【作者】 康义华；

【导师】 陈嘉勇；

【作者基本信息】 昆明医学院 ， 外科学， 2010， 硕士

【摘要】 目的通过大鼠小肠缺血再灌注(Ischemia-reperfusion I/R)对小肠粘膜的损伤程度,促炎因子和抗炎因子的释放方面研究依达拉奉(Edaravone 3-甲基-i-苯基-2-N比啉唑-5-酮)对小肠I/R损伤的保护作用,从而为临床防治小肠I/R损伤提供一个新的治疗思路。方法SPF级健康成年SD大鼠60只,体重在280g-320g之间,均为雄性,随机分为对照组、I/R组和依达拉奉组3组(n=20),每组根据再灌注时间不同再分为2小时组和4小时组两个亚组(n=10)。于实验前一天禁食,自由饮水,实验前2小时行含绿色荧光蛋白大肠杆菌灌胃。实验中对照组开腹后仅分离肠系膜上动脉(SMA)后关腹;I/R组分离SMA后以无创动脉夹夹闭,45min后松开动脉夹,分别于再灌注2小时、4小时时采集标本;依达拉奉组夹闭SMA45min后行再灌注,于再灌注前5min经大鼠尾静脉按3 mg/kg剂量注入依达拉奉,分别于再灌注2小时、4小时时采集标本。小肠粘膜组织HE染色后病理学检测,对门静脉血、肝脏组织及脾脏组织行细菌学培养,ELISA法检测大鼠血清IL-1β、IL-6、IL-10和TNF-α的含量。结果(1)肠粘膜病理变化:对照组大鼠小肠绒毛完整,排列整齐,炎性细胞浸润少,无明显出血及脱落。I/R组大鼠在再灌注2小时和4小时小肠绒毛上皮顶端与固有层分离,固有层出血和溃疡,大量炎性细胞浸润,粘膜绒毛坏死脱落。与I/R组相比,依达拉奉组在再灌注2小时和4小时肠粘膜绒毛损伤减轻,坏死,脱落减少。依达拉奉组病理损伤评分低于I/R组(p＜0.05)。(2)细胞因子变化:I/R组和依达拉奉组再灌注2小时和4小时大鼠血清中炎性细胞因子IL-1β、IL-6、TNF-α含量明显高于对照组(P＜0.05),I/R组高于依达拉奉组(p＜0.05),I/R组和依达拉奉组再灌注2小时和4小时时大鼠血清中抗炎因子IL-10含量明显高于对照组(p＜0.05),依达拉奉组高于I/R组(p＜0.05)。(3)细菌移位情况:I/R组、依达拉奉组再灌注2小时、4小时后大鼠门静脉、肝脏、脾脏的细菌移位明显高于正常对照组(p＜0.05),I/R组高于依达拉奉组(p＜0.05)。结论依达拉奉对大鼠小肠I/R具有保护作用。(1)依达拉奉能有效减轻大鼠小肠I/R损伤小肠粘膜绒毛的坏死程度,减少绒毛脱落,保护大鼠小肠粘膜结构的完整性和正常生理功能。(2)依达拉奉能明显减少大鼠小肠I/R损伤促炎细胞因子IL-1β、IL-6、TNF-α的含量和促进抗炎细胞因子IL-10的释放。(3)依达拉奉可明显降低大鼠小肠I/R损伤后肠道细菌移位。更多还原

【Abstract】 [Objective]To study the protective effects of edaravone against ischemia-reperfusion injury of small intestine in rat intestinal mucosa damage,inflammatory factorand anti-inflammatory factor release in order to find a way to protect clinical small intestine from I/R injury.[Methods]Sixty SD rats weighted about 280g-320g were divided into three groups randomly:control group,I/R group,edaravone group(n=20).Each group was divided into two subgroups according to time of reperfusion (n=10),The control group:the rats underwent a 3cm medium length-wise laparotomy,exposure of the small intestine,identification and dissection of the SMA and then,closure abdominal cavity.The I/R group:the rats were operated by occlusion of SMA for 45 min,and then,same with control group. The edaravone group:the rats were operated by occlusion of SMA for 45 min, 5 min before reperfusion,inject edaravone through tail vein(3mg/kg),and then,same with I/R group.At the end of the operation,the rats were sacrificed,and dissected 10cm segment of ileum to determination of intestines mucous membrane’s injury.The serum of TNF-α,IL-1β, IL-6,IL-10 are measured by ELISA.Cut tissues from liver and spleen at the same weight and portal vein blood to detect bacteria.[Results](1)The changes of intestinal mucosa:In I/R group,the intestinal mucosa presented dilated and exposed capillaries and denuded villi,some of villi hemorrhage,ulceration.The lamina propria was edema and infiltrated with inflammatory cells.The intestinal mucosal structure maintain intact with lifting of the epithelial layer from the lamina propria and moderate extension of the subepithelial space in edaravone group.In edaravone group,histopathologic scores were significantly lower than that of I/R group.(2) Compared with Contraol group,the serum contents of TNF-α,IL-1β,IL-6 in I/R group,edaravone group are obviously higher, and the edaravone group is lower than that of I/R group.The contents of IL-10 in other two groups are lower than that of control group,and edaravone group are higher than that of I/R group.(3)Compared with control group, the bacteria transposition in I/R group,edaravone group is obviously higher,and the edaravone group is lower than that of I/R group.[Conclusion]Edaravone can protects against small intestinal ischemia-reperfusion injury in rats.(1)Edaravone makes small intestinal mucosal damage degree alleviated in rat I/R injury.(2) Edaravone can reduce the contents of inflammatory TNF-α,IL-1β,IL-6 and promote the contents of anti-inflammatory medium Ih-10 in rat I/R injury.(3) Edaravone can reduce the bacterias transposition of small intestine in rat I/R ingury.更多还原