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30 min 7只,阻断血流60 min 7只;缺血再灌注组:根据脊髓缺血30 min后再灌注30,60min,2,4,6,9,12,24 h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1 mRNA和白细胞介素1β mRNA表达量.主要观察指标:白细胞介素1β mRNA表达、白细胞介素1多肽活性、细胞间黏附分子1 mRNA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1β mRNA(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=3.751 7,11.852 6,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=2.798 8,9.962 7,P＜0.01].③缺血再灌注组细胞间黏附分子1 mRNA(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=0.327 0,6.127 4,P＜0.01].④缺血再灌注4,6,12 h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=1.410 3,9.119 3,P＜0.01].⑤缺血再灌注12 h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),(750±1.67),(6.67±1.00)nkat/g,t=3.012 2,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.
揹景:目前國外有探討細胞因子和黏附分子在缺血再灌註損傷時的錶達,但均未涉及內源性細胞因子和微血管內皮細胞錶麵黏附分子在損傷後相關性的研究,對內源性白細胞介素1在損傷中的錶達也僅限于mRNA水平.目的:探討細胞間黏附分子1及其調節因子白細胞介素1β的錶達在脊髓缺血再灌註損傷中作用機製.設計:隨機分組設計、動物實驗.單位:吉林大學體育學院運動醫學繫.材料:實驗于2003-03/2004-01在吉林大學中日聯誼醫院中心實驗室完成.選擇健康雄性Wistar大鼠77隻,隨機數字錶法分為正常對照組7隻,單純缺血組14隻和缺血再灌註組56隻.單純缺血組:阻斷血流30 min 7隻,阻斷血流60 min 7隻;缺血再灌註組:根據脊髓缺血30 min後再灌註30,60min,2,4,6,9,12,24 h又分為8箇時相點,每箇時相點7隻.方法:採用Zivin法複製脊髓缺血再灌註損傷動物模型.採用反轉錄-聚閤酶鏈反應、免疫組織化學及免疫熒光激光共聚焦掃描顯微鏡技術檢測脊髓缺血再灌註損傷後血管內皮細胞間黏附分子1 mRNA和白細胞介素1β mRNA錶達量.主要觀察指標:白細胞介素1β mRNA錶達、白細胞介素1多肽活性、細胞間黏附分子1 mRNA和蛋白的錶達、髓過氧化物酶活性.結果:納入動物77隻,均進入結果分析.①缺血再灌註組白細胞介素1β mRNA(A值)錶達量明顯高于單純缺血組和正常對照組,差異顯著(分彆為1.07±0.33,0.60±0.22,0.57±0.12,t=3.751 7,11.852 6,P＜0.01).②缺血再灌註組白細胞介素1多肽活性(A值)明顯高于單純缺血組和正常對照組,差異顯著[分彆為(33.7±3.2),(23.8±4.5),(23.1±2.1),t=2.798 8,9.962 7,P＜0.01].③缺血再灌註組細胞間黏附分子1 mRNA(A值)明顯高于單純缺血組和正常對照組,差異顯著[分彆為0.94±0.12,0.52±0.11,0.51±0.10,t=0.327 0,6.127 4,P＜0.01].④缺血再灌註4,6,12 h各組細胞間黏附分子1蛋白的錶達明顯高于單純缺血組和正常對照組,差異顯著[分彆為(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缺血再灌註12 h組髓過氧化物酶活性明顯高于單純缺血組和正常對照組,差異顯著[分彆為(15,00±2.00),(750±1.67),(6.67±1.00)nkat/g,t=3.012 2,P＜0.01].結論:再灌註損傷後脊髓內炎癥反應是導緻血脊髓屏障損害的重要分子基礎,在繼髮性脊髓損傷過程中起到重要作用.
배경:목전국외유탐토세포인자화점부분자재결혈재관주손상시적표체,단균미섭급내원성세포인자화미혈관내피세포표면점부분자재손상후상관성적연구,대내원성백세포개소1재손상중적표체야부한우mRNA수평.목적:탐토세포간점부분자1급기조절인자백세포개소1β적표체재척수결혈재관주손상중작용궤제.설계:수궤분조설계、동물실험.단위:길림대학체육학원운동의학계.재료:실험우2003-03/2004-01재길림대학중일련의의원중심실험실완성.선택건강웅성Wistar대서77지,수궤수자표법분위정상대조조7지,단순결혈조14지화결혈재관주조56지.단순결혈조:조단혈류30 min 7지,조단혈류60 min 7지;결혈재관주조:근거척수결혈30 min후재관주30,60min,2,4,6,9,12,24 h우분위8개시상점,매개시상점7지.방법:채용Zivin법복제척수결혈재관주손상동물모형.채용반전록-취합매련반응、면역조직화학급면역형광격광공취초소묘현미경기술검측척수결혈재관주손상후혈관내피세포간점부분자1 mRNA화백세포개소1β mRNA표체량.주요관찰지표:백세포개소1β mRNA표체、백세포개소1다태활성、세포간점부분자1 mRNA화단백적표체、수과양화물매활성.결과:납입동물77지,균진입결과분석.①결혈재관주조백세포개소1β mRNA(A치)표체량명현고우단순결혈조화정상대조조,차이현저(분별위1.07±0.33,0.60±0.22,0.57±0.12,t=3.751 7,11.852 6,P＜0.01).②결혈재관주조백세포개소1다태활성(A치)명현고우단순결혈조화정상대조조,차이현저[분별위(33.7±3.2),(23.8±4.5),(23.1±2.1),t=2.798 8,9.962 7,P＜0.01].③결혈재관주조세포간점부분자1 mRNA(A치)명현고우단순결혈조화정상대조조,차이현저[분별위0.94±0.12,0.52±0.11,0.51±0.10,t=0.327 0,6.127 4,P＜0.01].④결혈재관주4,6,12 h각조세포간점부분자1단백적표체명현고우단순결혈조화정상대조조,차이현저[분별위(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].⑤결혈재관주12 h조수과양화물매활성명현고우단순결혈조화정상대조조,차이현저[분별위(15,00±2.00),(750±1.67),(6.67±1.00)nkat/g,t=3.012 2,P＜0.01].결론:재관주손상후척수내염증반응시도치혈척수병장손해적중요분자기출,재계발성척수손상과정중기도중요작용.
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 UniversityMATERIALS: 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 transcriptionpolymerase 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).RFSULTS: 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 ischemiareperfusion 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.
