大鼠脂肪干细胞对自体肝移植缺血再灌注损伤的保护机制研究  

Protective effects of adipose-derived stem cells on ischemia-reperfusion injury after liver autotransplantation in rats

作　　者：倪孝龙 何其宽 赵炜嘉 余华军[1] 鲍海力 张启瑜[1] NI Xiao-long;HE Qi-kuan;ZHAO Wei-jia;YU Hua-jun;BAO Hai-li;ZHANG Qi-yu(Department of Hepatobiliary Surgery, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou,Zhejiang 325000, China)

机构地区：[1]温州医科大学附属第一医院肝胆外科,浙江温州325000

出　　处：《肝胆胰外科杂志》2017年第2期128-133,共6页Journal of Hepatopancreatobiliary Surgery

基　　金：国家自然科学基金项目(81470874)

摘　　要：目的探讨大鼠脂肪干细胞在大鼠自体肝移植缺血再灌注(ischemia-reperfusion,IR)损伤模型中的保护作用机制。方法分离并培养大鼠脂肪干细胞(adipose-derived stem cells,ADSCs),构建稳定细胞株。SD大鼠随机分为假手术组、自体肝移植缺血再灌注组(IR组)、自体肝移植缺血再灌注+脂肪干细胞回填组(ADSCs组),每组10只。假手术组上腹正中切口,暴露并离断肝脏韧带,不做其他处理。IR组用动脉夹夹闭肝门阻断血流,肝素注射液灌注15 min,不给药。ADSCs组在肝素注射液灌注后30 min尾静脉注射1×106 ADSCs。手术后24 h后处死大鼠,检测各组大鼠血清中天冬氨酸氨基转移酶(AST)、丙氨酸转氨酶(ALT)、肝脏超氧化物歧化酶(SOD)、丙二醛(MDA)含量,以及线粒体活性;苏木素-伊红染色(HE)观察肝脏病理学改变;免疫印迹法(Western blotting)检测细胞间黏附分子-1(ICAM-1)、Bax基因表达情况。结果与假手术组相比,IR组AST和ALT水平、MDA含量较高,SOD含量较低(P<0.05),线粒体活性较弱。病理切片结果显示,IR组肝细胞出现坏死,炎性细胞浸润严重。Western blotting结果显示IR组较假手术组ICAM-1、Bax表达增多。与IR组相比,ADSCs组AST和ALT水平、MDA含量较低,SOD含量较高(P<0.05),线粒体活性较强,炎性细胞浸润较少,ICAM-1、Bax表达减少。结论大鼠脂肪干细胞对大鼠自体肝移植缺血再灌注损伤模型有保护作用,其作用机制可能是通过减少炎症反应和氧化应激程度进而减轻肝脏缺血再灌注损伤。Objective To investigate the protective mechanism of adipose-derived stem cells(ADSCs)onischemia-reperfusion(IR)injury after liver auto-transplantation in rats.Methods Rat ADSCs were isolatedand purified from adipose tissues of rat.Adult male SD rats were randomly divided into Sham-operated normalcontrol group(n=10),ischemia-reperfusion injury group(IR group,n=10)and ischemia-reperfusion injury withintravenous adipose-derived stem cells group(ADSCs group,n=10).In the sham-operated group,the medianincision of the upper abdomen was performed,and ligament of the liver was exposed and disconnected withoutany other treatment.In IR group,the portal vein was clamped by arterial clamp to block the blood flow;heparininjection was perfused for15min without any administration.ADSCs group was injected with1×106ADSCsin the tail vein30min after heparin infusion.All rats were sacrificed24h after reperfusion.Serum aspartatetransaminase(AST),alanine aminotransferase(ALT),superoxide dismutase(SOD),malondialdehyde(MDA),vitality of mitochondria were detected.The pathological changes of liver tissue were assessed with haematoxylinand eosin-stained(HE).The expression of intercellular adhesion molecule(ICAM-1),Bax were detected withWestern blotting method.Results Compared with the sham-operated group,the levels of AST,ALT and MDAincreased;levels of SOD and vitality of mitochondria decreased in IR group(P<0.05).Pathological results suggestedthat IR group showed necrosis of hepatocytes and severe inflammatory cell infiltration.Western blotting results suggested that,compared with the sham-operated group,expression of ICAM-1and Bax increased in IRgroup.Compared with the IR group,levels of AST,ALT and MDA decreased,levels of SOD and vitality of mitochondriaincreased(P<0.05),neutrophil infiltration into the liver was fewer in ADSCs group.Compared with IRgroup,the expression of ICAM-1and Bax decreased in ADSCs group.Conclusion The results indicate that ADSCsmay protect liver from ischemia-reperfusion injury in rats model.The mechanism invol

关 键 词：脂肪干细胞 缺血再灌注损伤 自体肝移植 大鼠 

分 类 号：R657.3[医药卫生—外科学]