CAJ | 학술논문

目的探讨纤维连接蛋白连接片段-1(CS1)肽段对大鼠肝移植缺血再灌注损伤的影响及其机制.方法用Wistar大鼠作为供、受鼠,制作肝移植模型.CS1组供鼠分别于术前3d、取肝时和移植前注射CS1肽段,供肝获取后于UW液中保存18h,肝移植术后3d每天注射CS1肽段；对照组用随机肽段替代CS1,其他操作同CS1组.术后6、24、72 h检测受鼠血清转氨酶水平和肝组织病理改变.免疫组织化学染色显示肝脏中的炎症细胞和肝窦内皮细胞.多聚酶链反应检测肝组织中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和血管内皮细胞生长因子(VEGF) mRNA的表达水平.结果术后24 h,对照组坏死肝组织面积约占总面积的(25.7±5.4)％,而CS1组为(12.6±3.6)％(P＜0.05).对照组血清转氨酶水平也低于对照组(P＜0.05).术后CS1组移植肝中枯否细胞和中性粒细胞数目均少于对照组(P＜0.05).两组冷缺血18h的供肝中,肝窦内皮细胞均失去正常形态,仅少数内皮细胞特异性标志物阳性.术后72 h,CS1组肝窦内皮细胞基本恢复正常形态,SE-1表达恢复,而对照组肝窦内皮细胞恢复欠佳.术后6和24 h时,CS1组肝组织中TNF-α mRNA的水平低于对照组(P＜0.0)5)；术后24 h时,对照组VEGF mRNA的表达高于CS1组(P＜0.05)；两组IL-1βmRNA水平的差异无统计学意义(P＞0.05).结论用CS1肽段处理供、受鼠可以减少炎症因子mRNA的表达,保护肝窦内皮细胞,减轻移植肝缺血再灌注损伤.
목적 탐토섬유련접단백련접편단-1(CS1)태단대대서간이식결혈재관주손상적영향급기궤제.방법 용Wistar대서작위공、수서,제작간이식모형.CS1조공서분별우술전3d、취간시화이식전주사CS1태단,공간획취후우UW액중보존18h,간이식술후3d매천주사CS1태단；대조조용수궤태단체대CS1,기타조작동CS1조.술후6、24、72 h검측수서혈청전안매수평화간조직병리개변.면역조직화학염색현시간장중적염증세포화간두내피세포.다취매련반응검측간조직중종류배사인자-α(TNF-α)、백세포개소-1β(IL-1β)화혈관내피세포생장인자(VEGF) mRNA적표체수평.결과 술후24 h,대조조배사간조직면적약점총면적적(25.7±5.4)％,이CS1조위(12.6±3.6)％(P＜0.05).대조조혈청전안매수평야저우대조조(P＜0.05).술후CS1조이식간중고부세포화중성립세포수목균소우대조조(P＜0.05).량조랭결혈18h적공간중,간두내피세포균실거정상형태,부소수내피세포특이성표지물양성.술후72 h,CS1조간두내피세포기본회복정상형태,SE-1표체회복,이대조조간두내피세포회복흠가.술후6화24 h시,CS1조간조직중TNF-α mRNA적수평저우대조조(P＜0.0)5)；술후24 h시,대조조VEGF mRNA적표체고우CS1조(P＜0.05)；량조IL-1βmRNA수평적차이무통계학의의(P＞0.05).결론 용CS1태단처리공、수서가이감소염증인자mRNA적표체,보호간두내피세포,감경이식간결혈재관주손상.
Objective To examine the effect of fibronectin connecting segment-1 (CS1) peptidefacilitated blockade of inflammatory cells-fibronectin adhesion on a rat liver transplantation model of prolonged ex vivo cold ischemia.Methods A model of liver transplantation in Wistar→Wistar rat was established.The donors of the CS1 treatment group received CS1 peptides through the tail vein for 3 days before operation.Another two doses of CS1 peptides were administered into the liver intraportally during procurement and before transplantion.Recipients received an additional 3-day course of CS1 peptides after transplantation.Rats in control group received scrambled peptides.Rats were sacrificed at 6,24 and 72 h after transplantion,and plasma transaminase activity and hepatic pathological changes were studied.The inflammatory cells and liver sinusoidal endothelial cells were visualized histochemically.Real-time PCR was used to detect tumor necrosis factor-α (TNF-α),interleukin-1β (IL-1β) and vascular endothelial growth factor (VEGF) mRNA expression in the liver.Results The plasma transaminase activity and hepatic necrosis areas in CS1 treatment group were significantly lower than in control group (P＜0.05).CS1 peptides treatment significantly decreased the number of Kupffer cells after transplantation and greatly inhibited the recruitment of neutrophils to the graft liver as compared with control group (P＜0.05).After prolonged cold ischemia,only a few hepatic endothelial cells exhibited positive staining of hepatic sinusoidal endothelial cell biomarker SE-1.Lots of hepatic sinusoidal endothelial cells positive for SE-1 staining could be detected in CS1 group at 72 h after transplantation,while much less SE-1 positive cells presented in the control goup.Prolonged cold ischemia caused a significant increase of TNF-α,IL-1β and VEGF mRNA expression in the graft liver of control group after transplantation.The expression of TNF-α mRNA at 6 and 24 h and VEGF mRNA expression at 24 h were significantly lower in CS1 group than in control group (P＜0.05).Conclusion Peptide-mediated blockade of inflammatory cells-fibronectin interaction decreased the mRNA expression of inflammatory cytokines,prevented hepatic sinusoidal endothelial cells from injury and subsequently protected against severe ischemia/reperfusion injury of the graft liver after transplantation.