基于铁死亡探讨乌司他丁对肝缺血-再灌注损伤的保护作用  

Protective effect of ulinastatin on hepatic ischemia-reperfusion injury based on ferroptosis

作　　者：陈实 赵漾 周尧 郁冬玲 黄娇 蓝雨雁[1] Chen Shi;Zhao Yang;Zhou Yao;Yu Dongling;Huang Jiao;Lan Yuyan(Department of Anesthesiology,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China;不详)

机构地区：[1]广西医科大学第一附属医院麻醉科,南宁530021 [2]广西医学科学院广西壮族自治区人民医院急诊科

出　　处：《器官移植》2024年第5期780-788,共9页Organ Transplantation

基　　金：广西自然科学基金(2018GXNSFAA281101)。

摘　　要：目的探讨乌司他丁对肝缺血-再灌注损伤的保护作用及机制。方法24只雄性SD大鼠分成3组,分别为假手术组(Sham组)、肝缺血-再灌注损伤组(HIRI组)、肝缺血-再灌注损伤+乌司他丁预处理组(HIRI+UTI组),每组8只。采用阻断肝门静脉和肝动脉1 h的方法建立大鼠HIRI模型,HIRI+UTI组于造模前30 min腹腔注射乌司他丁,Sham组和HIRI组腹腔注射等量的生理盐水。造模6 h后处死大鼠,收集大鼠血清检测丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)水平;采用苏木素-伊红(HE)染色法检测肝组织病理学改变;透射电镜观察肝组织线粒体超微结构改变;免疫荧光检测谷胱甘肽过氧化酶4(GPX4)表达;检测肝组织丙二醛(MDA)、谷胱甘肽(GSH)、铁、活性氧簇(ROS)及GPX4含量;检测肝组织GPX4、酰基辅酶A合成酶长链家族4(ACSL4)信使RNA(mRNA)和蛋白表达水平。结果与Sham组相比,HIRI组血清ALT、AST水平升高,肝脏出现淤血、肝细胞坏死和肝小叶结构异常等病理改变,病理学评分升高,线粒体缩小,膜密度增加,线粒体嵴断裂甚至消失,ROS、MDA、铁含量升高,GSH含量下降,GPX4荧光强度减弱,ACSL4 mRNA及蛋白相对表达量升高,GPX4 mRNA和蛋白相对表达量降低(均为P<0.05)。与HIRI组比较,HIRI+UTI组血清ALT、AST水平降低,肝组织损伤减轻,病理学评分降低,线粒体缩小、嵴断裂情况改善,ROS、MDA、铁含量降低,GSH含量升高,GPX4荧光强度增强,ACSL4 mRNA和蛋白相对表达量降低,GPX4 mRNA和蛋白相对表达量升高(均为P<0.05)。结论乌司他丁可减轻大鼠肝缺血-再灌注损伤,其机制可能是通过抑制铁死亡。Objective To evaluate the protective effect and underlying mechanism of ulinastatin on hepatic ischemia-reperfusion injury.Methods Twenty-four male SD rats were divided into three groups:sham operation group(Sham group),hepatic ischemia-reperfusion injury group(HIRI group)and hepatic ischemia-reperfusion injury+ulinastatin pretreatment group(HIRI+UTI group),with 8 rats in each group.The HIRI rat models were established by occluding hepatic portal vein and hepatic artery for 1 h.In the HIRI+UTI group,the rats were intraperitoneally injected with ulinastatin at 30 min before model establishment,and an equivalent amount of normal saline was given in the Sham and HIRI groups.The rats were sacrificed at 6 h after model establishment.Serum samples were collected to detect alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels.Pathological changes of liver tissues were observed by hematoxylin-eosin(HE)staining.Ultrastructural changes of mitochondria in liver tissues were observed by transmission electron microscopy.The expression of glutathione peroxidase 4(GPX4)was determined by immunofluorescent staining.The contents of malondialdehyde(MDA),glutathione(GSH),Fe,reactive oxygen species(ROS)and GPX4 were detected.The expression levels of GPX4 and acyl-CoA synthetase long-chain family 4(ACSL4)messenger RNA(mRNA)and proteins in liver tissue were measured.Results Compared with the Sham group,serum ALT and AST levels were up-regulated,pathological changes such as congestion,hepatocyte necrosis and abnormal hepatic lobule structure were observed,pathological score was increased,the mitochondria shrank,the membrane density was increased,the mitochondrial crest was damaged or even absent,the contents of ROS,MDA and Fe were elevated,the GSH content was decreased,the fluorescent intensity of GPX4 was weakened,the relative expression levels of ACSL4 mRNA and protein were up-regulated,and the relative expression levels of GPX4 mRNA and protein were down-regulated in the HIRI group(all P<0.05).Compared with the HIRI

关 键 词：缺血-再灌注损伤 肝移植 乌司他丁 铁死亡 谷胱甘肽过氧化酶4 酰基辅酶A合成酶长链家族4 活性氧簇 丙二醛 

分 类 号：R617[医药卫生—外科学] R657.3[医药卫生—临床医学]