细胞间黏附分子-1在颅脑损伤中的表达和炎症介导作用

介绍细胞间黏附分子-1（intercellular adhesion molecules-1，ICAM-1）的生化特性和颅脑损伤后介导损伤脑组织炎性反应的作用。另外，许多炎性因子可激活ICAM-1蛋白表达，导致炎症的级联反应。     

实验性家兔动脉粥样硬化形成中细胞间黏附分子-1表达的变化

目的观察细胞间黏附分子（ICAM）-1在动脉粥样硬化（AS）形成中的作用，并试图探讨循环可溶性ICAM-1（sICAM-1）与局部ICAM-1表达的剂量关系。方法雄性新西兰纯种白兔56只，随机分为对照组24只（喂饲普通饲料）和模型组32只（予高脂饮食喂养）；于试验的2、4、6、8周末随机处死每组兔6～8只，取血行血脂及sICAM-1检测，取胸主动脉用免疫组化法检测ICAM-1表达。结果对照组血脂在实验前后无明显变化，而模型组血浆胆固醇、甘油三酯、低密度脂蛋白在2周末时较实验前及对照组即均已明显升高，随实验进展有继续升高趋势。本实验血清中未检测到sICAM-1。免疫组化染色显示，对照组兔血管壁仅内皮细胞ICAM-1呈弱阳性表达，而模型组随高脂饮食时间延长内皮细胞表达增强，且斑块内和中膜平滑肌表达亦逐渐增强、表达范围逐渐增大。结论ICAM-1参与了AS的形成和发展，血清sICAM-1浓度与血管局部ICAM-1间可能为一种高剂量相关关系。     

实验性家兔动脉粥样硬化形成中细胞问黏附分子-1表达的变化

 目的 观察细胞间黏附分子(ICAM)-1在动脉粥样硬化(AS)形成中的作用,并试图探讨循环可溶性ICAM-1(sICAM-1)与局部ICAM-1表达的剂量关系.方法 雄性新西兰纯种白兔56只,随机分为对照组24只(喂饲普通饲料)和模型组32只(予高脂饮食喂养);于试验的2、4、6、8周末随机处死每组兔6～8只,取血行血脂及sICAM-l检测,取胸主动脉用免疫组化法检测ICAM-1表达.结果　对照组血脂在实验前后无明显变化,而模型组血浆胆固醇、甘油三酯、低密度脂蛋白在2周末时较实验前及对照组即均已明显升高,随实验进展有继续升高趋势.本实验血清中未检测到sICAM-1.免疫组化染色显示,对照组兔血管壁仅内皮细胞ICAM-1呈弱阳性表达,而模型组随高脂饮食时间延长内皮细胞表达增强,且斑块内和中膜平滑肌表达亦逐渐增强、表达范围逐渐增大.结论 ICAM-1参与了AS的形成和发展,血清sICAM-1浓度与血管局部ICAM一1间可能为一种高剂量相关关系.     

α-硫辛酸对高糖作用下血管内皮细胞ICAM-1表达的影响

目的 观察α-硫辛酸对高糖作用下血管内皮细胞的细胞间黏附分子-1(ICAM-1)表达的影响.方法 实验分3组:正常对照组(NG,5.5mmol/L葡萄糖);高糖组(HG,30mmol/L葡萄糖);药物干预组[30mmol/L葡萄糖+不同浓度α-硫辛酸(50、100、200μmol/L)].脐静脉内皮细胞株在不同条件下孵育48h,采用黄嘌呤氧化酶法测定各组内皮细胞SOD活性,以硫代巴比妥酸为底物检测MDA含量,ELISA法测定各组内皮细胞ICAM-1蛋白浓度,RT-PCR法检测内皮细胞ICAM-1 mRNA表达水平.结果 与NG组相比,HG组内皮细胞SOD活性明显降低(P<0.01),MDA含量、ICAM-1蛋白及mRNA表达水平明显上升(P<0.01);与HG组相比,药物干预组内皮细胞SOD活性明显升高(P<0.01),MDA含量、ICAM-1蛋白及mRNA表达水平明显降低(P<0.05).结论 α-硫辛酸可改善高糖作用下血管内皮细胞的氧化应激,抑制内皮细胞ICAM-1的表达水平.     

大鼠缺血再灌注后小动脉ICAM-1与NOS表达关系的实验研究

目的：观察大鼠缺血再灌注后细胞间黏附分子（ICAM-1）和一氧化氮合酶（NOS）表达及其相互关系。方法：将大鼠分为对照组、缺血45min组和缺血再灌注1、3、6、12、24h组,建立失血性休克模型并给予治疗,用光镜和电镜观察小动脉形态学改变,并用免疫组化法检测ICAM-1、NOS在小动脉内皮和平滑肌细胞表达的阳性细胞数。结果：（1）缺血45min后NOS表达量首先增加,再灌后1h达到高峰,3h表达减弱。与对照组比较,差异均有统计学意义（P〈0．05）;（2）缺血再灌后1h组ICAM-1表达量增加,再灌后3h达到高峰,并持续至24h,与对照组比较,差异均有统计学意义（P〈0．05）。结论：ICAM-1介导了小动脉缺血再灌注损伤,NOS对小动脉缺血再灌注损伤具有保护作用,NOS对ICAM-1有抑制作用。     

细胞间黏附分子-1在肝癌组织中表达及临床意义

目的探讨细胞间黏附分子-1(ICAM-1)在肝癌组织中的表达及临床意义。方法选取自2005年8月至2011年8月收治的68例原发性肝癌术后患者的肝细胞肝癌组织及癌旁组织标本为研究对象。采用免疫组织化学染色方检测肝癌患者的癌组织及癌旁组织中ICAM-1的表达。结合患者病理指标、病理分型分析ICAM-1分子表达的临床意义。结果免疫组化结果显示,肝癌组织ICAM-1表达显著高于癌旁组织,差异有统计学意义(P<0.05)。ICAM-1表达水平与分化程度、临床分期、肿瘤个数、甲胎蛋白、是否远处转移相关(P<0.05);与患者年龄、性别、乙肝表面抗原、肿瘤直径无关(P>0.05)。CAM-1低表达中位总体生存期为45个月,显著高于高表达患者的32个月,差异有统计学意义(P<0.05)。结论 ICAM-1在肝癌组织中呈高表达,且其表达水平与肝癌的临床分期、转移能力和临床预后密切相关,是肝癌潜在的预后判断指标。     

细胞间黏附分子-1和CD44s在食管癌组织中的表达及临床意义

目的：研究细胞间黏附分子-1(ICAM-1)及CD44s mRNA在食管癌组织及癌旁正常组织的表达情况及其临床意义。方法：采用RT-PCR的方法检测新鲜收集的食管癌组织及癌旁正常组织中ICAM-1及CD44s的表达。结果：食管癌组织中ICAM-1和CD44s的表达水平明显高于癌旁组织。ICAM-1和CD44s的高水平表达与食管癌TNM分期及淋巴结转移之间有明显的关系，而与分化程度无关。结论：食管组织在恶变过程中ICAM-1和CD44s mRNA表达水平明显增高，提示二者在食管癌的发生、发展中起重要作用。     

低强度氦氖激光对小鼠骨髓基质细胞黏附分子表达的影响

目的研究低强度氦氖(He-Ne)激光照射对小鼠骨髓基质细胞(BMSCs)表面细胞间黏附分子-1(ICAM-1)、血管细胞间黏附分-子1(VCAM-1)表达的影响。方法 30只小鼠随机分为3组:对照组(A组),2.75J/cm2激光组(B组),4.20J/cm2激光组(C组),每组10只。激光组建立低强度He-Ne激光体外照射小鼠模型,对照组不作任何处理。分离小鼠BMSCs,并进行原代、传代培养。采用倒置相差显微镜、光镜及透射电镜对BMSCs形态进行观察;采用流式细胞术检测低强度He-Ne激光照射对小鼠BMSCs表面VCAM-1和ICAM-1蛋白表达的影响。结果倒置显微镜和光镜观察到BMSCs呈贴壁生长,细胞呈梭形或不规则,有突起;电镜观察到细胞内粗面内质网、分泌小泡、线粒体等细胞器丰富。激光组BMSCs增殖加快,达到80%细胞融合时间短。2.75J/cm2激光组和4.20J/cm2激光组均能增加小鼠BMSCs表面VCAM-1和ICAM-1蛋白的表达。其中,4.20J/cm2激光组较2.75J/cm2激光组作用更明显(P<0.01)。结论低强度He-Ne激光能促进小鼠BMSCs表面VCAM-1和ICAM-1蛋白的表达。     

缺血预适应对大鼠肢体缺血再灌注后肺损伤时黏附分子表达的影响

目的 探讨缺血预适应对大鼠肢体缺血再灌注致肺损伤时黏附分子ICAM-1表达的影响.方法 雄性Wistar大鼠30只,随机分为对照组、缺血再灌注(IR)组和缺血预处理(IPC+IR)组,每组10只.采用流式细胞技术、反转录聚合酶链反应(RT-PCR)和Western blotting等观察肢体缺血再灌注后肺损伤发生过程中,血液中CD18阳性的多形核白细胞(PMN)百分率,以及肺组织ICAM-1在mRNA和蛋白水平的变化;测定肺组织湿/干重值(W/D);检测肺组织中髓过氧化物酶(MPO)含量;观察肺组织形态学的改变及缺血预适应对上述指标的影响.结果 大鼠肢体缺血再灌注后W/D及CD18阳性细胞数均明显增加,肺组织MPO活性增加,ICAM-1表达明显升高,预适应可以明显抑制这些变化,从而对肺起一定保护作用.结论 缺血预适应对肢体缺血再灌注后肺损伤的保护作用与下调黏附分子表达有关.     

血清可溶性细胞间黏附分子-1与甲状腺相关眼病的相关性研究

 目的探讨血清可溶性细胞间黏附分子-1(sICAM-1)与甲状腺相关眼病(Thyroid associated ophthalmopathy,TAO)及其活动性的关系.方法运用ELISA方法,检测伴或不伴TAO的Graves病患者及正常对照组患者的血清sICAM-1水平;活动性和非活动性TAO及正常对照组的血清sICAM-1水平.结果发现伴或不伴TAO的Graves病患者的sICAM-1水平均高于正常对照组(426.80±77.19 ng/ml).而且,伴有TAO的Graves病患者sICAM-1水平高于不伴TAO的Graves病患者,两者差异有显著性(P＜0.01).活动性TAO组sICAM-1水平(1127.10±190.01 ng/ml)显著高于非活动性TAO组(638.30±86.44ng/ml),两者差异有显著性(P＜0.01),且均高于正常对照组.结论TAO患者眼眶组织免疫反应产生的ICAM-1是血清sICAM-1的重要来源.血清sICAM-1水平与TAO眼眶免疫病变的活动程度密切关联,可作为监测眼眶免疫病变活动的指标.     

Post-mortem markers of sepsis: an immunohistochemical study using VLA-4 (CD49d/CD29) and ICAM-1 (CD54) for the detection of sepsis-induced lung injury

The up-regulation of different adhesion molecules such as VLA-4 (CD49d/CD29) and ICAM-1 (CD54) on the pulmonary endothelium and leukocytes, is a key event in sepsis-induced lung injury leading to inflammatory tissue alterations. The value of VLA-4 and ICAM-1 as micromorphological post-mortem markers for the detection of sepsis-induced lung injury, was evaluated in a semiquantitative immunohistochemical study. VLA-4 was strongly expressed on intravascular, interstitial and intra-alveolar leukocytes in sepsis-associated fatalities, whereas in non-septic fatalities an irregular weak immunoreactivity was observed on interstitial leukocytes and no positive immunohistochemical expression was detected on intravascular or intra-alveolar leukocytes. ICAM-1 was strongly expressed on endothelial cells of the pulmonary microvasculature and on pulmonary macrophages and lymphocytes in sepsis-associated fatalities. In contrast, an infrequent weak immunohistochemical reaction for ICAM-1 was found on pulmonary endothelium and on perivascular leukocytes in non-septic fatalities. Based on the results of the present preliminary study, VLA-4 and ICAM-1 can be considered as useful immunohistochemical post-mortem markers of sepsis. Received: 28 February 2000 / Accepted: 22 May 2000     

Magnetic resonance imaging of radiation-induced brain injury using targeted microparticles of iron oxide

Background Radiation-induced brain injury (RBI) is the most serious complication of primary and metastatic brain and neck malignant tumors following radiation therapy. However, at present, RBI is difficult to diagnose in the early period. Recently, studies have demonstrated that the early stage of RBI is characterized by an inflammatory reaction, and that intercellular adhesion molecule-1 (ICAM-1) is significantly up-regulated in the irradiated brain tissues. Purpose To provide an early diagnosis of RBI using molecular magnetic resonance imaging (MRI) with microparticles of iron oxide (MPIO) targeted to ICAM-1 in the vascular endothelium of brains. Material and Methods A monoclonal antibody against ICAM-1 was conjugated to MPIO to form the targeted MRI contrast agent ICAM-MPIO. The adhesion of ICAM-MPIO to endothelial cells was quantified by optical imaging and MRI. Sprague-Dawley rats were irradiated to establish an animal model of the early period of RBI. ICAM-MPIO and free-MPIO were injected via tail vein, respectively. T(2) signal intensity and T(2) values of the irradiated brains and normal brains were subsequently evaluated by MRI. Results In vitro, the adhesion of ICAM-MPIO to the activated endothelial cells was 5 ± 0.5-fold greater than to the non-stimulated cells, which could be detected by optical imaging and MRI (R(2) = 1.0, P < 0.01). In vivo, ICAM-MPIO caused a marked negative MRI contrast effect in irradiated brains. As compared with brains without irradiation, the specific contrast effect increased more than seven-fold after administration of ICAM-MPIO (F = 751.495, P < 0.05). Conclusion MPIO coated with monoclonal antibody of ICAM-1 could be used for detecting the early period of RBI by optical imaging and MRI.     

蒺藜总皂苷对大鼠缺血再灌注损伤心肌ICAM-1表达及中性粒细胞浸润的影响

目的观察蒺藜总皂苷对缺血再灌注损伤心肌炎症因子ICAM-1表达及中性粒细胞浸润的影响。方法可逆性冠脉左前降支结扎缺血45min再灌注2h复制MI／R模型,SD雄性大鼠32只,随机分成假手术组、模型组、GSTT大剂量组、GSTT小剂量组,每组8只。HE染色观察心肌中性粒细胞浸润,测定心肌MPO活性,免疫组化测定心肌组织ICAM-1的表达。结果HE染色镜下观察模型组心肌中性粒细胞浸润严重,而GSTT大、小剂量组则较轻;GSTT大、小剂量组MPO活性显著降低（P〈0．01或0．05）;缺血45min再灌注2h心肌组织ICAM-1无明显表达。结论GSTT对MI／R炎症损害有保护作用,可降低IVIPO活性,减轻中性粒细胞浸润。     

Differences in ICAM-1 and TNF-alpha expression between large single fraction and fractionated irradiation in mouse brain

PURPOSE: To elucidate the brain molecular response to irradiation. The expression of the intercellular adhesion molecule (ICAM-1) and tumour necrosis factor-alpha (TNF-alpha) in the mouse brain was compared after single-dose and fractionated whole-brain irradiation. MATERIALS AND METHODS: Mice received a single dose of 2, 10 or 20 Gy or a fractionated dose (2 Gy day(-1)) of 10, 20 or 40 Gy. ICAM-1, and TNF-alpha mRNA expression were quantified by the highly sensitive real-time polymerase chain reaction technique. Expression of ICAM-1 protein was quantified by dual-labelled monoclonal antibody assay. RESULTS: After a 20-Gy single dose, there was an increase in ICAM-1 and TNF-alpha mRNA levels (14- and 11-fold, respectively) as well as a significant increase in the level of ICAM-1 protein (p=0.0243). The expression of ICAM-1 and TNF-alpha mRNA increased at the end of the 40-Gy fractionated regimen (3.55- and 2.30-fold, respectively). CONCLUSIONS: The molecular response of the brain to single-dose irradiation was rapid, while its response to fractionated irradiation was slow. This finding is consistent with clinical observations and could be of use when designing strategies to mitigate radiation sequelae.     

Molecular MR Imaging for Visualizing ICAM-1 Expression in the Inflamed Synovium of Collagen-Induced Arthritic Mice

Objective

To determine the utility of intercellular adhesion molecule (ICAM)-1 antibody-conjugated gadolinium diethylenetriaminepentaacetic acid (Gd-DTPA-anti-ICAM-1) as a targeted contrast agent for the molecular magnetic resonance imaging (MRI) in collagen-induced arthritis (CIA).

Materials and Methods

Three groups of mice were used: non-arthritic normal, CIA mice in both the early inflammatory and chronic destructive phases. The MR images of knee joints were obtained before and after injection of Gd-DTPA-anti-ICAM-1, Gd-DTPA, and Gd-DTPA-Immunoglobulin G (Ig G) and were analyzed quantitatively. The patterns of enhancement on the MR images were compared with the histological and immunohistochemical ICAM-1 staining.

Results

The images obtained after injection of Gd-DTPA-anti-ICAM-1 displayed gradually increasing signal enhancement from the moment following injection (mean ± standard deviation [SD]: 424.3 ± 35.2, n = 3) to 24 hours (532 ± 11.3), rather than on pre-enhanced images (293 ± 37.6) in the early inflammatory phase of CIA mice. However, signal enhancement by Gd-DTPA and Gd-DTPA-IgG disappeared after 80 minutes and 24 hours, respectively. In addition, no significant enhancement was seen in the chronic destructive phase of CIA mice, even though they also showed inflammatory changes on T2-weighted MR images. ICAM-1 expression was demonstrated in the endothelium and proliferating synovium of the early inflammatory phase of CIA mice, but not in the chronic destructive phase.

Conclusion

Molecular MRI with Gd-DTPA-anti-ICAM-1 displays specific images targeted to ICAM-1 that is expressed in the inflamed synovium of CIA. This novel tool may be useful for the early diagnosis and differentiation of the various stages of rheumatoid arthritis.     

小鼠放射性肺损伤中细胞间黏附因子-1的表达

目的利用小鼠放射性肺损伤模型，了解细胞间黏附因子-1（ICAM-1）在放射性肺损伤不同阶段中的表达及其意义。方法成年雌性C57BL／6小鼠90只，按随机表分作2组：①对照组（36只）：不做任何处理；②照射组（54只）：全肺单次照射12Gy。于照射后1、24和72h及1、2、4、8、16和24周取照射组及相同鼠龄对照组小鼠肺组织，分别用HE和Masson染色在光镜下观察组织病理改变和胶原纤维沉积；免疫组织化学S-P法观察ICAM-1蛋白和白细胞共同抗原（CD45）蛋白表达，用碱水解法测定羟脯氨酸含量和实时定量RT—PCR法检测ICAM-1 mRNA相对含量。结果光镜下，照射组小鼠肺组织较对照组组织学改变和胶原纤维沉积明显，免疫组织化学结果显示，对照组小鼠肺组织中ICAM-1和炎性反应细胞的阳性细胞数均相对较低，照射组的阳性细胞数较之明显增高（P〈0．01）；羟脯氨酸含量的测定结果显示，照射组小鼠肺组织中的羟脯氨酸含量较对照组明显增高（P〈0．05）；实时定量RT-PCR的结果显示，照射组的ICAM-1 mRNA的表达水平较对照组均明显升高（P〈0．01）。结论在放射性肺损伤的过程中，ICAM-1是一重要的细胞因子，不仅介导早期炎性反应细胞的黏附和浸润，还参与后期放射性肺纤维化的形成。     

两种浓度七氟烷联合舒芬太尼后处理对创伤性失血性休克炎性反应的影响

目的 评估舒芬太尼与七氟烷联合后处理对创伤性失血性休克患者炎性反应的影响.方法 成年创伤性失血性休克120例随机分成6组：S组、C1组、C2组在输血前30 min经静脉给予0.1 μg/kg舒芬太尼.S1、C1组与S2、C2组输血前30 min分别吸入0.5 MAC（1MAC=1.7%）及1 0 MAC七氟烷10 min.C组为对照组未行后处理.在给予舒芬太尼前（T0）、给予舒芬太尼后30 min（T1）、七氟烷处理后2 h（T2）和24 h（T3）采血测定TNF-α、IL-6、IL-8、ICAM-1浓度.结果 T2时,S、S2、C1、C2组TNF-α、IL-6、IL-8、ICAM-1浓度较C组差异有统计学意义,并且C2组与其他试验组间差异亦有统计学意义（P＜0.05）.T3时,S2、C1、C2组IL-6浓度较T0和C组差异有统计学意义（P＜0.05）; C1、C2组IL-8浓度较C组差异有统计学意义（P＜0.05）.结论 七氟烷舒芬太尼联合后处理通过降低炎性因子的水平,减轻创伤性失血性休克炎性反应,从而发挥脏器保护的作用.     

siRNA干扰明胶酶对人肾小管细胞ICAM-1表达的影响

目的 利用小干扰RNA(siRNA)抑制人肾小管上皮细胞(HKC)的基质金属蛋白酶-2(MMP-2)和MMP-9,即明胶酶A和B的表达,观察其对胞间黏附分子1(ICAM-1)表达的影响.方法 培养人肾小管上皮细胞,以100nmol/L佛波醇酯(PMA)刺激18h,脂质体转染抑制MMP-2或MMP-9表达的siRNA或无关siRNA.提取细胞总RNA或胞浆蛋白,利用Real-Time PCR、Western blot或免疫荧光结合激光共聚焦显微镜技术检测MMP-2、MMP-9或ICAM-1的表达情况.结果 特异性的siRNA能有效抑制HKC中MMP-2、MMP-9的mRNA和蛋白表达;免疫荧光结果显示经PMA刺激后HKC的ICAM-1表达上调,同时MMP-9-siRNA转染组人肾小管细胞的ICAM-1蛋白表达水比其他实验组高(P＜0.05).结论 抑制肾小管上皮细胞MMP-9的表达可使ICAM-1表达上调,提示除了细胞外基质降解途径外,MMP-9还可通过炎症途径影响肾脏纤维化进程.