不同热缺血时间对猪胰岛获取及功能的影响  

Effect of different warm-ischemia periods on islet isolation and function in pigs

作　　者：张明[1] 杜成友[1] 

机构地区：[1]重庆医科大学附属第一医院肝胆外科,重庆市400016

出　　处：《中国组织工程研究与临床康复》2011年第5期847-850,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：重庆市科委(CSTC2007BB5262);渝卫科教[2006]34号06-2-016~~

摘　　要：背景:目前国内外有关热缺血时间对胰岛移植影响的研究相对较少,更缺乏胰岛移植安全热缺血时限的统一标准。目的:观察不同热缺血时间对胰岛形态、获取及功能的影响。方法:以猪胰腺为观察对象,以不同热缺血时间0,10,20,30,45min采用体、尾部区段胰管内灌注胶原酶Ⅴ和胶原酶Ⅰ混合静置消化,Ficoll400间断密度梯度法纯化胰岛,观察各时间点所得胰岛的形态、活性、胰岛当量数(IEQ)与功能。结果与结论:各时间点均可获得形态完整的胰岛,但随热缺血时间延长形态不完整、破碎胰岛逐渐增加,大于30min时比较明显,45min时很难得到完整的胰岛。0,10,20min胰岛活率大于80%,30min大于60%,45min小于40%。胰岛获得量30,45min与0min之间有显著差异(P<0.05)。葡萄糖刺激胰岛素释放试验30,45min与0min比较有显著差异(P<0.05),且刺激指数<2。结果提示,热缺血时间在20min以内时,分离、纯化后胰岛的形态、产量、活性及功能都无明显的差异,且功能良好;大于30min时,胰岛形态、产量及功能改变显著,且功能低下。BACKGROUND：A few studies address the effect of warm-ischemia time on islet transplantation,moreover,it lacks of unified standards of safe warm ischemia time of islet transplantation. OBJECTIVE：To observe the effect of different warm-ischemia time on islet morphology,yield and function. METHODS：Pig pancreas as the object of observation,collagenase V combined with collagenase I were perfused into pancreatic duct of body and tailer standing to digest,according to warm-ischemia time （WIT） （0,10,20,30,45 min）. Discontinuous Ficoll400 density gradient method was applied to the purification of the islets,and the morphology,activity,equivalent,and function of islets were observed at various times. RESULTS AND CONCLUSION：Integrity islets were obtained at various times. However,with the prolonged time,imperfect morphology,fragmentation of islets increased gradually,more obvious when more than 30 minutes,it was difficult to obtain Integrity islets at 45 minutes. Survival rate of islets was more than 80% at 0,10,20 minutes,survival rate of islets was more than 60% at 30 minutes,and survival rate of islets was less than 40% at 45 minutes. The amount of islets was 30; there were significant differences in the amount of islets between 30,45 minutes and 0 minute （P 0.05）. There were significant differences in glucose-stimulated insulin release test between 30,45 minutes and 0 minute （P 0.05）. The results suggested that there was no significant difference in morphology,yield,activity,and function of islets after isolation and purification when WIT was less than 20 minutes. Moreover,the function of islets was good; the morphology yield,and function of islets significantly change,and hypofunction when WIT was more than 30 minutes.

关 键 词：胰岛 猪 热缺血 时间 胰岛移植 

分 类 号：R318[医药卫生—生物医学工程]