冷保存供肝肝移植大鼠肝窦内皮细胞损伤机制的研究

目的 探寻肝移植大鼠肝窦内皮细胞(SEC)损伤的详细过程、方式及机制,为冷保存再灌注损伤的保护研究开辟新的途径.方法 雄性SD大鼠随机分为假手术组(n=6)、UW 1 h肝移植组(n:48)、Uw 12 h肝移植组(n=48).大鼠原位肝移植采用双袖套法,分别于术后不同时相点采取血液及组织标本,检测血清丙氨酸氨基转移酶(ALT)及透明质酸(HA)水平;HE染色观察肝脏病理学变化;TUNEL法检测凋亡,免疫组化法检测Bcl-2及Cleaved Caspase-3的表达状况.结果 UW 1 h、UW 12 h组肝移植后血清ALT、HA均较假手术组明显升高(P<0.05),UW 12 h组又明显高于Uw 1 h组(P<0.05).UW 12 h组ALT水平于术后6 h达高峰,而HA水平却在术后1 h、24 h呈双峰表现.Uw 12 h组首先出现SEC的凋亡继而出现肝细胞的坏死,且UW 12 h组细胞凋亡指数(apoptosis index,AI)明显高于UW 1 h组(P<0.01).两组大鼠SEC的AI均于术后6 h达高峰,与血中ALT的高峰时相点一致.肝移植术后Bcl-2表达明显减弱(P

Mechanisms of injury to sinusoidal endothelial cells after extended preservation and transplantation of rat liver

Objective To investigate the detailed process and mechanisms of injury to sinusoidal endothelial cells(SECs)after cold-preserved transplantation of rat liver. Methods Male rats were randomized in-to three groups: sham-operated group,UW 1 h group and UW 12 h group. Orthotopic liver transplantation (OLT)was performed using the two-cuff technique devised by Kamada with some modifications. Liver tissuespecimens and blood samples were collected at different time points after operation. The liver function was evaluated by serum ALT and HA levels.Morphology was observed with light microscopy. The incidence of apoptosis was deternmined by TUNEL method. Additionally, the expression of Bcl-2 and Cleaved Caspase-3 was examined by immunohistochemistry. Results The levels of serum ALT and HA were significantly higher in UW 12 h group than those in UW 1 h group(P