铁死亡在细菌性感染中的研究进展

doi:10.11843/j.issn.0366-6964.2023.06.008

畜牧兽医学报 ›› 2023, Vol. 54 ›› Issue (6): 2280-2287.doi: 10.11843/j.issn.0366-6964.2023.06.008

铁死亡在细菌性感染中的研究进展

毛鹏^1,2,3, 王志浩^1,2,3, 李建基^1,2,3, 崔璐莹^1,2,3, 朱国强^1,2,3, 孟霞^1,2,3, 董俊升^1,2,3, 王亨^1,2,3*

1. 扬州大学兽医学院江苏省动物重要疫病与人兽共患病防控协同创新中心, 扬州 225009;
2. 江苏高校动物重要疫病和重要人兽共患病防控技术国际合作联合实验室, 扬州 225009;
3. 农业与农产品安全教育部国际联合研究实验室, 扬州 225009

收稿日期:2022-07-21 出版日期:2023-06-23 发布日期:2023-06-16
通讯作者: 王亨,主要从事动物临床疾病诊疗和发病机制研究,E-mail:sdaulellow@163.com
作者简介:毛鹏(1996-),男,甘肃酒泉人,博士生,主要从事动物临床疾病研究,E-mail:maopeng96@163.com
基金资助:
国家自然科学基金(32273070);江苏省自然科学基金面上项目(BK20211324);江苏现代农业产业技术体系建设专项资金(JATS[2022]499);江苏省333高层次人才培养工程资助项目;高等学校学科创新引智计划资助(D18007);江苏高校优势学科建设工程资助项目;江苏高校动物重要疫病和重要人兽共患病防控技术国际合作联合实验室资助;江苏高校品牌专业建设工程资助项目

Research Progress of Ferroptosis in Bacterial Infection

MAO Peng^1,2,3, WANG Zhihao^1,2,3, LI Jianji^1,2,3, CUI Luying^1,2,3, ZHU Guoqiang^1,2,3, MENG Xia^1,2,3, DONG Junsheng^1,2,3, WANG Heng^1,2,3*

1. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China;
2. International Research Laboratory of Prevention and Control of Important Animal Infectious Diseases and Zoonotic Diseases of Jiangsu Higher Education Institutions, Yangzhou 225009, China;
3. Joint International Research Laboratory of Agriculture and Agri-Product Safety, the Ministry of Education, Yangzhou 225009, China

Received:2022-07-21 Online:2023-06-23 Published:2023-06-16

摘要/Abstract

摘要： 铁死亡是一种新型程序性细胞死亡模式,其主要特点为铁依赖的脂质过氧化物蓄积。在细菌感染宿主细胞的过程中,铁代谢紊乱和氧化应激起到了重要作用,并且已经证明某些细菌可通过铁死亡的方式诱导宿主细胞死亡。本文就细菌感染对宿主铁代谢的影响以及细菌诱导铁死亡的研究现状进行阐述,以期为防控动物细菌感染提供新的思路。

关键词: 铁死亡, 细菌感染, 铁代谢, 氧化应激

Abstract: Ferroptosis is a newly discovered programmed cell death pattern characterized by iron-dependent accumulation of lipid peroxidation. Iron metabolism disorder and oxidative stress play an important role in bacterial infection, and it has been proved that some bacteria could induce host cell death by ferroptosis. In this paper, the effects of bacterial infection on host iron metabolism and the mechanism of bacteria-induced ferroptosis were reviewed, in order to provide new research ideas for the prevention of bacterial infection in animals.

Key words: ferroptosis, bacterial infection, iron metabolism, oxidative stress

中图分类号:

S855.1

毛鹏, 王志浩, 李建基, 崔璐莹, 朱国强, 孟霞, 董俊升, 王亨. 铁死亡在细菌性感染中的研究进展[J]. 畜牧兽医学报, 2023, 54(6): 2280-2287.

MAO Peng, WANG Zhihao, LI Jianji, CUI Luying, ZHU Guoqiang, MENG Xia, DONG Junsheng, WANG Heng. Research Progress of Ferroptosis in Bacterial Infection[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2280-2287.

参考文献 70

[1]	DIXON S J,LEMBERG K M,LAMPRECHT M R,et al.Ferroptosis:An iron-dependent form of nonapoptotic cell death[J].Cell,2012,149(5):1060-1072.
[2]	LIN R Y,ZHANG Z H,CHEN L F,et al.Dihydroartemisinin (DHA) induces ferroptosis and causes cell cycle arrest in head and neck carcinoma cells[J].Cancer Lett,2016,381(1):165-175.
[3]	MA S,HENSON E S,CHEN Y,et al.Ferroptosis is induced following siramesine and lapatinib treatment of breast cancer cells[J].Cell Death Dis,2016,7(7):e2307.
[4]	DO VAN B,GOUEL F,JONNEAUX A,et al.Ferroptosis,a newly characterized form of cell death in Parkinson's disease that is regulated by PKC[J].Neurobiol Dis,2016,94:169-178.
[5]	GAO M H,MONIAN P,QUADRI N,et al.Glutaminolysis and transferrin regulate ferroptosis[J].Mol Cell,2015,59(2):298-308.
[6]	陈敬宜,于淼,张金洋,等.铁死亡参与镉暴露鸡肝损伤的研究[J].畜牧兽医学报,2023,54(2):787-802.CHEN J Y,YU M,ZHANG J Y,et al.Study on the involvement of ferroptosis in liver injury of cadmium-exposed chickens[J].Acta Veterinaria et Zootechnica Sinica,2023,54(2):787-802.(in Chinese)
[7]	刘武康,吴淑燕,陈国薇,等.细菌产生的活性氧及其功能[J].微生物学杂志,2016,36(1):89-95.LIU W K,WU S Y,CHEN G W,et al.The reactive oxygen species generated by bacteria and its functions[J].Journal of Microbiology,2016,36(1):89-95.(in Chinese)
[8]	DOLMA S,LESSNICK S L,HAHN W C,et al.Identification of genotype-selective antitumor agents using synthetic lethal chemical screening in engineered human tumor cells[J].Cancer Cell,2003,3(3):285-296.
[9]	WANG L Y,LIU Y C,DU T T,et al.ATF3 promotes erastin-induced ferroptosis by suppressing system Xc-[J].Cell Death Differ,2020,27(2):662-675.
[10]	DIXON S J,PATEL D N,WELSCH M,et al.Pharmacological inhibition of cystine-glutamate exchange induces endoplasmic reticulum stress and ferroptosis[J].Elife,2014,3:e2523.
[11]	XIE Y C,ZHU S,SONG X X,et al.The tumor suppressor p53 limits ferroptosis by blocking DPP4 activity[J].Cell Rep,2017,20(7):1692-1704.
[12]	CHEN Y,LIU Y,LAN T,et al.Quantitative profiling of protein carbonylations in ferroptosis by an aniline-derived probe[J].J Am Chem Soc,2018,140(13):4712-4720.
[13]	BERSUKER K,HENDRICKS J M,LI Z P,et al.The CoQ oxidoreductase FSP1 acts parallel to GPX4 to inhibit ferroptosis[J].Nature,2019,575(7784):688-692.
[14]	PITMAN K E,ALLURI S R,KRISTIAN A,et al.Influx rate of18F-fluoroaminosuberic acid reflects cystine/glutamate antiporter expression in tumour xenografts[J].Eur J Nucl Med Mol Imaging,2019,46(10):2190-2198.
[15]	梅胜兰,夏中元,孟庆涛,等.细胞铁死亡发生机制的研究进展[J].医学综述,2020,26(21):4207-4211,4218.MEI S L,XIA Z Y,MENG Q T,et al.Research advances in mechanism of ferroptosis in cells[J].Medical Recapitulate,2020,26(21):4207-4211,4218.(in Chinese)
[16]	INGOLD I,BERNDT C,SCHMITT S,et al.Selenium utilization by GPX4 is required to prevent hydroperoxide-induced ferroptosis[J].Cell,2018,172(3):409-422.e21.
[17]	CAO J Y,DIXON S J.Mechanisms of ferroptosis[J].Cell Mol Life Sci,2016,73(11-12):2195-2209.
[18]	VANDEN BERGHE T,LINKERMANN A,JOUAN-LANHOUET S,et al.Regulated necrosis:the expanding network of non-apoptotic cell death pathways[J].Nat Rev Mol Cell Biol,2014,15(2):135-147.
[19]	RIEGMAN M,SAGIE L,GALED C,et al.Ferroptosis occurs through an osmotic mechanism and propagates independently of cell rupture[J].Nat Cell Biol,2020,22(9):1042-1048.
[20]	ANGELI J P F,SCHNEIDER M,PRONETH B,et al.Inactivation of the ferroptosis regulator Gpx4 triggers acute renal failure in mice[J].Nat Cell Biol,2014,16(12):1180-1191.
[21]	NISHIZAWA H,MATSUMOTO M,CHEN G,et al.Lipid peroxidation and the subsequent cell death transmitting from ferroptotic cells to neighboring cells[J].Cell Death Dis,2021,12(4):332.
[22]	KIM H J,JOE H I,ZHANG Z Y,et al.Anti-inflammatory effect of Acalypha australis L.via suppression of NF-κB signaling in LPS-stimulated RAW 264.7 macrophages and LPS-induced septic mice[J].Mol Immunol,2020,119:123-131.
[23]	LIU P F,FENG Y T,LI H W,et al.Ferrostatin-1 alleviates lipopolysaccharide-induced acute lung injury via inhibiting ferroptosis[J].Cell Mol Biol Lett,2020,25(1):10-24.
[24]	HE R Y,LIU B H,XIONG R,et al.Itaconate inhibits ferroptosis of macrophage via Nrf2 pathways against sepsis-induced acute lung injury[J].Cell Death Discov,2022,8(1):43.
[25]	LI J C,LU K M,SUN F L,et al.Panaxydol attenuates ferroptosis against LPS-induced acute lung injury in mice by Keap1-Nrf2/HO-1 pathway[J].J Transl Med,2021,19(1):96.
[26]	BAO C,LIU C,LIU Q,et al.Liproxstatin-1 alleviates LPS/IL-13-induced bronchial epithelial cell injury and neutrophilic asthma in mice by inhibiting ferroptosis[J].Int Immunopharmacol,2022,109:108770.
[27]	ZHENG H R,JI C N,ZOU X Q,et al.Molecular cloning and characterization of a novel human putative transmembrane protein homologous to mouse sideroflexin associated with sideroblastic anemia[J].DNA Seq,2003,14(5):369-373.
[28]	LI N,WANG W,ZHOU H,et al.Ferritinophagy-mediated ferroptosis is involved in sepsis-induced cardiac injury[J].Free Radical Biol Med,2020,160:303-318.
[29]	FANG J,KONG B,SHUAI W,et al.Ferroportin-mediated ferroptosis involved in new-onset atrial fibrillation with LPS-induced endotoxemia[J].Eur J Pharmacol,2021,913:174622.
[30]	KUANG F M,LIU J,TANG D L,et al.Oxidative damage and antioxidant defense in ferroptosis[J].Front Cell Dev Biol,2020,8:586578.
[31]	ZHU G Q,SUI S P,SHI F Y,et al.Inhibition of USP14 suppresses ferroptosis and inflammation in LPS-induced goat mammary epithelial cells through ubiquitylating the IL-6 protein[J].Hereditas,2022,159(1):21.
[32]	LUO H S,ZHANG R.Icariin enhances cell survival in lipopolysaccharide-induced synoviocytes by suppressing ferroptosis via the Xc-/GPX4 axis[J].Exp Ther Med,2020,21(1):72.
[33]	YANG M X,LU Z J,LI F L,et al.Escherichia coli induced ferroptosis in red blood cells of grass carp (Ctenopharyngodon idella)[J].Fish Shellfish Immun,2021,112:159-167.
[34]	PENARANDA C,CHUMBLER N M,HUNG D T.Dual transcriptional analysis reveals adaptation of host and pathogen to intracellular survival of Pseudomonas aeruginosa associated with urinary tract infection[J].PLoS Pathog,2021,17(4):e1009534.
[35]	ZHU H,SANTO A,JIA Z Q,et al.GPx4 in bacterial infection and polymicrobial sepsis:Involvement of ferroptosis and pyroptosis[J].React Oxyg Species (Apex),2019,7(21):154-160.
[36]	DAR H H,TYURINA Y Y,MIKULSKA-RUMINSKA K,et al.Pseudomonas aeruginosa utilizes host polyunsaturated phosphatidylethanolamines to trigger theft-ferroptosis in bronchial epithelium[J].J Clin Invest,2018,128(10):4639-4653.
[37]	OUSINGSAWAT J,SCHREIBER R,GULBINS E,et al.P. aeruginosa induced lipid peroxidation causes ferroptotic cell death in airways[J].Cell Physiol Biochem,2021,55(5):590-604.
[38]	AMARAL E P,COSTA D L,NAMASIVAYAM S,et al.A major role for ferroptosis in Mycobacterium tuberculosis-induced cell death and tissue necrosis[J].J Exp Med,2019,216(3):556-570.
[39]	SOE Y M,BEDOUI S,STINEAR T P,et al.Intracellular Staphylococcus aureus and host cell death pathways[J].Cell Microbiol,2021,23(5):e13317.
[40]	CHRISTMAS B A F,ROLFE M D,ROSE M,et al.Staphylococcus aureus adaptation to aerobic low-redox-potential environments:Implications for an intracellular lifestyle[J].Microbiology (Reading),2019,165(7):779-791.
[41]	TORRES V J,ATTIA A S,MASON W J,et al.Staphylococcus aureus fur regulates the expression of virulence factors that contribute to the pathogenesis of pneumonia[J].Infect Immun,2010,78(4):1618-1628.
[42]	WANG F,WANG R L,LIU H F.The acute pulmonary toxicity in mice induced by Staphylococcus aureus,particulate matter,and their combination[J].Exp Anim,2019,68(2):159-168.
[43]	BEAVERS W N,MONTEITH A J,AMARNATH V,et al.Arachidonic acid kills Staphylococcus aureus through a lipid peroxidation mechanism[J].mBio,2019,10(5):e01333-19.
[44]	LIU Q J,WU J,ZHANG X F,et al.Iron homeostasis and disorders revisited in the sepsis[J].Free Radical Biol Med,2021,165:1-13.
[45]	COLINS A,GERDTZEN Z P,NUÑEZ M T,et al.Mathematical modeling of intestinal iron absorption using genetic programming[J].PLoS One,2017,12(1):e0169601.
[46]	DONOVAN A,BROWNLIE A,ZHOU Y,et al.Positional cloning of zebrafish ferroportin1 identifies a conserved vertebrate iron exporter[J].Nature,2000,403(6771):776-781.
[47]	LUDWICZEK S,AIGNER E,THEURL I,et al.Cytokine-mediated regulation of iron transport in human monocytic cells[J].Blood,2003,101(10):4148-4154.
[48]	WEI S S,BI J B,YANG L F,et al.Serum irisin levels are decreased in patients with sepsis,and exogenous irisin suppresses ferroptosis in the liver of septic mice[J].Clin Transl Med,2020,10(5):e173.
[49]	WANG Q,DU F,QIAN Z M,et al.Lipopolysaccharide induces a significant increase in expression of iron regulatory hormone Hepcidin in the cortex and substantia Nigra in rat brain[J].Endocrinology,2008,149(8):3920-3925.
[50]	URRUTIA P,AGUIRRE P,ESPARZA A,et al.Inflammation alters the expression of DMT1,FPN1 and hepcidin,and it causes iron accumulation in central nervous system cells[J].J Neurochem,2013,126(4):541-549.
[51]	WESSELS I,COUSINS R J.Zinc dyshomeostasis during polymicrobial sepsis in mice involves zinc transporter Zip14 and can be overcome by zinc supplementation[J].Am J Physiol Gastrointest Liver Physiol,2015,309(9):G768-G778.
[52]	AYDEMIR T B,COUSINS R J.The multiple faces of the metal transporter ZIP14(SLC39A14)[J].J Nutr,2018,148(2):174-184.
[53]	SANCHEZ M,GALY B,SCHWANHAEUSSER B,et al.Iron regulatory protein-1 and -2:Transcriptome-wide definition of binding mRNAs and shaping of the cellular proteome by iron regulatory proteins[J].Blood,2011,118(22):E168-E179.
[54]	MANCIAS J D,VAITES L P,NISSIM S,et al.Ferritinophagy via NCOA4 is required for erythropoiesis and is regulated by iron dependent HERC2-mediated proteolysis[J].Elife,2015,4:e10308.
[55]	HUANG W X,ZHAN Y L,ZHENG Y F,et al.Up-regulated ferritin in periodontitis promotes inflammatory cytokine expression in human periodontal ligament cells through transferrin receptor via ERK/P38 MAPK pathways[J].Clin Sci (Lond),2019,133(1):135-148.
[56]	GUO W,ZHAO Y H,LI H X,et al.NCOA4-mediated ferritinophagy promoted inflammatory responses in periodontitis[J].J Periodontal Res,2021,56(3):523-534.
[57]	MAYR L,GRABHERR F,SCHWÄRZLER J,et al.Dietary lipids fuel GPX4-restricted enteritis resembling Crohn's disease[J].Nat Commun,2020,11(1):1775.
[58]	FUHRMANN D C,MONDORF A,BEIFUSS J,et al.Hypoxia inhibits ferritinophagy,increases mitochondrial ferritin,and protects from ferroptosis[J].Redox Biol,2020,36:101670.
[59]	GAO G F,LI J,ZHANG Y T,et al.Cellular iron metabolism and regulation[J].Adv Exp Med Biol,2019,1173:21-32.
[60]	GUIDA C,ALTAMURA S,KLEIN F A,et al.A novel inflammatory pathway mediating rapid hepcidin-independent hypoferremia[J].Blood,2015,125(14):2265-2275.
[61]	PALANISAMY G S,KIRK N M,ACKART D F,et al.Evidence for oxidative stress and defective antioxidant response in guinea pigs with tuberculosis[J].PLoS One,2011,6(10):e26254.
[62]	MARRO S,CHIABRANDO D,MESSANA E,et al. Heme controls ferroportin1(FPN1) transcription involving Bach1,Nrf2 and a MARE/ARE sequence motif at position-7007 of the FPN1 promoter[J].Haematologica,2010,95(8):1261-1268.
[63]	TANG D L,CHEN X,KANG R,et al.Ferroptosis:molecular mechanisms and health implications[J].Cell Res,2021,31(2):107-125.
[64]	HADIAN K,STOCKWELL B R.A roadmap to creating ferroptosis-based medicines[J].Nat Chem Biol,2021,17(11):1113-1116.
[65]	KACZOROWSKI Ł,POWIERSKA-CZARNY J,WOLKO Ł,et al.The influence of bacteria causing subclinical mastitis on the structure of the cow's milk microbiome[J].Molecules,2022,27(6):1829.
[66]	SIQUEIRA L C,FAVARETTO B,MORAES B T,et al.Bovine Endometritis and the inflammatory peripheral cholinergic system[J].Appl Biochem Biotechnol,2020,190(4):1242-1256.
[67]	MAVANGIRA V,KUHN M J,ABUELO A,et al.Activity of sEH and oxidant status during systemic bovine coliform mastitis[J].Antioxidants (Basel),2021,10(5):812.
[68]	LAUZON K,ZHAO X,LACASSE P.Deferoxamine reduces tissue damage during endotoxin-induced mastitis in dairy cows[J].J Dairy Sci,2006,89(10):3846-3857.
[69]	CHEN P,YANG J Y,WU N W,et al.Streptococcus lutetiensis induces autophagy via oxidative stress in bovine mammary epithelial cells[J].Oxid Med Cell Longev,2022,2022:2549772.
[70]	GUGLIANDOLO E,FUSCO R,LICATA P,et al.Protective effect of hydroxytyrosol on LPS-induced inflammation and oxidative stress in bovine endometrial epithelial cell line[J].Vet Sci,2020,7(4):161.

铁死亡在细菌性感染中的研究进展

Research Progress of Ferroptosis in Bacterial Infection

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