摘要
目的观察盲肠结扎穿孔术(CLP)诱导的脓毒症小鼠肝损伤时不同途径铁死亡的发生,探讨线粒体乙醛脱氢酶2(ALDH2)是否通过抑制铁死亡减轻脓毒症肝损伤。方法选取60只8周龄雄性C57BL/6J小鼠,按随机数字表法分为假手术(Sham)组、CLP组、铁死亡抑制剂铁抑素-1(Fer-1)组、ALDH2特异性激动剂Alda-1组、铁螯合剂右雷佐生(DXZ)组、二甲基亚砜(DMSO)组,每组10只。采用CLP法建立小鼠脓毒症肝损伤模型;Sham组仅开腹不结扎和穿刺盲肠。DMSO组、Fer-1组、DXZ组和Alda-1组分别于CLP术前1 h腹腔注射10 mL/kg 5%DMSO、5 mg/kg Fer-1、50 mg/kg DXZ和10 mg/kg Alda-1。术后24 h,麻醉小鼠取眼球血及肝组织。苏木素-伊红(HE)染色,镜下观察肝脏结构和炎症浸润情况;检测各组血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)水平及肝组织丙二醛(MDA)、超氧化物歧化酶(SOD)、活性氧(ROS)水平;蛋白质免疫印迹试验(Western blotting)检测肝组织ALDH2、铁死亡相关蛋白谷胱甘肽过氧化物酶4(GPX4)、铁死亡抑制蛋白1(FSP1)和转铁蛋白受体1(TFR1)的蛋白表达。结果与Sham组相比,CLP组小鼠术后24 h肝脏可见不同程度淤血、肝细胞排列紊乱及炎症细胞浸润;与CLP组相比,Fer-1组、DXZ组和Alda-1组小鼠肝脏形态学有改善,损伤减轻,炎症细胞浸润减少。与Sham组相比,CLP组血清ALT、AST水平及肝组织MDA、ROS含量和TFR1蛋白表达显著升高,肝组织SOD活性及ALDH2、GPX4和FSP1蛋白表达显著降低。与CLP组相比,Fer-1组、DXZ组和Alda-1组血清ALT、AST水平显著降低〔ALT(U/L):45.76±10.81、37.30±2.98、36.40±12.75比73.06±12.20,AST(U/L):61.57±2.69、52.41±6.92、56.05±8.29比81.59±5.46,均P<0.05〕,肝组织MDA、ROS含量及TFR1蛋白表达显著降低〔MDA(μmol/L):0.60±0.10、0.57±0.18、0.83±0.39比1.61±0.30,ROS(荧光强度):270.34±9.64、276.02±62.33、262.05±18.55比455.38±36.07,TFR1/GAPDH:0.90±0.04、1.01±0.09、0.55±0.08比1.18±0.0
Objective To observe the ferroptosis triggered by in different pathways during cecal ligation and puncture(CLP)-induced liver injury in septic mice,and to investigate whether mitochondrial aldehyde dehydrogenase 2(ALDH2)can alleviate sepsis-induced liver injury by inhibiting ferroptosis.Methods Sixty 8-week-old male C57BL/6J mice were randomly divided into sham operation group(Sham group),CLP group,ferroptosis inhibitor ferrostain-1(Fer-1)group,ALDH2-specific agonist Alda-1 group,iron chelator deferasirox Fe3+chelate(DXZ)group and dimethyl sulfoxide(DMSO)group,with 10 mice in each group.The septic liver injury was induced by CLP in mice model.In the Sham group,only laparotomy was performed without ligation and puncture of the cecum.10 mL/kg 5%DMSO,5 mg/kg Fer-1,50 mg/kg DXZ and 10 mg/kg Alda-1 were injected intraperitoneally 1 hour before CLP in the DMSO,Fer-1,DXZ and Alda-1 groups respectively.At 24 hours after operation,eyeball blood and liver tissue were collected from anesthetized mice.The hepatic structure and inflammatory infiltration were observed by hematoxylin-eosin(HE)staining.The levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST)in serum,the levels of hepatic malondialdehyde(MDA),superoxide dismutase(SOD)and reactive oxygen species(ROS)were detected.Western blotting was used to detect the protein expressions of ALDH2,ferroptosis-related proteins glutathione peroxidase 4(GPX4),ferroptosis suppressor protein 1(FSP1)and transferrin receptor 1(TFR1)in liver tissue.Results Compared with Sham group,the mice in CLP group showed varying degrees of congestion,disorganized hepatocyte arrangement,inflammatory cell infiltration at 24 hours after operation.Compared with the CLP group,the mice in the Fer-1 group,DXZ group and Alda-1 group liver morphology,liver injury and inflammatory cell infiltration was improved.Compared with Sham group,the serum levels of ALT and AST,the contents of MDA and ROS,and the expression of TFR1 protein in CLP group were significantly increased,while the activity of
作者
李璐
李玉萍
陆娜
杜若丽
李文韬
叶红伟
高琴
Li Lu;Li Yuping;Lu Na;Du Ruoli;Li Wentao;Ye Hongwei;Gao Qin(Department of Physiology,Key Laboratory of Basic and Clinical Cardiovascular Diseases,Bengbu Medical College,Bengbu 233000,Anhui,China;Department of Respiratory and Critical Care Medicine,the First Affiliated Hospital of Bengbu Medical College,Bengbu 233000,Anhui,China;Clinical Medical College,Bengbu Medical College,Bengbu 233000,Anhui,China)
出处
《中华危重病急救医学》
CAS
CSCD
北大核心
2023年第7期684-689,共6页
Chinese Critical Care Medicine
基金
国家自然科学基金(81770297)。
关键词
线粒体乙醛脱氢酶2
脓毒症肝损伤
铁死亡
Mitochondrial acetaldehyde dehydrogenase 2
Septic liver injury
Ferroptosis
