Nrf2激动剂萝卜硫素通过抑制肝组织铁死亡减轻小鼠脓毒症肝损伤  

Nrf2 agonist sulforaphane attenuates liver injury in septic mice by inhibiting ferroptosis

作　　者：魏嘉亿 张钦 李乐谦 黄小夏 刘转华 孙毛毛 包俊颖 陈红宇 郭晓华 黄巧冰 WEI Jia-yi;ZHANG Qin;LI Le-qian;HUANG Xiao-xia;LIU Zhuan-hua;SUN Mao-mao;BAO Jun-ying;CHEN Hong-yu;GUO Xiao-hua;HUANG Qiao-bing(Department of Pathophysiology,School of Basic Medical Sciences,Guangdong Provincial Key Lab of Shock and Microcir-culation,Southern Medical University,Guangzhou 510515,China;The First School of Clinical Medicine,Southern Medi-cal University,Guangzhou 510515,China)

机构地区：[1]南方医科大学基础医学院病理生理学教研室,广东省医学休克微循环重点实验室,广东广州510515 [2]南方医科大学第一临床医学院,广东广州510515

出　　处：《中国病理生理杂志》2022年第4期608-615,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金面上项目(No.82172139)。

摘　　要：目的:探讨核因子E2相关因子2(nuclear factor E2-related factor 2,Nrf2)激动剂萝卜硫素(sulfora⁃phane,SFN)对脓毒症小鼠肝脏铁死亡的影响及其分子机制。方法:用盲肠结扎穿孔术(cecal ligation and punc⁃ture,CLP)复制C57BL/6小鼠的脓毒症模型。将动物随机分成4组:假手术(sham)组、CLP组、CLP+SFN组和CLP+去铁胺(DFO)组,每组4只。检测CLP术后12 h各组小鼠肝组织非血红素铁、白细胞介素6(interleukin-6,IL-6)mRNA、HAMP(hepcidin antimicrobial peptide;编码铁调素)mRNA、前列腺素内过氧化物合成酶2(prostaglandin-en⁃doperoxide synthase 2,PTGS2)mRNA和铁输出蛋白1(ferroportin 1,FPN1)水平,以及血清IL-6、铁调素等铁代谢和铁死亡相关指标;检测血清丙氨酸转氨酶(alanine aminotransferase,ALT)和天冬氨酸转氨酶(aspartate aminotrans⁃ferase,AST)活性,以及肝组织脂质过氧化水平如还原型烟酰胺腺嘌呤二核苷酸磷酸(reduced nicotinamide adenine dinucleotide phosphate,NADPH)和丙二醛(malondialdehyde,MDA)的变化,并观察肝组织形态学改变;培养巨噬细胞证实SFN对Nrf2活性的影响。结果:与sham组相比,CLP组12 h可见肝组织炎性浸润,血清中ALT和AST活性升高;肝组织非血红素铁含量增高,HAMP mRNA增多,FPN1蛋白量降低;电镜可见线粒体缩小和嵴消失,并伴随血清IL-6含量和铁调素含量的增加,证明CLP可导致肝组织铁超载和铁死亡。在证实SFN可恢复被脂多糖(lipopoly⁃saccharide,LPS)下调的巨噬细胞Nrf2表达的基础上,证明给予SFN可明显降低CLP组小鼠血清IL-6含量,减轻肝组织铁超载,缓解上述CLP小鼠的铁代谢障碍,恢复肝细胞线粒体结构,减轻铁死亡;并减少肝组织炎细胞浸润,降低血清ALT和AST活性。结论:Nrf2激活剂SFN可减轻CLP引起的小鼠肝组织铁死亡,保护肝功能;其作用可能与调节CLP小鼠肝组织的IL-6/铁调素/FPN1通路活性有关。AIM:To investigate the effect of nuclear factor E2-related factor 2(Nrf2)agonist sulforaphane(SFN)on liver ferroptosis in septic mice and its mechanism.METHODS:The sepsis model of C57BL/6 mice was in⁃duced by cecal ligation and puncture(CLP)method.The animals were randomly divided into 4 groups:sham group,CLP group,CLP+SFN group and CLP+deferroamine(DFO)group,with 4 mice in each group.Twelve hours after CLP induc⁃tion,the markers of iron metabolism and ferroptosis,including non-heme iron,interleukin-6(IL-6)mRNA,hepcidin an⁃timicrobial peptide(HAMP;encoding hepcidin)mRNA,prostaglandin-endoperoxide synthase 2(PTGS2)mRNA and fer⁃roportin 1(FPN1)in liver tissue,as well as serum IL-6 and hepcidin,were detected.The levels of alanine aminotransfer⁃ase(ALT)and aspartate aminotransferase(AST)in serum,and reduced nicotinamide adenine dinucleotide phosphate(NADPH)and malondialdehyde(MDA)in liver tissue were measured.The morphological changes of liver tissue were ob⁃served.The effect of SFN on Nrf2 expression was confirmed in cultured macrophages.RESULTS:Compared with sham group,the levels of ALT and AST in serum were increased and inflammatory cell infiltration was presented in liver tissue at 12 h in CLP group.The levels of non-heme iron and HAMP mRNA were also increased,while FPN1 protein expression was decreased,accompanied by increases in serum IL-6 and hepcidin levels in CLP group.There were mitochondrial shrinkage and cristae disappearance in liver cells.Treatment with SFN restored lipopolysaccharide(LPS)-attenuated ex⁃pression of Nrf2 in macrophages,significantly reduced serum IL-6 content in CLP mice,and alleviated iron overload in liv⁃er tissue and iron metabolism disorder in CLP mice.The mitochondrial structure of liver cells was also restored.The infil⁃tration of inflammatory cells in liver tissue and the ALT and AST levels in serum were decreased in CLP+SFN group.CONCLUSION:The Nrf2 agonist SFN can inhibit CLP-induced ferroptosis in liver tissue and protect liver function in mice.These effects

关 键 词：核因子E2相关因子2 萝卜硫素 脓毒症 肝损伤 铁死亡 

分 类 号：R631[医药卫生—外科学] R363.1[医药卫生—临床医学]