Macrophages are critical sentinel cells armed with multiple regulated necrosis pathways,including pyroptosis,apoptosis followed by secondary necrosis,and necroptosis,and are poised to undergo distinct form(s)of necrosis for tackling dangers of pathogenic infection or toxic exposure.The natural BH3-mimetic gossypol is a toxic phy-tochemical that can induce apoptosis and/or pyroptotic-like cell death,but what exact forms of regulated necrosis are induced remains largely unknown.Here we demonstrated that gossypol induces pyroptotic-like cell death in both unprimed and lipopolysaccharide-primed mouse bone marrow-derived macrophages(BMDMs),as evidenced by membrane swelling and ballooning accompanied by propidium iodide incorporation and lactic acid dehy-drogenase release.Notably,gossypol simultaneously induces the activation of both pyroptotic and apoptotic(followed by secondary necrosis)pathways but only weakly activates the necroptosis pathway.Unexpectedly,gossypol-induced necrosis is independent of nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3(NLRP3)inflammasome,as neither inhibitor for the NLRP3 pathway nor NLRP3 deficiency protects the macrophages from the necrosis.Furthermore,necrotic inhibitors or even pan-caspase inhibitor alone does not or only partly inhibit such necrosis.Instead,a combination of inhibitors composed of pan-caspase inhibitor IDN-6556,RIPK3 inhibitor GSK'872 and NADPH oxidase inhibitor GKT137831 not only markedly inhibits the ne-crosis,with all apoptotic and pyroptotic pathways being blocked,but also attenuates gossypol-induced peritonitis in mice.Lastly,the activation of the NLRP3 pathway and apoptotic caspase-3 appears to be independent of each other.Collectively,gossypol simultaneously induces the activation of multiple subroutines of regulated necrosis in macrophages depending on both apoptotic and inflammatory caspases.

Meiyan Zhong;Yuanting Huang;Bo Zeng;Lihui Xu;Chunsu Zhong;Jiahao Qiu;Xunjia Ye;Mingye Chen;Bo Hu;Dongyun Ouyang;Xianhui He

Department of Immunobiology,College of Life Science and Technology,Jinan University,Guangzhou 510632,China;Department of Cell Biology,College of Life Science and Technology,Jinan University,Guangzhou 510632,China;Department of Nephrology,the First Affiliated Hospital of Jinan University,Guangzhou 510632,China

生物化学与生物物理学报（英文版）

[1]Meiyan Zhong;Yuanting Huang;Bo Zeng;Lihui Xu;Chunsu Zhong;Jiahao Qiu;Xunjia Ye;Mingye Chen;Bo Hu;Dongyun Ouyang;Xianhui He-.Induction of multiple subroutines of regulated necrosis in murine macrophages by natural BH3-mimetic gossypol)[J].生物化学与生物物理学报（英文版）,2022(01):64-76

tochemical,pyroptotic,unprimed

Induction,multiple,subroutines,regulated,necrosis,murine,macrophages,by,natural,BH3,mimetic,gossypol,Macrophages,are,critical,sentinel,cells,armed,pathways,including,pyroptosis,apoptosis,followed,secondary,necroptosis,poised,undergo,distinct,tackling,dangers,pathogenic,infection,toxic,exposure,phy,that,can,like,death,but,what,exact,forms,induced,remains,largely,unknown,Here,demonstrated,induces,both,lipopolysaccharide,mouse,bone,marrow,derived,BMDMs,evidenced,membrane,swelling,ballooning,accompanied,propidium,iodide,incorporation,lactic,acid,dehy,drogenase,release,Notably,simultaneously,activation,apoptotic,only,weakly,activates,Unexpectedly,independent,nucleotide,binding,oligomerization,domain,receptor,family,pyrin,containing,NLRP3,inflammasome,neither,nor,deficiency,protects,from,Furthermore,necrotic,inhibitors,even,alone,does,not,partly,such,Instead,combination,composed,IDN,RIPK3,GSK,NADPH,oxidase,GKT137831,markedly,inhibits,being,blocked,also,attenuates,peritonitis,mice,Lastly,appears,each,other,Collectively,depending,inflammatory,caspases

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