缺血预处理对大鼠肝脏缺血/再灌注早期核因子κB活性及细胞凋亡的影响  

Effects of ischemic preconditioning on cell apoptosis and the activity of nuclear factor-kappaB induced by hepatic ischemia-reperfusion injury in rats

作　　者：庄永敬[1] 曹云飞[2] 吴桂荣[1] 卢良声[1] 

机构地区：[1]解放军421医院普外科,广东广州510318 [2]解放军421医院麻醉科,广东广州510318

出　　处：《肝胆胰外科杂志》2009年第6期431-434,共4页Journal of Hepatopancreatobiliary Surgery

基　　金：国家自然科学基金资助项目(C30000158)

摘　　要：目的观察缺血预处理对大鼠肝脏缺血/再灌注早期核因子κB活性、促凋亡基因Fas和FasL蛋白表达以及肝细胞凋亡的影响,以进一步阐明肝脏缺血预处理的抗凋亡作用机制。方法建立SD大鼠肝缺血(40min)再灌注(120min)损伤及肝缺血预处理的模型。24只大鼠随机分成3组(n=8)。①假手术对照组(C组),仅分离肝十二指肠韧带,不阻断肝门,不进行其他干预处理。②缺血再灌注组(IR组),在第一肝门用小血管夹阻断尾状叶及左肝叶血流40min松开血管夹肝脏再灌注2h,再灌注开始后关腹。③缺血预处理组(IP组),先行3个循环的缺血预处理,阻断第一肝门10min,开放再灌注10min为1个循环,随后操作同缺血/再灌注组。实验结束后全自动生化分析仪检测血清谷草转氨酶和血清谷丙转氨酶活性;TUNEL法检测肝组织的细胞凋亡指数(AI);EMSA法测定肝组织核因子κB的结合活性;Westernblotting免疫印迹法检测Fas及FasL蛋白的表达水平。结果IR组和IP组的ALT、AST及细胞凋亡指数(AI)均明显高于S组(P<0.01),但与IR相比,IP组则明显较低(P<0.01)。与C组比较,IR组的核因子κB活性、促凋亡基因Fas和FasL蛋白表达水平明显增高,而IP组仅轻度增高。结论缺血预处理可通过降低肝脏缺血/再灌注早期核因子κB的转录活性,并下调促凋亡基因Fas及FasL的表达,从而发挥抗细胞凋亡和损伤保护效应。Objective To explore the effects of ischemic preconditioning on cell apoptosis, activity of NF-κB, Fas and Fas-ligand expression induced by hepatic ischemia-reperfusion injury in rats, and to elucidate the underlying anti-apoptosis mechanism of ischemic preconditioning. Methods Experimental models of hepatic ischemia-reperfusion injury were established in adult healthy SD rats, twenty-four rats were randomized into three groups （n=8）. ①Sham operation group, only hepatoduodenal ligament was separated, porta hepatis did not be blocked up, without carrying out other interventional treatments. ②IR group： the blood of caudate lobe and left lobes of liver were blocked up for 40 minutes with small vessel clip at the first porta hepatis, then small vessel clip was taken away to recover liver reperfusion for 2 hours, at the same time, rat abdomen was closed. ③IP group： the first porta hepatis was blocked up for 10 minutes followed by 10 minutes’ opening, referred to as a cycle, which was repeated for 3 times, sequent treatment was identical with I/R group. The level of aspartate aminotransferase （AST） and alanine aminotransferase （ALT） in the serum of rats were detected with automatic biochemistry analyzer, the apoptosis index （AI） of hepatic cells was measured with TUNEL method, DNA binding activity of NF-κB in liver grafts was analyzed with EMSA （electrophoretic mobility shift assay）, and the Fas and Fas ligand expression in liver tissue was assessed with immunohistochemical method. Results The levels of ALT, AST and AI were significantly higher in IR group and IP group than that in S group （P 〈0.01）, and remarkable higher were also observed in IR group than those in IP group （P〈0.01）. Compared with C group, activity of NF-κB, Fas and Fas-ligand expression were significantly increased in IR group, while only modest increase was observed in IP group. Conclusion Ischemic preconditioning can relieve the liver cellular apoptosis by regulating the activity of NF-κB, Fas and Fas

关 键 词：缺血预处理 肝脏 再灌注损伤 核因子ΚB 凋亡 大鼠 

分 类 号：R657.3[医药卫生—外科学]