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脊髓再灌注损伤后细胞间黏附分子1及白细胞介素1β表达的变化(英文)
Changes in the expression of intercellular adhesion molecular 1 and interleukin-1 beta following spinal reperfusion injury
【摘要】 背景:目前国外有探讨细胞因子和黏附分子在缺血再灌注损伤时的表达,但均未涉及内源性细胞因子和微血管内皮细胞表面黏附分子在损伤后相关性的研究,对内源性白细胞介素1在损伤中的表达也仅限于mRNA水平。目的:探讨细胞间黏附分子1及其调节因子白细胞介素1β的表达在脊髓缺血再灌注损伤中作用机制。设计:随机分组设计、动物实验。单位:吉林大学体育学院运动医学系。材料:实验于2003-03/2004-01在吉林大学中日联谊医院中心实验室完成。选择健康雄性Wistar大鼠77只,随机数字表法分为正常对照组7只,单纯缺血组14只和缺血再灌注组56只。单纯缺血组:阻断血流30min7只,阻断血流60min7只;缺血再灌注组:根据脊髓缺血30min后再灌注30,60min,2,4,6,9,12,24h又分为8个时相点,每个时相点7只。方法:采用Zivin法复制脊髓缺血再灌注损伤动物模型。采用反转录-聚合酶链反应、免疫组织化学及免疫荧光激光共聚焦扫描显微镜技术检测脊髓缺血再灌注损伤后血管内皮细胞间黏附分子1mRNA和白细胞介素1βmRNA表达量。主要观察指标:白细胞介素1βmRNA表达、白细胞介素1多肽活性、细胞间黏附分子1mRNA和蛋白的表达、髓过氧化物酶活性。结果:纳入动物77只,均进入结果分析。①缺血再灌注组白细胞介素1βmRNA(A值)表达量明显高于单纯缺血组和正常对照组,差异显著(分别为1.07±0.33,0.60±0.22,0.57±0.12,t=3.7517,11.8526,P<0.01)。②缺血再灌注组白细胞介素1多肽活性(A值)明显高于单纯缺血组和正常对照组,差异显著[分别为(33.7±3.2),(23.8±4.5),(23.1±2.1),t=2.7988,9.9627,P<0.01]。③缺血再灌注组细胞间黏附分子1mRNA(A值)明显高于单纯缺血组和正常对照组,差异显著[分别为0.94±0.12,0.52±0.11,0.51±0.10,t=0.3270,6.1274,P<0.01]。④缺血再灌注4,6,12h各组细胞间黏附分子1蛋白的表达明显高于单纯缺血组和正常对照组,差异显著[分别为(316.90±26.00),(361.40±18.00),(406.00±23.00),(164.21±2.00),(180.00±32.00)μg/L,t=1.4103,9.1193,P<0.01]。⑤缺血再灌注12h组髓过氧化物酶活性明显高于单纯缺血组和正常对照组,差异显著[分别为(15.00±2.00),(7.50±1.67),(6.67±1.00)nkat/g,t=3.0122,P<0.01]。结论:再灌注损伤后脊髓内炎症反应是导致血脊髓屏障损害的重要分子基础,在继发性脊髓损伤过程中起到重要作用。
【Abstract】 BACKGROUND: At present,there are investigations on the expression of cytokines and adhesion molecular in ischemia-reperfusion injury at abroad,but they do not involve in the relative studies on endogenous cytokines and adhesion molecular on microvascular endothelial surface following injury. The expression of endogenous interleukin-1(IL-1) is limited only at mRNA level. OBJECTIVE: To prove into the mechanism of the expression of intercellular adhesion molecular 1 and its regulation factor IL-1 in spinal ischemia-reperfusion injury. DESIGN: A randomized grouping design,animal experiment. SETTING: Department of Sports Medicine,College of Physical Education Affiliated to Jilin University MATERIALS: This experiment was carried out at the Central Laboratory,China-Japan Union Hospital,Jilin University between March 2003 and January 2004. Totally 77 healthy male Wistar rats were randomly divided into normal control group (n=7),simple ischemia group (n=14) and ischemia-reperfusion group (n=56). Among the rats in the simple-ischemia group,7 rats suffered from blood flow block for 30 minutes and 7 rats for 60 minutes; Rats in the ischemia-reperfusion group were assigned into 8 subgroups according to 8 time phases,respectively at reperfusion for 30,60 minutes,2,4,6,9,12 and 24 hours following spinal ischemia,with 7 rats at each time phase. METHODS: Spinal ischemia-reperfusion injury animal models were created with Zivin method. The expressions of vascular endothelial intercellular adhesion molecule-1(ICAM-1) mRNA and IL-1β mRNA following spinal ischemia-reperfusion injury were detected with reverse transcription-polymerase chain reaction,immunohistochemistry and immunofluorescent confocal laser scanning microscope technique. MAIN OUTCOME MEASURES: The expression of IL-1β mRNA,activity of IL-1 polypeptide,expression of ICAM-1 mRNA and protein and activity of myeloperoxidase (MPO). RESULTS: Totally 77 animals were enrolled and all of them entered the stage of result analysis. ① The expression of IL-1β mRNA (A value) was significantly higher in the ischemia-reperfusion group than in the simple ischemia group and normal control group,with significant difference (respectively 1.07±0.33,0.60±0.22,0.57±0.12,t=3.751 7,11.852 6,P < 0.01).② Activity of IL-1 polypeptide (A value )was significantly higher in the ischemia-reperfusion group than in the simple ischemia group and normal control group,with significant difference [respectively (33.7±3.2),(23.8±4.5),(23.1±2.1),t=2.798 8,9.962 7,P < 0.01]. ③ ICAM-1 mRNA (A value)was significantly higher in ischemia-reperfusion group than in simple ischemia group and normal control group,with significant difference [respectively 0.94±0.12,0.52±0.11,0.51±0.10,t=0.327 0,6.127 4,P < 0.01]. ④The expression of ICAM-1 protein was significantly higher at ischemia-reperfusion for 4,6 and 12 hours than in simple ischemia group and normal control group,with significant difference [Respectively (316.90±26.00),(361.40±18.00),(406.00±23.00),(164.21±2.00),(180.00±32.00) μg/L,t=1.410 3,9.119 3,P < 0.01]. ⑤ The activity of MPO was significantly higher at ischemia-reperfusion for 12 hours than in simple ischemia group and normal control group,with significant difference [respectively(15.00±2.00),(7.50±1.67),(6.67±1.00) nkat/g,t=3.012 2,P < 0.01]. CONCLUSION: Following reperfusion injury,inflammatory reaction in spinal cord is important molecular basis for causing blood spinal barrier impairment,and plays an important role in the process of secondary spinal cord injury.
- 【文献出处】 中国临床康复 ,Chinese Journal of Clinical Rehabilitation , 编辑部邮箱 ,2006年40期
- 【分类号】R651.2
- 【被引频次】3
- 【下载频次】131