内毒素对体外培养大鼠肺微血管内皮细胞的影响

目的：探讨内毒素（ＬＰＳ）对体外培养鼠微血管内皮细胞（ＭＥＣ）的影响。方法：应用体外培养的鼠肺ＭＥＣ,观察ＬＰＳ对其分泌功能及ｓＩＣＡＭ－１和ｓＥ－ｓｅｌ表达的影响。结果：（１）在ＬＰＳ作用下,ＥＴ－１和ＮＯ６０分钟高于对照组,１８０分钟达高峰,高水平状态持续到２４０分钟,３６０分钟有下降趋势,ＬＰＳ高峰浓度为１ｍｇ／Ｌ。（２）ｓＩＣＡＭ－１在ＬＰＳ作用后６０分钟表达迅速上调,呈时间、剂量依赖性,     

创伤后多脏器功能衰竭患者细胞粘附分子和补体激活成分水平的变化及意义

目的：探讨创伤后多器官功能衰竭（ＭＯＦ）患者血浆粘附分子和补体活化成分水平的变化及意义。方法：采用酶联免疫吸附法（ＥＬＩＳＡ）检测３６例创伤后ＭＯＦ患者及３１例创伤患者和３５例健康人外周血白细胞ＣＤ１８的变化、血浆可溶性细胞间粘附分子－１（ｓＩＣＡＭ－１）,可溶性血管细胞间粘附分子－１（ｓＶＣＡＭ－１）和血浆补体活化片段（ｓＣ５ｂ－９）浓度的变化。结果：ＭＯＦ患者白细胞ＣＤ１８的表达、ｓＩＣＡＭ－     

肾小球肾炎患者WBC流变性和CD18表达及血清sICAM—1浓度测定及其意义

探讨ＷＢＣ流变性和细胞粘附分子（ＣＡＭｓ）与肾小球肾炎病情变化的关系。方法采用红细胞变形能力测定仪、体外。栓血小板粘附两用仪及酶联免疫吸附法（ＥＬＩＳＡ）,分别检测了４０例急性肾小球肾炎患者、４２例慢性肾小球肾炎患者和４５例健康人外周血白细胞滤过指数（ＬＥＩ）、白细胞粘附率（ＬＡＲ）、白细胞ＣＤ１８表达及血清可溶性细胞间粘附分子一１（ＳＩＣＡＭ－１）浓度的变化。结果肾小球肾炎患者ＬＦＩ,ＬＡＲｔ,白细胞ＣＤ１８表达和ｓＩＣＡＭ－１浓度均明显增高,与对照组比较差异有极显著性（Ｐ均＜０．００１）,且慢性肾小球肾炎患者各指标增高较急性肾小球肾炎患者更明显（Ｐ均＜０．００１）。结论白细胞流变性异常及ＣＤ１８表达、血清ｓＩＣＡＭ－１浓度增高与肾小球肾炎病情变化有密切关系。     

自身免疫病患者血清胞间粘附分子检测的意义探讨

目的 了解进行性硬皮病（ＰＳＳ）、类风湿性关节炎（ＲＡ）、系统性红斑狼疮（ＳＬＥ）血清可溶性细胞间粘附分子（ｓＩＣＡＭ）１和３的水平及与病情的关系。方法 用进口酶联免吸附法（ＥＬＩＳＡ）试剂盒分别检测活动期ＰＳＳ、ＲＡ、ＳＬＥ患者与正常人血清ｓＩＣＡＭ－１以及ＰＳＳ、ＲＡ患者、正常人血清ｓＩＣＡＭ－３的含量，比较其水平的差异。结果 正常对照组（２０例）ｓＩＣＡＭ－１和ｓＩＣＡＭ－３水平分别为２０２     

ICAM—1／LFA—1和HLA—DR在蕈样肉芽肿中的表达及其意义

目的：为探讨粘附分子在蕈样肉芽肿（ＭＦ）病理形成机制中的作用。方法：采用免疫组化技术，检测了１６例ＭＦ皮损部位ＩＣＡＭ－１／ＬＦＡ－１和ＨＬＡ－ＤＲ的表达。结果：ＩＡＣＭ－１／ＬＦＡ－１和ＨＬＡ－ＤＲ在ＭＦ表皮和／或毛囊上皮角朊细胞 不同程度的表达，其中ＩＣＡＭ－１的表达与ＭＦ亲表皮性明显相关。结论Ｌ：ＩＣＡＭ／１ＬＦＡ－１在ＭＦ亲表皮性形成过程中可能起着重要作用。     

山莨菪碱和蜕皮激素对内毒素致体外内皮细胞能量代谢的保护作用

目的：探讨山莨菪碱（６５４－２）和蜕皮激素９ＥＤＳ）对内毒素（ＬＰＳ）致伤培养脐静脉内皮细胞（ＨＵＶＥＣｓ）能量代谢的保护作用。方法：采用反相市郊和液相色谱分析法测定ＬＰＳ损伤后培养ＨＵＶＥＣｓ ＡＴＰ、ＡＤＰ、ＡＭＰ及能量负荷（ＥＣ）的变化以及６５４－２、ＥＤＳ的保护作用。结果：ＬＰＳ与内皮细胞孵育后低浓度、早期内皮细胞被激活,ＬＰＳ同一浓度刺激后,随刺激时间延长,ＨＵＶＥＣｓ ＥＣ降低,与正常     

白血病和骨髓增生异常综合征患者红细胞酶及同工酶谱的改变

目的：调查白血病和骨髓增生异常综合征（ＭＤＳ）患者红细胞酶和同工酶谱改变的患病率，并研究其临床意义。方法：采用ＩＣＳＨ介绍的方法及ＰＡＧＥ电泳测定红细胞酶活性及ＰＫ和ＡＤＡ同工酶，采用ＦＰＬＣ系统纯化ＡＤＡ同工酶，分析酶蛋白的生化性质。结果：患病率：在２２９例白血病和ＭＤＳ患者中，Ｇ６ＰＤ缺乏为４３．１％，ＰＫ缺乏为２７．８％。ＭＤＳ患者中的酶缺乏患病率：ＰＫ缺乏为５０．０％，Ｇ６ＰＤ缺乏为４８．０％，ＰＮＰ缺乏为３９．１％；酶过多患病率：ＡＤＡ过多为６９．６％，ＥＮＯＬ过多为４０．０％，ＡＬＤ过多为３０．４％。慢性粒细胞白血病（ＣＭＬ）的ＰＫ活性增高，而ＰＫ同工酶型均正常。急性白血病患者采用抗肿瘤抗生素后Ｇ６ＰＤ和６ＰＧＤ活性减低（Ｐ＜０．０５）。ＭＤＳ红细胞酶缺陷显著多于ＡＤＡ２同工酶表达见于ＡＭＬ（Ｍ４和Ｍ５）的单个核细胞及慢性粒－单核细胞白血病（ＣＭＭＬ）的红细胞，但ＡＬＬ不表达（Ｐ＜０．００１）。ＡＤＡ同工酶纯化分析示正常结构。结论：红细胞酶的检测可作为ＣＭＬ的预后参数，作为急性白血病抗肿瘤药物耐药预示指标，有助于ＭＤＳ－ＲＡ和再障、ＣＭＬ与ＣＭＭＬ的鉴别诊断。     

内毒素条件下中性粒细胞／血管内皮细胞膜表面CD18／ICA…

应用细胞酶联免疫吸附测定法（ＲＬＩＳＡ）测定内纱刺激条件下，中性粒细胞（ＰＭＮ）和增益脐静脉内皮细胞（ＶＥＣ）膜表面主要粘附分子的表达，前者检测ＣＤ１８，后者检测作为ＣＤ１８配体的细胞间粘附分子－－１（ＩＣＡＭ－１）。结果表明：内毒素可迅速导致ＰＭＮ膜表面ＣＤ１８的表达，半小时内达表达高峰，增另刺激时间不能持续其表达水平，ＶＥＣ在内毒素刺激诱导３ｈ后，ＩＣＡＭ－１的表达在一定基础表达水平上开始持续     

冠心病患者血清可溶性细胞间粘附分子—1和血管细胞粘附分子—1的 …

检测各型冠心病患者循环中可溶性细胞间粘附分子－１和血管细胞粘附分子－１水平并分析它们与冠心病活动性之间的关系。方法：用ＥＬＩＳＡ法测定稳定型心绞痛患者者,不稳定型心绞痛患者,急性心肌梗塞患者和健康体检者血清ｓＩＣＡＭ－１和ｓＶＣＡＭ－１浓度。     

急性髓系白血病细胞侵袭能力与粘附分子表达关系探讨

目的：探讨白血病细胞浸润及转移与粘附分子表达的关系。方法：用免疫组织化学ＡＰＡＡＰ、免疫印迹方法研究５０例急性髓系白血病（ＡＭＬ）骨髓和（或）外周血白血病细胞粘附分子ＶＬＡ４（ＣＤ４９ｄ）、ＬＦＡ１（ＣＤ１１ａ）的表达。结果：发现ＡＭＬ浸润组ＣＤ４９ｄ、ＣＤ１１ａ表达较非浸润组显著增高（Ｐ＜０．００５）。结论：ＡＭＬ白血病细胞可能通过ＣＤ４９ｄ／ＶＣＡＭ－１，ＣＤ１１ａ／ＩＣＡＭ－１粘附机制与血管内皮细胞粘附而浸润组织。外周血白血病细胞与骨髓白血病细胞ＣＤ４９ｄ、ＣＤ１１ａ表达无显著差别，说明骨髓白血病细胞ＣＤ４９ｄ、ＣＤ１１ａ表达高低不是其从骨髓释出的唯一因素。     

山莨菪碱对脑微血管内皮细胞缺氧-复氧时细胞间黏附分子-1的影响

目的：探讨山莨菪碱（Ani）对缺氧－复氧时脑微血管内皮细胞间黏附分子－1（ICAM－1）表达的影响。方法：采用流式细胞仪检测脑微血管内皮细胞在缺氧、缺氧－复氧及加入Ani后ICAM－1的阳性细胞百分率。结果：缺氧12h，脑微血管内皮细胞ICAM－1的表达与常氧时差异无显著意义，缺氧12h再给氧12h，ICAM－1的表达明显增加，加入Ani后，ICAM－1的阳性细胞百分率明显降低。结论：Ani可减少缺氧－复氧时脑微血管内皮细胞ICAM－1的表达。     

姜黄素对缺氧再给氧大鼠心脏微血管内皮细胞ICAM-1表达的影响

目的：研究姜黄素对缺氧再给氧大鼠心脏微血管内皮细胞粘附分子ICAM－1表达的影响。方法：培养大鼠心脏微血管内皮细胞，建立细胞缺氧再给氧模型，采用酶联免疫吸附试验（ELISA）测定内皮细胞粘附分子ICAM－1的蛋白表达，Northern blot分析测定ICAM－1mRNA的表达。结果：缺氧后内皮细胞ICAM－1的蛋白和mRNA表达明显升高，缺氧再给氧后增高更明显，姜黄素组ICAM－1的蛋白和mRNA的表达较缺氧再给氧组明显下降，呈剂量依赖效应。结论：姜黄素能明显下调缺氧再给氧大鼠心脏微血管内皮细胞ICAM－1的表达，抑制内皮细胞的激活。     

左旋四氢巴马汀对大鼠脑微血管内皮细胞缺氧-复氧时细胞间粘附分子-1的影响

目的：探讨左旋四氢巴马汀（L－THP）对缺氧－复氧时脑微血管内皮细胞细胞间粘附分子－1（ICAM－1）表达的影响。方法：采用流式细胞仪检测脑微血管内皮细胞在缺氧、缺氧－复氧及加入L－THP后ICAM－1的阳性细胞百分率。结果：缺氧12h，脑微血管内皮细胞ICAM－1的表达与常氧时差异无显著意义，缺氧12h再给氧12h,ICAM-1的表达明显增加，加入L－THP后，ICAM－1的阳性细胞百分率明显降低。结论：L－THP可减少缺氧－复氧时脑微血管内皮细胞ICAM－1的表达。     

Reduction of infarct size in the rat heart by LPS preconditioning is associated with expression of angiogenic growth factors and increased capillary density.

Inflammation induces the expression of angiogenic growth factors in tissues, which leads to microvascular growth. Bacterial lipopolysaccharide (LPS) provokes a transient inflammatory response in the heart and induces delayed cardiac resistance to post-ischemic contractile dysfunction. In this study, we examined: 1) the effects of LPS on myocardial expression of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), 2) whether an increase in the density of myocardial microvessels follows the expression of angiogenic growth factors, and 3) the effect of LPS on myocardial resistance to infarction and its relationship with microvascular growth. Rats were treated with LPS (from Salmonella typhimurium, 0.5 mg/kg i.p.). The expression of bFGF and VEGF in the myocardium was examined at 6 and 12 h after LPS treatment by immunofluorescent staining. Myocardial capillary and arteriole densities were determined 3 days after LPS treatment by morphometry, using immunofluorescent staining of von Willebrand factor (a marker protein of endothelial cells) and alpha-smooth muscle actin (a marker protein of smooth muscle cells). To examine cardiac resistance to infarction, hearts were subjected to 40 min of regional ischemia and 2 h of reperfusion by reversible occlusion of left coronary artery at 3 days after LPS treatment. LPS induced cardiac bFGF and VEGF at 6 and 12 h after treatment. The expression of these growth factors was followed by an increase in myocardial capillary density (2032 +/- 78/mm2 vs. 1617 +/- 47/mm2 in saline control, P < 0.05), but not arteriole density, at 3 days. Meanwhile, infarct size was significantly reduced by LPS preconditioning (infarct/left ventricle 12.3 +/- 1.04% vs. 21.7 +/- 1.65% in saline control, 43% reduction, P < 0.05). These results suggest that LPS preconditioning induces cardiac bFGF and VEGF, and an increase in myocardial capillary density. This increased myocardial capillary density is associated with a reduced infarct size after in vivo regional ischemia-reperfusion.     

Dicloxacillin and erythromycin at high concentrations increase ICAM-1 expression by endothelial cells: a possible factor in the pathogenesis of infusion phlebitis

OBJECTIVES: Antimicrobial agents are important risk factors for infusion phlebitis, but the risk varies between different antibiotics. Erythromycin and dicloxacillin are known to induce phlebitis frequently, as well as to exert toxic effects on cultured endothelial cells. The pathogenesis of infusion phlebitis is unclear, but chemical toxicity is thought to lead to inflammation and subsequent thrombosis. In the present study, endothelial cells were exposed to antibiotics at the range of concentrations used for intravenous administration, followed by analysis of pro-inflammatory and pro-coagulant surface molecules. METHODS: Primary human umbilical vein endothelial cells (HUVEC) and the endothelial hybrid cell line EaHy926 were exposed to dicloxacillin, erythromycin, benzylpenicillin and cefuroxime (all at 6250 mg/L) for 60 min, followed by washing. After 5 or 24 h additional incubation, cells were analysed for E-selectin (CD62E), tissue factor (TF) or intercellular adhesion molecule 1 (ICAM-1, CD54) density by flow cytometry. RESULTS: Despite constitutive expression of ICAM-1 (34%) in HUVEC, 6250 mg/L of dicloxacillin or erythromycin significantly increased the number of cells with ICAM-1 expression by 37% and 30%, respectively. In contrast, cefuroxime and benzylpenicillin did not up-regulate ICAM-1 above background levels. A similar pattern was seen with the endothelial cell line EaHy926. The E-selectin and TF density were not affected by the antibiotics examined. CONCLUSIONS: The results of this study support the theory that endothelial cells that are affected by high concentrations of antibiotics may initiate an inflammatory response through expression of ICAM-1. This is a novel finding in the pathogenesis of infusion phlebitis.     

Regulation of intercellular adhesion molecule-1 (ICAM-1) in ischemic and reperfused canine myocardium.

Previous studies in vitro have shown an important role for intercellular adhesion molecule-1 (ICAM-1) in adherence interactions of canine neutrophils with canine jugular vein endothelial cells and in cytotoxicity of canine neutrophils for adult cardiac myocytes. To evaluate the regulation of ICAM-1 in myocardial inflammation and its role in the pathogenesis of myocardial ischemia and reperfusion, a series of in vivo and ex vivo studies were performed in canine animals. Systemic administration of LPS elicited ICAM-1 mRNA in several tissues, including myocardium, which demonstrated increasing ICAM-1 staining on intercalated discs of cardiac myocytes. In ischemia and reperfusion protocols: (a) ICAM-1 mRNA was found in ischemic segments within 1 h of reperfusion and in both ischemic and normally perfused segments by 24 h of reperfusion; (b) expression of ICAM-1 was detected in cardiac myocytes in the ischemic region by 6 h of reperfusion; increased expression was seen thereafter as a function of time; (c) post-ischemic (but not preischemic) cardiac lymph collected at intervals from 1 to 24 h after reperfusion elicited ICAM-1 mRNA, ICAM-1 expression, and ICAM-1-dependent neutrophil adhesion in canine jugular vein endothelial cells and in cardiac myocytes with peak cytokine activity seen by 1 h; (d) extravascular localization of neutrophils was detected in ischemic areas only, and was associated with endothelium bearing high levels of ICAM-1 within 1 h of reperfusion; infiltration increased thereafter in association with increasing levels of ICAM-1 mRNA in myocardial segments and increasing levels of ICAM-1 expression on cardiac myocytes. These findings provide the first direct evidence for inflammatory regulation of ICAM-1 in ischemic and reperfused canine myocardium. They support the hypothesis that ICAM-1 participates in neutrophil-mediated myocardial damage.     

Adherence of neutrophils to canine cardiac myocytes in vitro is dependent on intercellular adhesion molecule-1.

The adhesiveness of isolated canine cardiac myocytes for neutrophils is greatly increased by stimulation with cytokines such as tumor necrosis factor alpha (TNF alpha). Since this adhesion is significantly inhibited by an anti-CD18 MAb, experiments were performed to test the hypothesis that the newly expressed adhesion molecule on the cardiac myocytes was intercellular adhesion molecule-1 (ICAM-1). A newly developed MAb, CL18/6, was found to exhibit the functional and binding characteristics with canine neutrophils and canine jugular vein endothelial cells expected of an antibody recognizing ICAM-1. MAb CL18/6 also bound to isolated cardiac myocytes after stimulation of the myocytes with cytokines, and it blocked by greater than 90% the adhesion of neutrophils to stimulated myocytes. A partial cDNA clone for canine ICAM-1 was isolated, and ICAM-1 mRNA was found to be increased in both endothelial cells and cardiac myocytes after cytokine stimulation. Cytokines that both increased the CL18/6-inhibitable adhesion of neutrophils to myocytes and induced expression of ICAM-1 were IL-1 beta, TNF alpha, and LPS. These results are consistent with the conclusion that canine endothelial cells and cardiac myocytes express ICAM-1 in response to cytokine stimulation, and that ICAM-1 functions as an adhesive molecule for neutrophils on both cell types.     

雌激素相关受体α对脂多糖诱导的内皮细胞凋亡及连接蛋白降解的影响

目的:探讨雌激素相关受体α（ERRα）对脂多糖（LPS）诱导的内皮细胞凋亡及连接蛋白降解的影响。方法:体外培养ERRα敲低的稳转细胞株,并通过LPS处理,将细胞分为四组：正常对照组（Ctr组）;shERRα敲低组（shERRα组）;正常细胞+LPS处理组（LPS组）：六孔板中的细胞用无血清的培养基饥饿培养12 h后,用20 μg/mL LPS处理12 h;shERRα+LPS组：ERRα敲低的细胞按LPS组处理。使用ROS荧光试剂盒检测细胞内ROS的水平;利用TUNEL、AnnexinV-FITC和PI双染检测细胞凋亡情况;细胞荧光检测连接蛋白ZO-1在细胞膜表达情况,Western Blot检测凋亡相关蛋白Bcl-2、Bax、Smac、细胞色素c和连接蛋白ZO-1,Occludin、JAM-A及E-Ca在分子水平的表达情况。结果:与Ctr组及shERRα组相比,LPS组ROS水平增加,细胞凋亡率增加（TUNEL检测为16.44±2.55,流式细胞术检测结果为23.56±2.22）,促凋亡蛋白Bax、Smac及细胞色素c表达量增高,而抗凋亡蛋白Bcl-2和紧密连接蛋白表达量降低,细胞荧光结果显示连接蛋白ZO-1在包膜发生降解,网状结构断裂。而与LPS组相比,在shERRα+LPS组中,抑制ERRα的表达加剧上诉细胞损伤。结论:ERRα能够通过负性调节肺微血管内皮细胞内的细胞凋亡,影响肺微血管内皮的功能,从而调控脓毒症诱导的急性肺损伤。     

脑源性神经营养因子与心脏微血管内皮细胞的管状形成

背景：研究发现脑源性神经营养因子可促进内皮细胞的存活,诱导血管新生,但其促进血管新生的细胞与分子机制尚不清楚。目的：观察脑源性神经营养因子在3D体外管状形成模型中对心脏微血管内皮细胞管状结构形成的影响。方法：分离提取大鼠心脏微血管内皮细胞并培养使之形成细胞小球,将心脏微血管内皮细胞小球接种于Ⅰ型胶原-甲基纤维素的3D环境中培养,然后分别加入50,70,100μg/L的脑源性神经营养因子继续培养,于培养24,48h观察并测量心脏微血管内皮细胞的分枝长度和分枝条数。结果与结论：经分离的大鼠心脏微血管内皮细胞小球在Ⅰ型胶原-甲基纤维素3D环境中培养24h,均可见管状分枝,且脑源性神经营养因子可促进管状分枝生长,以100μg/L的脑源性神经营养因子的促生长效果最明显,分枝也最多。培养至48h,心脏微血管内皮细胞小球的分枝更长。说明脑源性神经营养因子在3D体外管状形成模型中可促进大鼠心脏微血管内皮细胞发芽及管状结构的形成,且具有剂量依赖性。