,Ｄ组ＴＡＡ６００ｍｇ／ｋｇｓｃ;同时给予Ｂ组生理盐水０４ｍＬ／ｋｇ,Ｃ组７５％ＬＡｒｇ３００ｍｇ／ｋｇ,Ｄ组２５％ＬＮＮＡ１０ｍｇ／ｋｇ．２４ｈ后重复注射１次．２４ｈ后按实验Ⅰ取血、肝组织,测定有关指标．结果大鼠注射ＬＡｒｇ后,ＮＯ的合成增多,转氨酶及肝组织损伤程度明显降低,注射ＬＮＮＡ组大鼠肝损伤程度加重,肝组织自由基的测定表明,抑制肝损伤大鼠ＮＯ合成,肝组织ＬＰＯ含量增高而ＳＯＤ,ＧＳＰＨＸ活性降低,ＳＯＤ,ＧＳＨＰＸ协同作用可清除体内自由基．结论抑制ＮＯ的生物合成,自由基水平增高,从而加重了肝损伤     

丹参提取物（764—3）对不同类型肺动脉高压大鼠泡内动脉？…

目的 观察丹参提取物（７６４－３）对不同类型肺动脉高压大鼠肺腺泡内肌型（ＭＡ）、部分肌型（ＰＭＡ）和非肌型（ＮＭＡ）动脉百分比构在及ＭＡ中膜胶原含量的影响。方法 将雄性Ｗｉｓｔａｒ大鼠随机分为１２组。缺氧大鼠置于５１ｋＰａ低压舱内２或４周；野百合碱组（ＭＣＴ）大鼠以ＭＣＴ６０ｍｇ／ｋｇ一次腹腔注射，观察４周，给药大鼠：７６４－３ ２０或４０ｍｇ／ｋｇ皮下注射，每日１次；β氨基丙腈（ＢＡＰＮ）１５０     

拉西地平对高血压大鼠的降压作用

目的：观察新的钙拮抗剂拉西地平的降压作用。方法：急性降压实验：将５个月龄雄性卒中型自发性高血压大鼠（ＳＨＲｓｐ）随机分为Ａ、Ｂ、Ｃ（分别经管饲给药１次，拉西地平剂量１ｍｇ／ｋｇ，０．５ｍｇ／ｋｇ，０．２５ｍｇ／ｋｇ）、Ｄ（尼群地平１０ｍｇ／ｋｇ）、Ｅ（溶媒对照）组，每组１１只，于用药后１、３、５、９、１２、２４小时各测１次收缩压及心率（尾套法）。慢性降压实验：将３个月龄ＳＨＲｓｐ分为Ａ、Ｂ、Ｄ、Ｅ组（剂量同急性降压实验），每组１０只，治疗１４天，于治疗后第２、４、６、８、１０、１２、１４天各测１次收缩压及心率。结果：拉西地平１ｍｇ／ｋｇ和０．５ｍｇ／ｋｇ均可显著降低ＳＨＲｓｐ血压，降压作用分别持续１２～２４小时和９～１２小时，降压作用均强于尼群地平１０ｍｇ／ｋｇ，仅轻度增加了心率。结论：拉西地平对ＳＨＲｓｐ降压作用明确，持续时间长。     

吸入一氧化氮对急性损伤大鼠细胞间黏附分子—1表达的影响

近年的研究发现,一氧化氮(ＮＯ)具有抗黏附作用。本研究通过吸入ＮＯ治疗急性肺损伤(ＡＬＩ)大鼠,观察肺组织细胞间黏附分子(ＩＣＡＭ)1表达的变化,探讨ＮＯ的作用机制,为临床应用吸入ＮＯ治疗ＡＬＩ提供理论依据。一、材料与方法1动物分组与实验过程:雄性Ｗｉｓｔａｒ大鼠36只,体重250～380ｇ,随机分为6组,每组6只。对照组:静脉注射1ｍｌ/ｋｇ生理盐水;油酸模型组:静脉注射0.1ｍｌ/ｋｇ油酸(广州化玻仪器公司);内毒素模型组:静脉注射5ｍｇ/ｋｇ内毒素(ＬＰＳ)(Ｓｉｇｍａ公司)复制ＡＬＩ模型。致伤后1ｈ,ＮＯ对照组动物经鼻导管吸入含20ｐ…     

两种一氧化氮合酶抑制剂在内毒素肝损伤中的作用

材料与方法：内毒素（ＬＰ）（Ｄｉｆｉｃｏ,美国）硝基左旋精氨酸（Ｌ－ＮＮＡ）（Ｓｉｇｍａ）,Ｓ－硫酸甲基异硫脲素（ＳＭＴ,Ｓｉｍａ）,丙氨酸转氨酶（ＡＬＴ）测定试剂盒（上海荣胜生物制剂厂）。Ｗｉｓｔａｒ大鼠（重庆医科大学实验动物中心）雄性,体重２００～３００ｇ。动物分为４组,第１组腹腔注射灭菌生理盐水１ｍｌ;第２组腹腔注射ＬＰＳ（４ｍｇ／ｋｇ）;第３组在腹腔注射ＬＰＳ（４ｍｇ／ｋｇ）前１小时,动物先腹腔注射Ｌ－ＮＮＡ（１００ｍｇ／ｋｇ）;第４组注射ＬＰＳ（４ｍｇ／ｋｇ）前２小时,腹腔注射ＳＭＴ（…     

榆林病区水黄腐酸对大鼠的急性毒性实验报告

报告了榆林大骨节病新发户水黄腐酸（ＦＡ）对大鼠急性毒性实验结果。以４００ｍｇ·ＦＡ／ｋｇＢＷ一天灌胃于组Ⅲ的４只鼠观察了７天，以３００ｍｇ·ＦＡ／ｋｇ ＢＷ·ｄ灌胃于组Ⅱ的５只鼠，其中１只连续３天，１只４天，１只６天，余２只连续７天。全部动物于实验第８天取材处死。实验结果：未见两组大鼠ＦＡ致死的鼠，未见两组大鼠主要生命器官和骨软骨病理形态变化。组Ⅲ鼠摄ＦＡ量是病区儿童每公斤体重等效剂量的６８００倍     

内源性一氧化氮对犬急性缺氧性肺动脉高压的影响

利用一氧化氮合成酶抑制剂——Ｎ￣Ｇ－硝基－Ｌ－精氨酸甲酯（Ｌ－ＮＡＭＥ）观察内源性一氧化氮对犬急性缺氧性肺血管收缩的影响。Ｌ－ＮＡＭＥ（１、５、１５ｍｇ／ｋｇ）可显著增加缺氧时肺动脉平均压（ｍＰＡＰ）和肺血管阻力（ＰＶＲ），其作用维持９０～１８０分钟。与用药前缺氧时比较，ｍＰＡＰ最大可分别升高１．１±０．２、１．５±０．３、１．６±０．４ｋＰａ（１ｋＰａ＝７．５ｍｍＨｇ），而ＰＶＲ最大可分别升高５８．４±１５．６、９９．３±２８．８、７８±４．０ｋＰａ·ｓ／Ｌ，Ｌ－ＮＡＭＥ１５ｍｇ／ｋｇ升ｍＰＡＰ的作用明显强于１ｍｇ／ｋｇ组。Ｌ－精氨酸（０．５ｇ／ｋｇ）可逆转Ｌ－ＮＡＭＥ（５ｍｇ／ｋｇ）增强缺氧性肺血管收缩的作用。此外，Ｌ－ＮＡＭＥ也可明显增加缺氧时股动脉平均压和全身血管阻力，同时减少心输出量，减慢心率，结果提示内源性一氧化氮可能具有抑制急性缺氧性肺血管收缩的作用。     

山莨菪碱对实验性急性肝损伤的保护效果

观察山莨菪碱对实验性肝损伤的作用，并对其机理作初步探讨。资料与方法：体重２１０～２６０ｇ雄性Ｗｉｓｔａｒ大鼠，随机分成３组，每组６只。用Ｄ－半乳糖胺０６ｇ／ｋｇ加内毒素脂多糖０１ｍｇ／ｋｇ制成急性肝损伤模型，山莨菪碱（Ａｎｉ）组动物在上述处理的前...     

靶向伯氨喹抗约氏疟原虫红外期作用的初步研究

用半乳糖基新糖白蛋白（Ｇ－ＮＧＡ）包裹伯氨喹（ＰＱ）制成的靶向伯氨喹（ＴＰＱ）静脉注射（ｉｖ）感染约氏疟原虫子孢子２ｈ的Ｗｉｓｔａｒ大鼠，４２ｈ时，２０ｍｇ／ｋｇ和１０ｍｇ／ｋｇＴＰＱ组子孢子发育率（ＳＰＶ）分别为１．２％和２．８％，仅分别为对照组的６％左右和１５％，约有９０％红外期（ＥＥＦ）呈现退变或发育受到明显抑制；２０ｍｇ／ｋｇＰＱ组ＳＰＶ为５．０％，为对照组的２６％，半数以上ＥＥＦ均呈退变或发育受到抑制。昆明株小鼠ｉｖ约氏疟原虫子孢子后２ｈ分别注射药物，ＴＰＱ１０ｍｇ／ｋｇ组和ＰＱ２０ｍｇ／ｋｇ组原虫血症前期较对照组明显后延，尤其是ＴＰＱ１０ｍｇ／ｋｇ组中２只鼠均未出现原虫血症。结果提示ＴＰＱ抗ＥＥＦ药效明显优于ＰＱ。     

急性肺损伤大鼠多形核白细胞L选择素变化及其在肺内扣押中的作用

目的 探讨急性肺损伤(ALI)大鼠循环血多形核白细胞(PMN)L选择素表达的变化及其于肺内扣押中的作用.方法 通过静脉注射内毒素(5 mg/kg)复制大鼠ALI模型,56只大鼠随机分为7组,每组8只.分别为(1)内毒素5 min组;(2)内毒素15 min组;(3)内毒素30 min组;(4)内毒素60 min组;(5)fucodin干预5 min组;(6)Fucodin干预15 min组静脉注射fucodin 5 mg/kg后,立即静脉注射内毒素5 mg/kg;(7)正常对照组静脉注射等量生理盐水替代内毒素.用免疫荧光间接法和流式细胞仪检测大鼠ALI过程中PMN L选择素蛋白表达的变化;用髓过氧化酶(MPO)分析法及组织学检查定量ALI过程中PMN于肺内的扣押量.结果 (1)PMN L 选择素的表达于静脉注射内毒素后5 min为7.8±1.6,与对照组(10.5±2.1)比较差异有显著性(P<0.05),静脉注射内毒素后15 min(2.9±0.5)、30 min(3.5±0.7)和60 min(4.9±0.7)与对照组比较差异更具显著性(P<0.01).(2)大鼠肺组织MPO活力于静脉注射内毒素后5 min [(0.359±0.074) U/mg肺组织]、15 min [(0.490±0.069) U/mg肺组织]、30 min [(0.565±0.111 ) U/mg肺组织]、60 min [(0.710±0.112) U/mg肺组织]与对照组[(0.069±0.011) U/mg肺组织]比较差异有显著性(P<0.01);fucodin干预5 min组[(0.391±0.071)U/mg肺组织]和对应时相点的内毒素组[(0.359±0.074) U/mg肺组织]比较差异无显著性,fucodin干预15 min组[(0.396±0.061) U/mg肺组织]和对应时相点的内毒素组[(0.490±0.069) U/mg肺组织]比较差异有显著性(P<0.05).结论 (1)正常状态下L选择素在PMN表面呈结构性表达,内毒素致伤后PMN L选择素表达迅速减少,伤后15 min时最低,其后呈回升趋势.(2)内毒素性ALI大鼠PMN肺内的早期扣押可能是L选择素非依赖性的,但L选择素对维持肺PMN的持续扣押仍然是重要的.     

Modulation by pentobarbital of neutrophil responses to inhaled E. coli endotoxin in sheep: role of lung epithelium.

Neutrophils (PMNs) are implicated in the pathogenesis of acute respiratory distress syndrome (ARDS). The role of the epithelium in the modulation of PMN migration within the lungs was examined. Epithelial integrity and PMN concentrations in the lung air spaces and lymph were measured in sheep anaesthetized with either halothane (1-2.5%) or intravenous pentobarbital (12+/-4 mg x kg(-1) x h(-1)). Ventilation with an aerosol containing 25 mg Escherichia Coli endotoxin (lipopolysaccharide; LPS) effected neutrophil recruitment to the air spaces. Lymphatic clearance of aerosolized 99mTc-DTPA provided an index of epithelial integrity. Three hours after the deposition of LPS, the lung lining fluid of sheep anaesthetized with halothane (n=7) had 4.9+/-3.2x10(6) PMN.mL(-1), but the lung lymph had almost no PMNs (3+/-8%). Sheep anaesthetized with pentobarbital (n=6) had fewer PMNs in the air spaces (2.4+/-1.2X10(6) mL(-1)) and more PMNs in the lung lymph (30+/-20%). Control sheep (n=5) that received no LPS had almost no PMNs in the airspaces or lung lymph, regardless of the anaesthesia. Three additional sheep that remained awake after receiving LPS also had no PMNs in the lung lymph. The PMN fraction in the lung lymph correlated well with the extra-alveolar epithelial permeability measured by lymphatic clearance of aerosolized diethylenetriamine penta-acetic acid (r=0.81, p<0.001). Aerosolized lipopolysaccharide recruits neutrophils into the lungs of sheep, but they appear to remain in the airspaces unless extra-alveolar permeability is increased by agents such as pentobarbital.     

The influence of iron deficiency on erythrocyte deformability.

The influence of iron deficiency on erythrocyte deformability is controversial. The present study was designed to analyse cell deformability in 14 patients with iron deficiency and controls in a comprehensive way by three different methods, namely erythrocyte filtration, erythrocyte elongation, and measurement of membrane elasticity. Suspensions of washed erythrocytes (haematocrit 0.10) free of leucocytes were used. Erythrocyte deformability measured by filtration was increased by iron deficiency: The relative filtration resistance of a cell in a 3 microns pore was 26.5 +/- 6.9 and 75.8 +/- 23.8 in iron deficiency and controls, respectively (P less than 0.0001). In 5 microns pores the values were 2.80 +/- 1.23 and 3.46 +/- 0.51 (not significant); when the red cell number/volume was adjusted to that in control samples, the value for iron deficiency became significantly lower than in controls (2.32 +/- 0.60, P less than 0.0001). Erythrocyte elongation by centrifugation was unaffected (ratio length/width 1.66 +/- 0.11 and 1.60 +/- 0.10 in iron deficiency and controls, respectively). Membrane elasticity, as assessed by a filter aspiration technique, was also unchanged (membrane elastic modulus 3.94 +/- 0.31 and 3.94 +/- 0.37 x 10(-3) dyn/cm, respectively). It is concluded that iron deficiency does not affect erythrocyte membrane elasticity and that the deformability of whole cells is not impaired, but improved under certain conditions such as the passage of 3 microns pores because of microcytosis with preserved surface/volume ratio. These results are in contrast to earlier studies and they have pathophysiological and clinical implications.     

冠状动脉内应用山莨菪碱对急性心肌梗死介入治疗后无再流及心室功能和收缩同步性的影响

目的探讨冠状动脉内应用山莨菪碱对急性心肌梗死介入治疗（AMI-PCI）后无再流患者的逆转作用并评价其对患者局部、整体心室功能和收缩同步性的影响。方法自2003年1月至2006年2月首发急性前壁心肌梗死并于12h内行急诊PCI的患者136例,根据心肌灌注分级方法（myocardial blush grade,MBG）确认无再流患者（MBG0-1级）47例（男36例,女11例）,平均年龄（63．23±11．24）岁,随机分为两组：A组（山莨菪碱组,24例）和B组（对照组23例）,A组于PCI后即刻由指引导管冠状动脉内注射山莨菪碱1000斗g／次,余治疗同B组。于PCI后即行左心室造影,测定心室容积、压力参数和室壁运动积分（wall motion score,WMS）;AMI后1周时行平衡法核素心室造影,测定左室整体和局部收缩功能、舒张功能和收缩同步性参数;AMI后6个月随访时重复行心室造影和核素心室造影检查测定上述参数,同时随访并记录术后6个月内主要不良心脏事件（MACE）的发生率。结果（I）A组患者在冠状动脉内应用山莨菪碱1000μg／次,平均（2．53±0．34）次后MBG由（0．74±0．32）级增加到用药后的（2．33±0．28）级。（2）AMI-PCI后6个月随访时,A组左室收缩末容积指数、左心室舒张末期容积指数、WMS和左室舒张末期压均较B组明显降低[（40．53±8．12）mL／m^2比（50．32±8．26）mL／m^2,（80．13±9．74）ml／m^2比（87．17±10．25）mL／m^2,（8．24±1．31）比（10．23±1．82）,（13．36±4．21）mmHg（1mmHg=0．133kPa）比（16．38±3．21）mmHg,P均〈0．05];核素心室造影参数比较,A组左室射血分数、峰射血率和峰充盈率等参数均较B组明显增加I（44．02±5．86）％比（38．52±5．18）％,（1．86±0．09）EDV／s比（1.61±0．09）EDV／s,（2．19±0．32）EDV／s比（1.78±0．17）EDV／s,P均〈0．05]。（3）A组AMI-PCI后6个月左室局部射血分数（LrEF）2-LrEF8均分别较B组增加13．96％、25．02％、30．36％、22．86％、27．67％、22．07％和18．71％（P均〈0．05）.（4）相位分析示A组左室收缩同步性参数相角程、半高宽和峰相位标准差亦均低于B组[（46．04±8．93）°比（53．19±16．62）°,P〈0．05;（23．02±6．27）°比（25．02±5．31）°,P〉0．05;（7．92±4．12）°比（11．76±4．11）°,P〈0．05]。（5）在6个月随访期内,A组MACE发生率明显低于B组。结论冠状动脉内注射山莨菪碱可明显逆转AMI-PCI后无再流现象,改善无再流患者的心室功能和收缩同步性,降低MACE发生率。     

Effects of salvianolic acids on erythrocyte deformability in oleic acid induced acute lung injury in rabbits

The present study was to investigate the protective effects of salvianolic acids (SA) on deformability of red blood cells (RBCs) and its mechanism during the development of acute lung injury (ALI) induced by oleic acid (OA) in rabbits. 32 rabbits were randomized into four groups, normal control group, OA-treated group (0.15 ml/kg), SA-treated group and OA+SA treated group. The blood samples were collected at 0, 10, 30, 60, 90, 120 and 180 min after OA injection. The RBC deformation index, Orientation index and small deformation index were measured by ektacytometry. The concentration of malondialdehyde (MDA) in RBCs was detected by the assay kit. Meanwhile, the pulmonary pathological examination and the blood gas analysis were also performed. The results showed that the deformation index, orientation index and small deformation index decreased during the early phase of ALI, while the concentration of MDA in RBCs increased during the course. Pre-treatment with SA increased the deformability and orientability of RBC significantly and decreased the concentration of MDA in RBCs compared with OA group. Meanwhile, the hypoxia and pulmonary pathological damage were much improved. These results suggest that there were erythrocyte deformability changes in the early phase of ALI. SA has the protective effects on erythrocyte deformability during the development of ALI induced by OA, which might be due to its antioxidant effect. These results are valid in rabbits and in a model of ARDS, it would be interesting to see the effects of SA in patients.     

Red blood cell deformability in iron deficiency anaemia

In patients with iron deficiency anaemia (IDA) it has been suggested that the shortened erythrocyte lifespan may be in part due to decreased erythrocyte deformability. In order to know whether erythrocyte deformability is decreased in IDA patients, we have determined the erythrocyte Elongation Index (EI) by means of ektacytometric techniques (Rheodyn SSD, Myrenne Gmbh, Germany), in 50 IDA patients and 100 well age and sex matched healthy controls. At the three shear stresses tested, 12, 30 and 60 Pa, IDA patients show statistically lower EI than controls (37.4+/-6.7 vs 48.6+/-2.9; 45.0+/-6.0 vs 54.5+/-2.8; 48.7+/-5.8 vs 57.0+/-2.9 mPa.s, respectively; p<0.001). A statistically significant correlation was found between EI at 12, 30, and 60 Pa and the hematimetric indices (MCV, MCH and MCHC), suggesting that the alteration in surface/volume ratio (shape) which characterizes this kind of microcytic hypocromic anaemia, accounts in part for the decreased EI. Rheodyn SSD, as an ektacytometric technique, is very sensitive to alterations in red blood cell geometry, for what seems to be a useful tool for detecting diminished erythrocyte deformability in IDA patients.     

粒细胞集落刺激因子在急性肺损伤发病机制中作用的初步 …

目的：探讨粒细胞集落刺激因子（Ｇ－ＣＳＦ）在急性肺损伤（ＡＬＩ）发病过程中的作用。方法通过大鼠腹腔内注射内毒素建立ＡＬＩ模型。建立３０只大鼠分为５组：正常对照组及内毒素注射后２ｈ、４ｈ、６ｈ、８ｈ４个时相组,采用逆转录多聚酶链反应（ＲＴ－ＰＣＲ）的方法检测肺组织内Ｇ－ＣＳＦ ｍＲＮＡ表达水平及相关指标。结果内毒素腹腔注射２ｈ组肺内Ｇ－ＣＳＦ ｍＲＮＡ表达明显高于正常对照组,４ｈ组达到最高值,２ｈ组     

Erythrocyte deformability in obesity measured by ektacytometric techniques

Erythrocyte deformability (ED) has been scarcely evaluated in obese patients without other concomitant cardiovascular risk factors and contradictory results have been published regarding the influence of plasma lipids on the erythrocyte membrane lipid composition and insulin resistance on this rheological parameter. In 67 severe or morbid obese patients without other cardiovascular risk factors (51 women and 11 men, aged 34+/-11 years) and in 67 controls (45 women and 22 men, aged 32+/-10 years), ED has been determined by ektacytometric techniques in a Rheodyn SSD, the elongation index (EI) being measured at 12, 30 and 60 Pa, along with plasma lipids, red blood cell membrane lipids (cholesterol and phospholipids) and insulin resistance indexes in basal conditions and after a three month diet period. No significant differences were obtained in the EI between obese patients and the control group at any of the shear stresses tested (P>0.05). The cholesterol and phospholipid content of the red blood cell membrane did not significantly differ between cases and controls (P>0.05). Obese patients with metabolic syndrome showed lower EI at 30 and 60 Pa than those without metabolic syndrome (P=0.014 and P=0.031 respectively). Weight loss was not accompanied by any changes in these rheological parameters. Obesity itself does not seem to modify ED. However, metabolic syndrome seems to decrease ED, possibly through insulin resistance.     

Increased infection mortality and decreased neutrophil migration due to a component of an artificial blood substitute

We previously showed that an artificial blood substitute containing perfluorocarbons, Fluosol-DA, inhibited both neutrophil migration and adherence, due to its detergent component, Pluronic F-68. The purpose of the studies we report here was to determine if Fluosol or Pluronic might also reduce in vivo neutrophil migration and impair host resistance to bacterial infection. We studied in vivo PMN migration by injecting mice intraperitoneally (IP) with glycogen, followed by intravenous (IV) infusion of saline, Fluosol, or Pluronic. Peritoneal lavage after eight hours showed a significant decrease in the accumulation of PMN in lavage fluids of animals given either Fluosol or Pluronic (control--.19 +/- .03 X 10(6) PMN/mL, glycogen--1.35 +/- .14; glycogen/Fluosol--0.63 +/- .12; glycogen/Pluronic--0.69 +/- .07). We ascertained the effect of Fluosol and Pluronic on infection mortality by injecting mice IV with saline, Fluosol, or Pluronic, followed by a quantity of E coli (0.6 X 10(7] IP shown in preliminary studies to kill 20% to 50% of the mice in 24 hours. The 24-hour mortality was 14/45- saline, 24/32-Fluosol (chi 2 = 17.1; P less than .001) and 17/23 - Pluronic (chi = 11.2; P less than .001). Neither Fluosol nor Pluronic caused mortality without E coli. The increase in infection mortality occurred when Fluosol was given either two hours before, or simultaneously with E coli, but only with the simultaneous administration of bacteria and Pluronic. Pluronic did not alter reticuloendothelial system (RES) clearance function. These studies indicate that, in an animal model, Fluosol-DA, due to its detergent component Pluronic F-68, impaired neutrophil delivery to an inflammatory locus, and resulted in an increased rate of infection mortality. Since Pluronic did not result in RES blockade, but did impair the delivery of PMN to an inflammatory locus, our results suggest that the latter effect is responsible for the increase in infection mortality.     

Circulating neutrophil priming and systemic inflammation in limb ischaemia-reperfusion injury.

BACKGROUND: Recruitment and activation of neutrophils is a key step in the development of local and systemic injury in lower limb ischaemia-reperfusion. We hypothesis that increased circulating neutrophil priming is responsible for systemic inflammation. METHODS: Anaesthetised ventilated swine (n = 6 per group) underwent mid-line laparotomy and were randomised to control group or bilateral external iliac artery occlusion for two hours followed by two and a half hours reperfusion (I/R group). Using luminol, respiratory burst activity was assayed with a BioOrbit Luminometer to detect whole blood chemiluminescence (CL) by stimulation with phorbol 1,2-myristate 1,3-acetate (PMA) in the absence or presence of tumour necrosis factor (TNF) respectively. PMN priming is expressed as the ratio of whole blood CL in the presence of TNF to that without. We measured plasma interleukin(IL)-6 and tumour necrosis factor alpha by bioassay as a measure of systemic inflammation. The alveolar-arterial (A-a) gradient was measured using the formula [(A-a)gradient = fraction inspired O2 x 710-(arterial pCO2/0.8)-arterial pO2], it is a measure of lung function, a large gradient being indicative of impaired oxygen transport and hence lung injury. RESULTS: Lower limb I/R caused significantly greater PMN priming, 0.83 +/- 0.14, compared to control group, 0.22 +/- 0.04, (p < 0.001). Plasma IL-6, a reliable indicator of systemic inflammation, was significantly increased in I/R group after two and a half hours of reperfusion, 1295.0 (833.9-2073.0) pg/L, compared to control, 382.9 (367.4-568.3) pg/L, (p < 0.005). Plasma tumour necrosis factor alpha was significantly elevated after one hour of reperfusion in the I/R group, 86.8 (48.7-106.6) pg/ml, compared to the control group, 32.7 (0.9-42.8) pg/ml, (p < 0.01). (A-a) gradient was significantly increased after IRI, 407.97 +/- 53.13, compared to the control, 183.19 +/- 45.75, (p < 0.005). Mean pulmonary artery pressure was significantly greater after IRI, 38.80 +/- 4.87 mmHg, compared to control, 27.86 +/- 1.92 mmHg, (p < 0.005). Data represents mean +/- standard error mean or median (interquartile range), statistical comparisons using one-way Anova with Student's "t"-test and Kruskall-Wallis Anova with the Mann-Whitney U test. CONCLUSIONS: Priming of neutrophils increases their circulating respiratory burst activity and ability to induce tissue injury. Systemic PMN priming during hind limb ischaemia-reperfusion injury is associated with the systemic inflammatory response syndrome.