肠缺血再灌注时肿瘤坏死因子免疫组织化学研究

Immunohistochemical studies of tumour necrosis factor during gut ischemia/reperfusion injury in rat pups

目的 探讨肠缺血 /再灌注时肿瘤坏死因子 (tumornecrosisfactorα,TNF α)的来源及作用。方法 ① 4 8只雄性Wistar大白鼠体重 (130± 15 ) g ,分成 6组 (n =8) ,假手术组 ,缺血 30min组 ;缺血 30min ,再灌注组 ;缺血 30min ,再灌注 6 0min组 ;缺血 30min ,再灌注 90min组 ;缺血 12 0min组 ;②利用免疫组织化学技术检测肠、肝组织TNF α蛋白表达和分布。③对肠组织进行HE染色。结果 ①肠粘膜免疫组化于缺血 130组 ;缺血 30min ,再灌注 6 0min组 ;缺血 30min ,再灌注 90min组时可见TNF α强阳性物质 ,缺血 30min ,再灌注 90min组 ;可见极强阳性物质 ;②缺血 30min ,再灌注 30min组 ;缺血 30min ,再灌注 6 0min组肝组织内可见阳性物质 ,缺血 30min ,再灌注 90min组 ;为强阳性物质 ;③肠组织HE染色显示I30’G肠粘膜轻度受损 ,缺血 30min ,用灌注 30min组 ;缺血 30min ,再灌注 6 0min组 ;缺血 30min ,再灌注 90min组肠粘膜中度受损 ,再灌注 12 0min组肠粘膜重度受损。结论 幼鼠肠缺血 /再灌注后TNF α在肠、肝内均有阳性反应 ,但肠粘膜内TNF α较肝内明显 ,因此推论幼鼠肠缺血 /再灌注损伤早期TNF α可能主要来源于肠组织 ,并且介导幼鼠肠缺血

Objective To investigate the source and effects of tumour necrosis factor (TNF α) in the model of rat pups after gut ischemia/reperfusion (I/R) injury.Methods 1. Forty eight male Wistar rat pups (wt.130±15?g) were randomly divided into six groups (8 for each group): 1) sham operation group (SG), 2) ischemia 30'group (I30'G), 3) ischemia 30'/reperfusion30'group (I30'/R30'G), 4) ischemia30'/ reperfusion 60'group (I30'/R60'G), 5) ischemia30'/reperfusion90' group ( I30'/R90'G), 6) ischemia 120' group (I120'G). 2.Using SP immunohistochemistry, the expression and distribution of TNF α in the gut and liver were assessed. 3. The pathologic changes of gut were observed by H. & E staining. Results 1.TNF α positive material stained strongly in gut mucosal cells in I30'G ,I30'/R60'G ,I30'/R90'G; very strongly in I30'/R30'G and 120'G.2.TNF α appeared weak in I30'G in liver; moderately in I30'/R30'G and I30'/R60'G;stongly in I30'/90'G and I120'G.3. The gut mucosal villi were weakly damaged with edema in I30'G; more seriously with necrosis or hemorrhage in I30'/R30'G and I120'G.Conclusions TNF α appears in the cytoplasm of gut and liver in the rat pup model after gut I/R injury. Compared with in the liver tissue, TNF α stains more strongly in the gut. This suggests the gut might be the major source of TNF α, which mediates gut I/R injury.