摘要
背景:细胞凋亡在肠缺血再灌注损伤中起关键性作用,细胞凋亡的发生与凋亡抑制基因Bcl-2和凋亡促进基因Bax蛋白表达程度密切相关。目的:探讨亚低温缺血预处理对大鼠肠组织缺血再灌注后凋亡相关基因Bcl-2和Bax蛋白表达的影响。设计:随机对照实验。单位:湖北省咸宁学院医学院外科学教研室与生物化学教研室。材料:实验于2002—04/12在咸宁学院医学院中心实验室完成。选用Wistar健康雄性大鼠32只,术前禁食24h,自由饮水。随机分为假手术对照组、缺血再灌注组、缺血预处理组、亚低温预处理组,8只/组。方法:除假手术对照组外,其余3组建立缺血再灌注模型。①假手术对照组仅暴露肠系膜上动脉而不夹闭,共2h,结束实验后取材;缺血再灌注组夹闭肠系膜上动脉60min后松夹60min再取材;缺血预处理组夹闭肠系膜上动脉5min和松夹5min作为预处理,余同缺血再灌注组;亚低温预处理组实施缺血预处理前在小肠周围充填碎冰造成小肠亚低温(33～35℃),余同缺血预处理组。②各组大鼠取中段小肠4cm,分为两段,分别置于甲醛和戊二醛中固定,制作电镜标本。免疫组化后显微镜下用图像分析系统检测各组吸光度值,每组选10个视野进行测量,计算平均值,肠黏膜损伤按Chiu标准进行分级。0级:正常黏膜绒毛;Ⅰ级:上皮下间隙增大,通常在绒毛的尖端,常伴有毛细血管淤血;Ⅱ级:上皮下间隙扩张伴随上皮层同固有层中度分离;Ⅲ级:绒毛两侧上皮层大量的同固有层分离,部分绒毛顶端破损;Ⅳ级绒毛破损伴随固有层毛细血管暴露,可能观察到固有层的细胞成分增多;Ⅴ级:固有层破坏和不完整、出血和溃疡。主要观察指标:①各组肠缺血再灌注后Bcl-2与Bax蛋白吸光度值的变化。②各组肠黏膜损伤分级。③各组肠黏膜组织学变化。结果:32只大鼠全部进入结果分析,中途无脱落。①各组肠缺血再�
BACKGROUND： Cellular apoptosis plays a key role in intestinal ischemia-reperfusion injury and is closely related to the expression level of apoptosis-inhibiting Bcl-2 and apoptosis-promoting Bax. OBJECTIVE： To probe into the effect of sub-hypothermia ischemie pretreatment on the expression of apoptosis-related Bcl-2 and Bax protein after intestinal ischemia-reperfusion in rats. DESIGN： Randomized controlled trial. SETTING： Department of Surgery and Department of Biochemistry, Medical School of Xianning College of Hubei Province. MATERIALS： The experiment was conducted in the Central Laboratory of the Medical School of Xianning College from April to December 2002. Totally 32 male healthy Wistar rats were used. The animals were fasted for 24 hours before operation with free access to water. Then they were randomly divided into sham-operation control group, ischemia-reperfusion group, ischemia-pretreatment group, and sub-hypothermia pre-treatment group with 8 rats in each group. METHODS： lschemia-reperfusion model was established in all the groups except sham-operation group.①Superior mesenteric artery was exposed for 2 hours without occlusion in the rats of sham-operation group; the material was selected after the experiment. Superior mesenteric artery in ischemia-reperfusion group was occluded for 60 minutes; the material was selected after stopping occlusion for another 60 minutes. Superior mesenteric artery in ischemia-pretreatment group was occluded for 5 minutes and stopped occlusion for 5 minutes, and the other procedure was the same as that in ischemia-reperfusion group. In sub-hypothermia pretreatment group, pieces of fractional ice filled around the small intestine for sub-hypothermia （33-35℃） before performing ischemia pretreatment, then followed by the same procedure as that in ischemia-pretreatment group. ②The middle part （4 cm） of the rats＇ small intestine was chosen in each group. Each small intestine was divided into 2 segments, whicb were put in formaldehyde and glutaric
出处
《中国临床康复》
CSCD
北大核心
2005年第30期239-241,共3页
Chinese Journal of Clinical Rehabilitation
基金
湖北省教育厅基金重点资助课题(2002A04005)~~
