脊髓缺血-再灌注损伤细胞间粘附分子-1及白细胞介素-1β的表达

目的:探讨脊髓缺血-再灌注损伤细胞间粘附分子-1(ICAM-1)及白细胞介素-1β(IL-1)的表达对血脊髓屏障损害的分子机制。方法:将77只同龄Wistar大鼠,随机分为正常对照组、单纯缺血组和缺血再灌组。手术方法采用Zivin法复制模型。应用逆转录-聚合酶链反应、地高辛标记cDNA探针技术、免疫组化及免疫荧光激光共聚焦扫描显微镜技术检测脊髓再灌注损伤血管内皮ICAM-1mRNA和IL-1βmRNA表达量。结果:正常组和单纯缺血组未引起细胞因子和粘附分子表达量的增加。而缺血再灌注后缺血区细胞因子、粘附分子的表达及多形核白细胞(PMN)的浸润先后发生了改变。再灌注2h,IL-1βmRNA的表达首先升高,约为对照组的2倍。再灌注6h达到高峰,并持续到12h。ICAM-1mRNA表达量于再灌注4h明显升高,再灌注12h其在单位微血管面积上的荧光强度约比单纯缺血组增加了1/2。结论:再灌注损伤后脊髓微血管内皮ICAM-1及其调节因子IL-1β的表达量增加是导致血脊髓屏障损害的重要分子基础。     

Protective effects of monoclonal antibody to intercellular adhesion molecule-1 in venous ischemia-reperfusion injury: experimental study in rats

Flaps with venous occlusion have a decreased survival rate compared with arterial occlusion. It seems that several factors are involved in the etiology of total venous occlusion, including free radicals, edema, thrombosis, and reperfusion injury. In the present study, the authors evaluated the blockage of polymorphonuclear leukocyte endothelial adhesion by using a monoclonal antibody to the intercellular adhesion molecule 1 (ICAM-1) ligand to prevent venous ischemia-reperfusion injury in rat epigastric island flaps. A skin flap (3 x 4 cm) supplied by the superficial epigastric artery and vein was harvested unilaterally in 40 male Wistar rats. Total venous occlusion of the skin flap was achieved. Arterial inflow was left intact. Rats were randomly divided into four groups (n = 10). In Group 1; rats were intravenously pretreated with 0.5 ml of 0.9 percent normal saline 15 min before applying a venous clamp, and the flaps were subjected to 6 hr of venous ischemia. In Group 2; rats were intravenously pretreated with 0.05 mg of monoclonal antibody to the intercellular adhesion molecule 1 (0.20 mg/kg) in 0.5 ml of 0.9 percent normal saline 15 min before applying the venous clamp, and the flaps were subjected to venous ischemia as in Group 1. In Group 3; rats were pretreated as in Group 1, and the flaps were subjected to 8 hr of venous ischemia. In Group 4; rats were pretreated as in Group 2, and the flaps were subjected to 8 hr of venous ischemia. The flaps were assessed histologically and by measuring viable and non-viable areas on postoperative day 7. Flap measurements revealed that blocking the action of ICAM-1 IN VIVO by administering monoclonal antibody significantly attenuated ischemic injury after 6 or 8 hr of venous occlusion.     

Clinical effects of inhibiting leukocyte adhesion with monoclonal antibody to intercellular adhesion molecule-1 (enlimomab) in the treatment of partial-thickness burn injury

BACKGROUND: A randomized, prospective, multicenter, double-blind, placebo-controlled, phase II clinical trial was performed to determine whether inhibition of leukocyte adherence by administration of monoclonal antibody directed against intercellular adhesion molecule-1 would improve burn wound healing. METHODS: One hundred ten patients with burn injury ranging from 10% to 30% total body surface area were enrolled. Fifty-six patients received placebo (saline) and 54 patients received murine monoclonal antibody to the human intercellular adhesion molecule-1 (enlimomab). Treatment was initiated within 6 hours of injury. Patients had three distinct partial-thickness wound sites assessed. Laser Doppler flowmetry was used to stratify wounds on the day of injury. Wounds were assessed for healing status on day 21 postburn and categorized as healed, nonhealed, or grafted. RESULTS: Patients treated with enlimomab had a significantly increased percentage of wounds that healed spontaneously in less than 21 days overall and when stratified by burn wound laser Doppler blood flow readings for those wounds at greatest risk for nonhealing. CONCLUSION: These results support the concept that leukocyte adherence is involved in the pathogenesis of burn wound necrosis and suggest a therapeutic mechanism for modulating the inflammatory response after the burn injury that may improve wound healing.     

Attenuation of postlaminectomy epidural fibrosis with monoclonal antibodies against intercellular adhesion molecule-1 and CD-18.

BACKGROUND CONTEXT: Data from studies in other diseases state implicate cellular adhesion molecules as mediators of fibrosis and scarring. We sought to explore and assess the effect of using monoclonal antibodies against intercellular adhesion molecule-1 (ICAM-1) and its ligand CD-18 to decrease epidural fibrosis in an animal spinal surgery model. PURPOSE: We hypothesize that use of antiadhesion molecules (anti-ICAM-1 and anti-CD-18) decreases epidural fibrosis in rats after spinal surgery compared with nontreated group and monoclonal anti human immunoglobulin (Ig)G group. STUDY DESIGN: Experimental animal spine surgery (laminectomy) protocol with application of antiadhesion molecules (anti-ICAM-1 and anti-CD-18 group as a specific monoclonal antibody) to surgical site in test group compared with monoclonal antihuman IgG group (as a nonspecific monoclonal antibody) and nontreated group. METHODS: Thirty Sprague Dawley male or female rats weighing 175 to 250 g were used randomly for three groups (nontreated, anti-ICAM-1 and anti-CD-18, monoclonal antihuman IgG). Laminectomy was performed at level L4 in all animal groups. After injection of materials (except nontreated group), the surgical sites were closed in layers. Three weeks later, all rats were killed. Twenty-seven rats were available for histological analysis. The histological sections were evaluated for fibroblast numbers of fibrous tissue within the laminectomy side, adhesion degree between dura mater and fibrous tissue, and new bone formation in the laminectomy region. RESULTS: Comparing the fibroblast numbers in fibrous tissue within groups, the number of fibroblasts were significantly less in anti-ICAM-1 and anti-CD-18 group than nontreated group (p=.037). The number of fibroblasts of monoclonal anti human IgG group was not significantly different from anti-ICAM-1 and anti-CD-18 (p=.608) and the nontreated group (p=.508). In the anti-ICAM-1 and anti-CD-18 applied group, adhesion degree was found significantly less than monoclonal antihuman IgG (p=.036) and nontreated group (p=.036) statistically. There were no significant difference between the monoclonal antihuman IgG group and the nontreated group about adhesion degree (p=.645). CONCLUSIONS: Therapy that targets ICAM-1 could be valuable in the management of epidural fibrosis. Blocking the function of ICAM-1 may provide cellular protection against epidural fibrosis and also it may serve as an important component in this period, acting to promote leukocyte migration across epidural area after laminectomy.     

Hepatic ischemia/reperfusion injury in P-selectin and intercellular adhesion molecule-1 double-mutant mice

Neutrophil adhesion and recruitment represents one of the early cellular events that occur during hepatic ischemia/reperfusion (IR) injury and plays a critical role in determining the extent of tissue damage. The adhesion molecules, such as selectins and intercellular adhesion molecules (ICAM), are important in mediating neutrophil-endothelial cell interactions and neutrophil emigration. The goal of this study was to evaluate the role of P-selectin and ICAM-1 in hepatic IR injury. Male wild-type and P-selectin/ICAM-1-deficient (P/I null) mice underwent 90 minutes of partial hepatic ischemia followed by reperfusion at various time points (0, 1.5, 3, and 6 hours). Reperfusion caused a time-dependent hepatocellular injury in both wild-type and P/I null mice as judged by plasma alanine aminotransferase (ALT) levels and liver histopathology examination. Although ALT levels were slightly lower in the P/I null mice compared with the wild-type mice the differences were not statistically significant. Neutrophil infiltration to the ischemic liver was observed in both mouse groups after 6 hours of reperfusion; however, the infiltration to the midzonal region of the ischemic liver was more pronounced in the wild-type group. This study suggests that hepatocellular injury induced after hepatic IR was independent of P-selectin and ICAM-1 in this model of acute inflammatory tissue injury.     

Evaluation of rapid ischemic preconditioning in a rabbit model of spinal cord ischemia

BACKGROUND: Rapid ischemic preconditioning (IPC) has been shown to reduce cellular injury after subsequent cardiac and cerebral ischemia. However, the data on rapid IPC of the spinal cord is limited. The authors investigated whether pretreatment with sublethal ischemia of spinal cord can attenuate neuronal injury after spinal cord ischemia in rabbits. METHODS: Forty-seven male New Zealand white rabbits were randomly assigned to one of three groups (n = 15 or 16 each). In the IPC(-) group, the infrarenal aorta was occluded for 17 min to produce spinal cord ischemia. In the IPC(+) group, 5 min of aortic occlusion was performed 30 min before 17 min of spinal cord ischemia. In the sham group, the aorta was not occluded. Hind limb motor function was assessed at 3 h, 24 h, 4 days, and 7 days after reperfusion using Tarlov scoring (0 = paraplegia; 4 = normal). Animals were killed for histopathologic evaluation at 24 h or 7 days after reperfusion. The number of normal neurons in the anterior spinal cord (L4-L6) was counted. RESULTS: Neurologic scores were significantly higher in the IPC(+) group than the IPC(-) group at 3 and 24 h after reperfusion (P < 0.05). However, neurologic scores in the IPC(+) group gradually decreased and became similar to those in the IPC(-) group at 4 and 7 days after reperfusion. At 24 h after reperfusion, the numbers of normal neurons were significantly higher in the IPC (+) group than in the IPC(-) group (P < 0.05) and were similar between the IPC(+) and sham groups. At 7 days after reperfusion, there was no difference in the number of normal neurons between the IPC(+) and IPC(-) groups. CONCLUSION: The results indicate that rapid IPC protects the spinal cord against neuronal damage 24 h but not 7 days after reperfusion in a rabbit model of spinal cord ischemia, suggesting that the efficacy of rapid IPC may be transient.     

The effect of thalidomide on spinal cord ischemia/reperfusion injury in a rabbit model

STUDY DESIGN: Randomized study. OBJECTIVES: To evaluate the effects of thalidomide on spinal cord ischemia/reperfusion injury via reduced TNF-alpha production. SETTING: Animal experimental laboratory, Clinical Research Institute of Seoul National University Hospital, Seoul, Korea. METHODS: Spinal cord ischemia was induced in rabbits by occluding the infrarenal aorta. Rabbits in group N did not undergo ischemic insult, but rabbits in groups C (the untreated group), THA, and THB underwent ischemic insult for 15 min. The THA and THB groups received thalidomide (20 mg/kg) intraperitoneally (i.p.) before ischemia, but only the THB group received thalidomide (i.p., 20 mg/kg) after 24 and 48 h of reperfusion. After evaluating neurologic functions at 1.5 h, 3, and 5 days of reperfusion, rabbits were killed for histopathologic examination and Western blot analysis of TNF-alpha. RESULTS: The THA and THB groups showed significantly less neurologic dysfunction than the C group at 1.5 h, 3, and 5 days of reperfusion. The number of normal spinal motor neurons in ventral gray matter was higher in THA and THB than in C, but no difference was observed between THA and THB. Western blot analysis showed a significantly higher level of TNF-alpha in C than in THA and THB at 1.5 h of reperfusion, but no difference was observed between C, THA, or THB at 3 or 5 days of reperfusion. CONCLUSION: Thalidomide treatment before ischemic insult reduces early phase ischemia/reperfusion injury of the spinal cord in rabbits.     

Increased expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 in rat cardiac allografts

This study was designed to investigate the role of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) during chronic cardiac allograft rejection. Wistar rats were used as donors, and SD rats as recipients heterotopic cardiac transplants. Recipients pretreated with inoculation of donor splenocytes (SPC) followed by cyclophosphamide (CP) were divided into 4 groups: (A) untreated group (n = 18) without immunosuppression; (B) SPC plus CP-treated group (n = 18) that were euthanized at 15-120 days posttransplantation; (C) CsA-treated group (n = 18) euthanized at 2-3 months posttransplantation; and (D) tolerance group (n = 18) treated with SPC plus CP and monitored for at least 1 year posttransplantation. Cardiac allografts were harvested at various times for immunohistochemical studies performed to evaluate the expression of ICAM-1 and VCAM-1. Pretreatment of animals with SPC and CP induced long-term cardiac allograft survival. Immunohistochemical staining demonstrated a low level of ICAM-1 and VCAM-1 expression in cardiac allograft muscle and coronary arteries among Groups B and D. In contrast, the expressions of ICAM-1 and VCAM-1 in cardiac allografts of Groups A and C were significantly higher than those in Groups B and D. Our results suggested that the expression of ICAM-1 and VCAM-1 plays an important role during the development of chronic cardiac allograft rejection.     

Experimental spinal epidural abscess: a pathophysiological model in the rabbit

To define the pathophysiology of spinal cord dysfunction associated with spinal epidural abscess formation, we developed an experimental model. Spinal epidural abscesses were produced in rabbits by injecting Staphylococcus aureus into the posterior thoracolumbar epidural space under direct vision. Progressive neurological deficits were detected in 18 of 20 animals; severe paraparesis or paraplegia occurred in 75%, and sphincter dysfunction occurred in 55%. Clinical data, including the results of plain spine roentgenography, myelography, and biochemical and bacteriological examination of the cerebrospinal fluid, were recorded. Epidural abscesses with varying degrees of spinal cord compression were confirmed pathologically in 95% of the experimental group. Spinal cord white matter changes included vacuolization, loss of myelin, and axonal swelling. The gray matter of the spinal cords was relatively preserved. There was no microscopic evidence of thrombosis or vasculitis in the major blood vessels supplying the spinal cords. Histopathological changes detected in the spinal cords were more consistent with direct compression of neural tissue than with infarction. The progressive clinical course and the histopathological changes in the spinal cord after compression by abscess closely resembled those of experimental compression of the spinal cord by epidural neoplasm.     

Clinical significance of intercellular adhesion molecule-1 in ulcerative colitis

This study was conducted to assess the clinical significance of intercellular adhesion molecule-1 (ICAM-1) in ulcerative colitis (UC). The subjects were 53 cases of UC and 43 control cases. ICAM-1 was expressed to a greater degree in the UC specimens. Serum ICAM-1 concentrations in the UC group showed values that were lower than those in the control group (P = 0.0081). Serum ICAM-1 concentrations were found to vary according to degree of clinical severity, activity, and affected range. Considering the cases of surgery, there was only one case in which a lowering of early postoperative values was found. In all cases in which therapeutic drugs including steroids were not administered, late postoperative values were significantly increased. ICAM-1 dynamics in UC were frequently seen in tissue and exhibited a significantly low value in blood serum; however, the influence of clinical severity, activity, diseased area, measuring time, and therapeutic drugs need further study.     

P选择素、细胞间黏附分子-1在大鼠胰腺移植缺血再灌注损伤中的作用

目的观察P选择素（Ps）和细胞间黏附分子-1（ICAM-1）在大鼠胰腺移植缺血再灌注损伤中的作用及Ps单克隆抗体的治疗作用。方法75只SD大鼠随机分为假手术组、移植组和治疗组。移植组和治疗组均进行全胰十二指肠移植术，但治疗组于再灌注前5rain静脉注射Ps单克隆抗体。3组分别于腹主动脉开放后1（n=5）、3（n=5）、6（n=5）h取血测定血清淀粉酶水平，并切取胰腺标本进行组织病理学观察及Ps、ICAM．1免疫组织化学染色。结果移植组胰腺组织损伤随再灌注时间的延长而加重，血淀粉酶升高，与中性粒细胞浸润直接相关；而治疗组胰腺组织损伤不明显，血淀粉酶减低。移植组各时段Ps、ICAM-1均有阳性表达，且Ps再灌注1h为表达高峰，ICAM．1随再灌注时间延长表达逐渐增加，假手术组、治疗组Ps、ICAM-1不表达。结论Ps及ICAM-1按一定时间顺序参与胰腺移植缺血再灌注损伤的病理过程；Ps可能是胰腺移植缺血再灌注损伤的起始因素；抗Ps单克隆抗体对移植胰腺缺血再灌注损伤有保护作用。     

缺血再灌注对大鼠肺细胞间粘附分子-1表达的影响

目的 观察缺血再灌注（Ｉ／Ｒ）对肺组织细胞间粘附分子－１（ＩＣＡＭ－１）表达的影响，分析ＩＣＡＭ－１表达与肺内中性粒细胞浸润的关系。方法 Ｗｉｓｔａｒ大鼠单肺原位热缺血再灌注损伤模型分７组，左肺缺血９０ｍｉｎ，再灌注时间分别为０、１、２、４、８、１６、２４ｈ。逆转录－聚合酶链式反应（ＲＴ－ＰＣＲ）法及Ｗｅｓｔｅｒｎ ｂｌｏｔ法检测肺组织ＩＣＡＭ－１ｍＲＮＡ及蛋白表达，测定各组肺组织髓过氧化物酶活性     

Mesangial expression of intercellular adhesion molecule-1 in primary glomerulosclerosis.

Frozen sections of renal biopsy specimens from eight patients with primary focal segmental glomerulosclerosis (FSGS) and 10 patients with membranous nephropathy (MN) were stained in immuno-peroxidase with the intercellular adhesion molecule-1 (ICAM-1) monoclonal antibody (MoAb), CL203.4. ICAM-1 was expressed by mesangial cells in six patients with FSGS. On the other hand, ICAM-1 was not detected in mesangial cells in patients with MN or in the non-affected portion of tumoral kidneys used to control normal renal expression of ICAM-1. De novo mesangial expression of ICAM-1 in FSGS suggests that sclerosis results from an inflammatory process, possibly associated with local release of cytokines.     

Neuroprotective effects of riluzole and ketamine during transient spinal cord ischemia in the rabbit

BACKGROUND: Massive release of central excitatory neurotransmitters is an important initial step in ischemic neuronal injury, and modification of this process may provide neuroprotection. We studied the protective effects of the voltage-dependent sodium channel antagonist riluzole and the N-methyl-d-aspartate receptor antagonist ketamine on hind limb motor function and histopathologic outcome in an experimental model of spinal cord ischemia. METHODS: Temporary spinal cord ischemia was induced by 29 min of infrarenal balloon occlusion of the aorta in 60 anesthetized New Zealand white rabbits. Animals were randomly assigned to one of four treatment groups (n = 15 each): group C, saline (control); group R, riluzole, 8 mg/kg intravenously; group K, ketamine, 55 mg/kg intravenously; group RK, riluzole and ketamine. After reperfusion, riluzole treatment was continued with intraperitoneal infusions. Normothermia (38 degrees C) was maintained during ischemia, and rectal temperature was assessed before and after intraperitoneal infusions. Neurologic function, according to Tarlov's criteria, was evaluated every 24 h, and infarction volume and the number of eosinophilic neurons and viable motoneurons in the lumbosacral spinal cord was evaluated after 72 h. RESULTS: Neurologic outcome was better in groups R and RK than in groups C and K. All animals in group C (100%) and all animals but one in group K (93%) were paraplegic 72 h after the ischemic insult versus 53% in group R and 67% in group RK (P < 0.01 each). More viable motoneurons were present in groups R and RK than in controls (P < 0.05). CONCLUSIONS: The data indicate that treatment with riluzole can increase the tolerance of spinal cord motoneurons to a period of normothermic ischemia. Intraischemic ketamine did not provide neuroprotection in this model.     

肠缺血再灌注损伤大鼠小肠运动功能的变化与细胞间黏附分子1表达的关系

Objective To evaluate the relationship between the intestinal motility alterations and intercellular adhesion molecule-1 ( ICAM-1 ) expression within the rat intestinal muscularis after ischemia reperfusion. Methods Thirty Wistar rats were divided randomly into IIR, monoclanal anti-ICAM-1 and sham group (n = 10), HR models were established by clamping-releasing the superior mesenteric artery. Gut transit was measured in vivo and intestinal circular muscle contractions were measured in an organ bath. The expression of ICAM-1 mRNA was detected by RT-PCR and immunohistochemisty. Leukocyte infiltration and myeloperoxidase (MPO) activity were quantified in sham and ischemia/reperfusion rats with and without monoclonal anti-ICAM-1 antibody treatment. Results The gut transit of monoclonal anti-ICAM-1 group was improved obviously as compared with IIR group. Circular smooth muscle contractility stimulated by ICAM-1 mRNA expression was 1.69 ± 0. 57 and 1.76 ± 0. 32 within the muscularis; Leukocyte infiltration into the muscularis was (5.6 ±2. 2) c/f and ( 18. 2 ±3. 1 ) c/f. MPO activity was (2. 03 ±0. 56) U/g and (6. 41 ± 1.25 ) U/g respectively. The differences were all statistically significant between IIR and treatment groups ( P ＜ 0. 05 ). The expression of ICAM-1 protein in IIR and anti-ICAM-1 groups was not significantly different (P ＞ 0. 05). Conclusions The up-regulated expression of ICAM-1 after ⅡR injury calls forth local infiltration of PMN within the intestinal muscularis, leading to intestinal motility dysfunction.