中国临床康复
中國臨床康複
중국림상강복
CHINESE JOURNAL OF CLINICAL REHABILITATION
2006年
24期
175-177
,共3页
背景:近年来亚低温对脑缺血组织的保护作用以及亚低温治疗对脑缺血再灌注后的炎症反应已越来越引起重视.目的:探讨亚低温对大鼠脑缺血区炎性细胞因子的影响,及其再灌注损伤的脑保护机制.设计:以动物为观察对象的随机对照试验.单位:湖北省人民医院.材料:实验于2004-10/2005-02在武汉大学人民医院实验中心进行,选取健康清洁级SD大鼠50只.方法:将SD大鼠50只,先随机选取10只分正常组和假手术组,每组5只;余下的40只随机分为常温脑缺血组和亚低温脑缺血组,每组20只.除正常组5只外,其余大鼠均采用Longa改良线拴法建立可逆的大鼠左侧大脑中动脉线栓模型.假手术组及常温组置于20℃室温,肛温稳定在37℃左右;亚低温组将大鼠置于4℃环境中,全身采用亚低温方法,控制大鼠肛温在(33.0±1.0)℃,缺血结束后立即自然复温,脑缺血3 h后开始再灌注过程.所有造模大鼠均进行神经功能缺失评分(0分为无神经缺损症状;1分为不能伸展对侧前爪;2分为行走时向偏瘫侧转圈;3分为向偏瘫侧倾倒;4分为不能自发行走,意识丧失).主要观察指标:观察缺血脑组织大鼠的肿瘤坏死因子和白细胞介素-1的表达,脑梗死灶体积百分比和神经功能评分.结果:实验过程中有15只大鼠因颅内出血、麻醉意外、神经功能缺失评分不满足要求而被剔除,随机补充15只造模成功大鼠,进入结果分析大鼠保持50只大鼠.①肿瘤坏死因子免疫组化染色阳性细胞数:正常组和假手术组差异无显著性意义[(3.54±1.24,3.71±1.50)个/视野];亚低温脑缺血组明显比常温脑缺血组低[(31.94±7.23,69.20±9.43)个/视野,F=179.16,P＜0.001].②白细胞介素-1免疫组化染色阳性细胞数:正常组和假手术组差异无显著性意义[(3.20±1.34,3.89±2.08)个/视野];亚低温脑缺血组明显比常温脑缺血组低[(28.95±4.97,55.79±7.93)个/视野,F=174.95,P＜0.001].③梗死灶体积百分比:亚低温脑缺血组明显比常温脑缺血组低[(21.06±2.42)%,(30.32±2.71),F=374.87,P＜0.001].④神经功能评分:亚低温脑缺血组明显比常温脑缺血组低[(1.35±0.27)%,(2.04±0.34)%,F=117.17,P＜0.001].结论:①亚低温脑缺血组大鼠脑梗死灶体积较常温脑缺血组明显缩小,提示亚低温对缺血神经元具有保护作用.②正常组和假手术组仅见少许肿瘤坏死因子和白细胞介素1阳性表达,而缺血后再灌注24h脑缺血区即有较多阳性细胞表达,提示脑缺血启动了炎性细胞因子表达,诱发炎症级联反应,从而加重脑缺血再灌注损伤,因此对炎症级联反应的抑制是发挥脑保护作用的重要机制之一.
揹景:近年來亞低溫對腦缺血組織的保護作用以及亞低溫治療對腦缺血再灌註後的炎癥反應已越來越引起重視.目的:探討亞低溫對大鼠腦缺血區炎性細胞因子的影響,及其再灌註損傷的腦保護機製.設計:以動物為觀察對象的隨機對照試驗.單位:湖北省人民醫院.材料:實驗于2004-10/2005-02在武漢大學人民醫院實驗中心進行,選取健康清潔級SD大鼠50隻.方法:將SD大鼠50隻,先隨機選取10隻分正常組和假手術組,每組5隻;餘下的40隻隨機分為常溫腦缺血組和亞低溫腦缺血組,每組20隻.除正常組5隻外,其餘大鼠均採用Longa改良線拴法建立可逆的大鼠左側大腦中動脈線栓模型.假手術組及常溫組置于20℃室溫,肛溫穩定在37℃左右;亞低溫組將大鼠置于4℃環境中,全身採用亞低溫方法,控製大鼠肛溫在(33.0±1.0)℃,缺血結束後立即自然複溫,腦缺血3 h後開始再灌註過程.所有造模大鼠均進行神經功能缺失評分(0分為無神經缺損癥狀;1分為不能伸展對側前爪;2分為行走時嚮偏癱側轉圈;3分為嚮偏癱側傾倒;4分為不能自髮行走,意識喪失).主要觀察指標:觀察缺血腦組織大鼠的腫瘤壞死因子和白細胞介素-1的錶達,腦梗死竈體積百分比和神經功能評分.結果:實驗過程中有15隻大鼠因顱內齣血、痳醉意外、神經功能缺失評分不滿足要求而被剔除,隨機補充15隻造模成功大鼠,進入結果分析大鼠保持50隻大鼠.①腫瘤壞死因子免疫組化染色暘性細胞數:正常組和假手術組差異無顯著性意義[(3.54±1.24,3.71±1.50)箇/視野];亞低溫腦缺血組明顯比常溫腦缺血組低[(31.94±7.23,69.20±9.43)箇/視野,F=179.16,P＜0.001].②白細胞介素-1免疫組化染色暘性細胞數:正常組和假手術組差異無顯著性意義[(3.20±1.34,3.89±2.08)箇/視野];亞低溫腦缺血組明顯比常溫腦缺血組低[(28.95±4.97,55.79±7.93)箇/視野,F=174.95,P＜0.001].③梗死竈體積百分比:亞低溫腦缺血組明顯比常溫腦缺血組低[(21.06±2.42)%,(30.32±2.71),F=374.87,P＜0.001].④神經功能評分:亞低溫腦缺血組明顯比常溫腦缺血組低[(1.35±0.27)%,(2.04±0.34)%,F=117.17,P＜0.001].結論:①亞低溫腦缺血組大鼠腦梗死竈體積較常溫腦缺血組明顯縮小,提示亞低溫對缺血神經元具有保護作用.②正常組和假手術組僅見少許腫瘤壞死因子和白細胞介素1暘性錶達,而缺血後再灌註24h腦缺血區即有較多暘性細胞錶達,提示腦缺血啟動瞭炎性細胞因子錶達,誘髮炎癥級聯反應,從而加重腦缺血再灌註損傷,因此對炎癥級聯反應的抑製是髮揮腦保護作用的重要機製之一.
배경:근년래아저온대뇌결혈조직적보호작용이급아저온치료대뇌결혈재관주후적염증반응이월래월인기중시.목적:탐토아저온대대서뇌결혈구염성세포인자적영향,급기재관주손상적뇌보호궤제.설계:이동물위관찰대상적수궤대조시험.단위:호북성인민의원.재료:실험우2004-10/2005-02재무한대학인민의원실험중심진행,선취건강청길급SD대서50지.방법:장SD대서50지,선수궤선취10지분정상조화가수술조,매조5지;여하적40지수궤분위상온뇌결혈조화아저온뇌결혈조,매조20지.제정상조5지외,기여대서균채용Longa개량선전법건립가역적대서좌측대뇌중동맥선전모형.가수술조급상온조치우20℃실온,항온은정재37℃좌우;아저온조장대서치우4℃배경중,전신채용아저온방법,공제대서항온재(33.0±1.0)℃,결혈결속후립즉자연복온,뇌결혈3 h후개시재관주과정.소유조모대서균진행신경공능결실평분(0분위무신경결손증상;1분위불능신전대측전조;2분위행주시향편탄측전권;3분위향편탄측경도;4분위불능자발행주,의식상실).주요관찰지표:관찰결혈뇌조직대서적종류배사인자화백세포개소-1적표체,뇌경사조체적백분비화신경공능평분.결과:실험과정중유15지대서인로내출혈、마취의외、신경공능결실평분불만족요구이피척제,수궤보충15지조모성공대서,진입결과분석대서보지50지대서.①종류배사인자면역조화염색양성세포수:정상조화가수술조차이무현저성의의[(3.54±1.24,3.71±1.50)개/시야];아저온뇌결혈조명현비상온뇌결혈조저[(31.94±7.23,69.20±9.43)개/시야,F=179.16,P＜0.001].②백세포개소-1면역조화염색양성세포수:정상조화가수술조차이무현저성의의[(3.20±1.34,3.89±2.08)개/시야];아저온뇌결혈조명현비상온뇌결혈조저[(28.95±4.97,55.79±7.93)개/시야,F=174.95,P＜0.001].③경사조체적백분비:아저온뇌결혈조명현비상온뇌결혈조저[(21.06±2.42)%,(30.32±2.71),F=374.87,P＜0.001].④신경공능평분:아저온뇌결혈조명현비상온뇌결혈조저[(1.35±0.27)%,(2.04±0.34)%,F=117.17,P＜0.001].결론:①아저온뇌결혈조대서뇌경사조체적교상온뇌결혈조명현축소,제시아저온대결혈신경원구유보호작용.②정상조화가수술조부견소허종류배사인자화백세포개소1양성표체,이결혈후재관주24h뇌결혈구즉유교다양성세포표체,제시뇌결혈계동료염성세포인자표체,유발염증급련반응,종이가중뇌결혈재관주손상,인차대염증급련반응적억제시발휘뇌보호작용적중요궤제지일.
BACKGROUND: At present, more and more people pay attention to the protection of sub-hypothermia on cerebral ischemic tissue and the effect of sub-hypothermia treatment on inflammatory reaction after cerebral ischemia-reperfusion.OBJECTIVE: To investigate the effect of sub-hypothermia on inflammatory cytokine in cerebral ischemic area of rats and protection on cerebral reperfusion injury.DESIGN: Randomized controlled study on the basis of animals.SETTLNG: People's Hospital of Hubei Province.MATERIALS: The experiment was completed at Laboratory Center of Renmin Hospital of Wuhan University from October 2004 to February 2005. Totally 50 healthy SD rats of clean grade were selected in this study.METHODS: Ten SD rats were divided randomly into normal group and sham operation group with 5 in each group. Other 40 rats were divided randomly into normal temperature cerebral ischemia group and sub-hypothermia cerebral ischemia group with 20 in each group. Rats except 5 in normal group were used to establish reversible ligation model of left middle cerebral artery (MCA) with Longa ligation method. Rats in sham operation group and normal temperature group were put in 20 ℃ room, and anus temperature was maintained at 37 ℃; rats in sub-hypothermia group were put in 4 ℃ room and anus temperature was maintained at (33.0±1.0) ℃.Rats in sub-hypothermia group were treated with sub-hypothermia on whole body, after ischemia, temperature was changed normally, and reperfusion was started 3 hours after cerebral ischemia. All modeling rats were scored with neurological defect: 0 point: none of neurological defect; 1 point: unable to unfold bilateral anterior claws; 2 points: cycling to hemiplegia side during walking; 3 points: falling to hemiplegia side; 4 points: unable to self-walk and with unconsciousness.MAIN OUTCOME MEASURES: Expression of tumor necrosis factor (TNF) andinterleukin-1 in cerebral tissue of ischemic rats, percentage of volume of cerebral infarction,and neurological functional scores.RESULTS: Fifteen rats were excluded because of intracranial hemorrhage,anesthetic accident and unqualified neurological functional scores. Other 15 rats with successful modeling were supplied randomly, and totally 50rats entered the final analysis. ① Number of TNF positive cell in immunohistochemical staining: There were no significant differences between normal group and sham operation group [(3.54±1.24, 3.71±1.50) /sight]; but numbers in sub-hypothermia group were lower than those in normal temperature group [(31.94±7.23, 69.20±9.43)/sight, F=179.16, P ＜ 0.001]. ②Numbers of interleukin-1 positive cells in immunohistochemical staining:There were not significant differences between normal group and sham operation group [(3.20±1.34, 3.89±2.08) /sight]; but numbers in sub-hypothermia group were lower than those in normal temperature group [(28.95±4.97, 55.79±7.93)/sight, F=174.95, P ＜ 0.001]. ③ Volume percentage of infarct focus: Percentage in sub-hypothermia group was lower than that in normal temperature group [(21.06±2.42)%, 30.32±2.71], F =374.87, P ＜ 0.001]. ④ Neurological functional scores: Percentage in subhypothermia group was lower than that in normal temperature group [(1.35±0.27)%, (2.04±0.34)%, F=117.17, P ＜ 0.001].CONCLUSION: ① Volume of cerebral infarction focus is lower in subhypothermia group than that in normal temperature group, this suggests that sub-hypothermia can protect ischemic neurons. ② Positive expressions of TNF and interleukin-1 are observed rarely in normal group and sham operation group, but more expression of positive cells are observed in cerebral ischemic area with 24-hour reperfusion after ischemia, this suggests that cerebral ischemia starts expression of inflammatory cytokine, evoke inflammatory cascade reaction, deteriorate cerebral ischemia-reperfusion injury. Therefore, inhibition of inflammatory cascade reaction is one of important mechanisms of protecting brain.
