肝窦内皮细胞HO-1表达促进小鼠肝脏缺血再灌注损伤后再生重建的机制研究  

Mechanism of HO-1 expression in hepatic sinusoidal endothelial cells promoting liver regeneration and reconstruction after ischemia-reperfusion injury in mice

作　　者：李鹏程 王路兵 朱荣华 龚庆豪 LI Peng-cheng;WANG Lu-bing;ZHU Rong-hua;GONG Qing-hao(Department of General Surgery,Chongming Hospital,Shanghai Health Medical College,Shanghai,202150,China)

机构地区：[1]上海健康医学院附属崇明医院普外科,202150

出　　处：《肝脏》2025年第1期122-127,共6页Chinese Hepatology

基　　金：上海市崇明区科技项目(CKY2022-05)。

摘　　要：目的探讨肝窦内皮细胞(LSEC)血红素加氧酶-1(HO-1)表达促进小鼠肝脏缺血再灌注损伤后再生重建的机制。方法60只小鼠分为假手术组、模型组、空载组、HO-1转染组,每组15只,除假手术组仅做麻醉开腹操作外,其余3组均使用经典肝脏70%热缺血方法构建小鼠肝脏缺血再灌注损伤模型,再灌注后空载组经门静脉注射200μL空载体病毒转染的LSEC,HO-1转染组经门静脉注射200μL CD31标记的2×10^(6)个/mL HO-1转染的LSEC。比较各组小鼠肝功能、血清及肝组织炎症因子、氧化应激因子水平、肝组织内肝细胞生长因子(HGF)、凋亡蛋白、Nrf2/HO-1信号通路蛋白表达量变化及肝细胞凋亡情况。结果携带HO-1目的基因的慢病毒的HO-1 mRNA表达量显著高于空载组(P<0.05)。模型组、空载组和HO-1转染组的ALT和AST水平高于假手术组,HO-1转染组的ALT和AST水平低于模型组和空载组(P<0.05);模型组和空载组比较差异无统计学意义(P>0.05)。模型组、空载组和HO-1转染组血清及肝组织的TNF-α、IL-1β、IL-6和MDA水平高于假手术组,SOD水平降低(P<0.05);HO-1转染组血清及肝组织的TNF-α、IL-1β、IL-6和MDA水平低于模型组和空载组,SOD升高(P<0.05);模型组和空载组比较差异无统计学意义(P>0.05)。模型组、空载组和HO-1转染组肝细胞病理变化程度和凋亡数高于假手术组,HO-1转染组低于模型组和空载组(P<0.05);模型组和空载组比较差异无统计学意义(P>0.05)。模型组、空载组和HO-1转染组肝组织的Bax和Caspase-3水平高于假手术组,HGF和Bcl-2降低(P<0.05);HO-1转染组肝组织的Bax和Caspase-3水平低于模型组和空载组,HGF和Bcl-2升高(P<0.05);模型组和空载组比较差异无统计学意义(P>0.05)。模型组、空载组和HO-1转染组肝组织的Nrf2和HO-1水平低于假手术组(P<0.05),HO-1转染组肝组织的Nrf2和HO-1水平高于于模型组和空载组(P<0.05);模型组和空载组比较差异无统计学意Objective To investigate the mechanism of heme oxygenase-1(HO-1)expression in hepatic sinusoidal endothelial cells promoting liver regeneration and reconstruction after ischemia-reperfusion injury in mice.Methods Sixty male mice were randomly divided into a sham surgery group,a model group,an empty vector group,and a HO-1 transfection group of 15 mice each.Except for the sham surgery group,which only underwent anesthesia and open surgery,the other three groups used the classic liver 70%warm ischemia method to establish a mouse model of liver ischemia-reperfusion injury.After reperfusion,the empty vector group was injected with 200μL of empty vector virus transfected LSECs via the portal vein,while the HO-1 transfection group was injected with 200μL of CD31 labeled 2×10^(6)/mL HO-1 transfected LSECs via the portal vein.Evaluate liver function indicators[alanine aminotransferase(ALT),aspartate aminotransferase(AST)],serum and liver tissue inflammatory factors[tumor necrosis factor]among different groups of mice-α(TNF-α),Interleukin-1β(IL-1β),changes in levels of interleukin-6(IL-6),oxidative stress factors superoxide dismutase(SOD),malondialdehyde(MDA),hepatocyte growth factor(HGF),apoptotic proteins Bax,Bcl-2,Caspase-3,and nuclear factor E2 related factor 2(Nrf2)and HO-1 protein expression in the Nrf2/HO-1 signaling pathway were observed in liver tissue.Pathological changes in liver tissue and liver cell apoptosis rate were also observed.Results RT-PCR assay showed that the mRNA expression of HO-1 in lentivirus carrying HO-1 target gene was significantly higher than that in no-load group(P<0.05).ALT and AST levels in model group,no-load group and HO-1 transfection group were higher than those in sham operation group,while ALT and AST levels in HO-1 transfection group were lower than those in model group and no-load group(P<0.05).There was no difference between model group and no-load group(P>0.05).The levels of TNF-α,IL-1β,IL-6 and MDA in serum and liver tissue of model group,no-load group,and HO-1 transf

关 键 词：肝脏缺血再灌注损伤 肝窦内皮细胞 血红素加氧酶-1 再生重建 

分 类 号：R657.3[医药卫生—外科学]