Background The aim of this study was to investigate the role of necroptosis in deoxynivalenol(DON)-induced liver injury and inflammation in weaned piglets.Methods In Exp.1,12 weaned piglets were divided into 2 groups ...Background The aim of this study was to investigate the role of necroptosis in deoxynivalenol(DON)-induced liver injury and inflammation in weaned piglets.Methods In Exp.1,12 weaned piglets were divided into 2 groups including pigs fed basal diet and pigs fed diet contaminated with 4 mg/kg DON for 21 d.In Exp.2,12 weaned piglets were divided into 2 groups including con-trol piglets and piglets given a gavage of 2 mg/kg body weight(BW)DON.In Exp.3,24 weaned piglets were used in a 2×2 factorial design and the main factors including necrostatin-1(Nec-1)(DMSO or 0.5 mg/kg BW Nec-1)and DON challenge(saline or 2 mg/kg BW DON gavage).On 21 d in Exp.1,or at 6 h post DON gavage in Exp.2 and 3,pigs were killed for blood samples and liver tissues.Liver histology,blood biochemical indicators,and liver inflamma-tion and necroptosis signals were tested.Results Dietary or oral gavage with DON caused liver morphological damage in piglets.Dietary DON led to hepato-cyte damage indicated by increased aspartate transaminase(AST)activity and AST/alanine aminotransferase(ALT)ratio,and DON gavage also caused hepatocyte damage and cholestasis indicated by increased AST and alkaline phosphatase(AKP)activities.Dietary DON caused liver necroptosis indicated by increased protein abundance of total receptor interacting protein kinase 3(t-RIP3)and total mixed lineage kinase domain-like protein(t-MLKL).Moreover,DON gavage increased mRNA expression of interleukin(IL)-6 and IL-1βin liver.DON gavage also induced liver necroptosis demonstrated by increased protein abundance of t-RIP3,phosphorylated-RIP3(p-RIP3),t-MLKL and p-MLKL.However,pretreatment with Nec-1,a specific inhibitor of necroptosis,inhibited liver necroptosis indi-cated by decreased protein expression of t-RIP3,p-RIP3,t-MLKL and p-MLKL.Nec-1 pretreatment reduced liver morphological damage after DON gavage.Pretreatment with Nec-1 also attenuated liver damage induced by DON indicated by decreased activities of AST and AKP.Furthermore,Nec-1 pretreatment inhibited liver mRNA expression of IL-6 and IL-1βafter DON challenge.Conclusions Our data demonstrate for the first time that necroptosis contributes to DON-induced liver injury and inflammation in piglets.展开更多
Background:Necroptosis and pyroptosis are newly identified forms of programmed cell death,which play a vital role in development of many gastrointestinal disorders.Although plant polyphenols have been reported to prot...Background:Necroptosis and pyroptosis are newly identified forms of programmed cell death,which play a vital role in development of many gastrointestinal disorders.Although plant polyphenols have been reported to protect intestinal health,it is still unclear whether there is a beneficial role of plant polyphenols in modulating necroptosis and pyroptosis in intestinal porcine epithelial cell line(IPEC-1)infected with enterotoxigenic Escherichia coli(ETEC)K88.This research was conducted to explore whether plant polyphenols including protocatechuic acid(PCA)and quercetin(Que),attenuated inflammation and injury of IPEC-1 caused by ETEC K88 through regulating necroptosis and pyroptosis signaling pathways.Methods:IPEC-1 cells were treated with PCA(40μmol/L)or Que(10μmol/L)in the presence or absence of ETEC K88.Results:PCA and Que decreased ETEC K88 adhesion and endotoxin level(P<0.05)in cell supernatant.PCA and Que increased cell number(P<0.001)and decreased lactate dehydrogenases(LDH)activity(P<0.05)in cell supernatant after ETEC infection.PCA and Que improved transepithelial electrical resistance(TEER)(P<0.001)and reduced fluorescein isothiocyanate-labeled dextran(FD4)flux(P<0.001),and enhanced membrane protein abundance of occludin,claudin-1 and ZO-1(P<0.05),and rescued distribution of these tight junction proteins(P<0.05)after ETEC infection.PCA and Que also declined cell necrosis ratio(P<0.05).PCA and Que reduced mRNA abundance and concentration of tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-8(P<0.001),and down-regulated gene expression of toll-like receptors 4(TLR4)and its downstream signals(P<0.001)after ETEC infection.PCA and Que down-regulated protein abundance of total receptor interacting protein kinase 1(t-RIP1),phosphorylated-RIP1(p-RIP1),p-RIP1/t-RIP1,t-RIP3,p-RIP3,mixed lineage kinase domain-like protein(MLKL),p-MLKL,dynamin-related protein 1(DRP1),phosphoglycerate mutase 5(PGAM5)and high mobility group box 1(HMGB1)(P<0.05)after ETEC infection.Moreover,PCA and Que reduced protein abundance of nod-like receptor protein 3(NLRP3),nod-like receptors family CARD domain-containing protein 4(NLRC4),apoptosis-associated speck-like protein containing a CARD(ASC),gasdermin D(GSDMD)and caspase-1(P<0.05)after ETEC infection.Conclusions:In general,our data suggest that PCA and Que are capable of attenuating ETEC-caused intestinal inflammation and damage via inhibiting necroptosis and pyroptosis signaling pathways.展开更多
基金Project of National Natural Science Foundation of China(U22A20517 and 32272906).
文摘Background The aim of this study was to investigate the role of necroptosis in deoxynivalenol(DON)-induced liver injury and inflammation in weaned piglets.Methods In Exp.1,12 weaned piglets were divided into 2 groups including pigs fed basal diet and pigs fed diet contaminated with 4 mg/kg DON for 21 d.In Exp.2,12 weaned piglets were divided into 2 groups including con-trol piglets and piglets given a gavage of 2 mg/kg body weight(BW)DON.In Exp.3,24 weaned piglets were used in a 2×2 factorial design and the main factors including necrostatin-1(Nec-1)(DMSO or 0.5 mg/kg BW Nec-1)and DON challenge(saline or 2 mg/kg BW DON gavage).On 21 d in Exp.1,or at 6 h post DON gavage in Exp.2 and 3,pigs were killed for blood samples and liver tissues.Liver histology,blood biochemical indicators,and liver inflamma-tion and necroptosis signals were tested.Results Dietary or oral gavage with DON caused liver morphological damage in piglets.Dietary DON led to hepato-cyte damage indicated by increased aspartate transaminase(AST)activity and AST/alanine aminotransferase(ALT)ratio,and DON gavage also caused hepatocyte damage and cholestasis indicated by increased AST and alkaline phosphatase(AKP)activities.Dietary DON caused liver necroptosis indicated by increased protein abundance of total receptor interacting protein kinase 3(t-RIP3)and total mixed lineage kinase domain-like protein(t-MLKL).Moreover,DON gavage increased mRNA expression of interleukin(IL)-6 and IL-1βin liver.DON gavage also induced liver necroptosis demonstrated by increased protein abundance of t-RIP3,phosphorylated-RIP3(p-RIP3),t-MLKL and p-MLKL.However,pretreatment with Nec-1,a specific inhibitor of necroptosis,inhibited liver necroptosis indi-cated by decreased protein expression of t-RIP3,p-RIP3,t-MLKL and p-MLKL.Nec-1 pretreatment reduced liver morphological damage after DON gavage.Pretreatment with Nec-1 also attenuated liver damage induced by DON indicated by decreased activities of AST and AKP.Furthermore,Nec-1 pretreatment inhibited liver mRNA expression of IL-6 and IL-1βafter DON challenge.Conclusions Our data demonstrate for the first time that necroptosis contributes to DON-induced liver injury and inflammation in piglets.
基金provided by National Key R&D Program of China(2022YFD1300403)National Natural Science Foundation of China(No.U22A20517,32272906,and 31802070)Wuhan Science and Technology Bureau(No.2022020801010391)。
文摘Background:Necroptosis and pyroptosis are newly identified forms of programmed cell death,which play a vital role in development of many gastrointestinal disorders.Although plant polyphenols have been reported to protect intestinal health,it is still unclear whether there is a beneficial role of plant polyphenols in modulating necroptosis and pyroptosis in intestinal porcine epithelial cell line(IPEC-1)infected with enterotoxigenic Escherichia coli(ETEC)K88.This research was conducted to explore whether plant polyphenols including protocatechuic acid(PCA)and quercetin(Que),attenuated inflammation and injury of IPEC-1 caused by ETEC K88 through regulating necroptosis and pyroptosis signaling pathways.Methods:IPEC-1 cells were treated with PCA(40μmol/L)or Que(10μmol/L)in the presence or absence of ETEC K88.Results:PCA and Que decreased ETEC K88 adhesion and endotoxin level(P<0.05)in cell supernatant.PCA and Que increased cell number(P<0.001)and decreased lactate dehydrogenases(LDH)activity(P<0.05)in cell supernatant after ETEC infection.PCA and Que improved transepithelial electrical resistance(TEER)(P<0.001)and reduced fluorescein isothiocyanate-labeled dextran(FD4)flux(P<0.001),and enhanced membrane protein abundance of occludin,claudin-1 and ZO-1(P<0.05),and rescued distribution of these tight junction proteins(P<0.05)after ETEC infection.PCA and Que also declined cell necrosis ratio(P<0.05).PCA and Que reduced mRNA abundance and concentration of tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-8(P<0.001),and down-regulated gene expression of toll-like receptors 4(TLR4)and its downstream signals(P<0.001)after ETEC infection.PCA and Que down-regulated protein abundance of total receptor interacting protein kinase 1(t-RIP1),phosphorylated-RIP1(p-RIP1),p-RIP1/t-RIP1,t-RIP3,p-RIP3,mixed lineage kinase domain-like protein(MLKL),p-MLKL,dynamin-related protein 1(DRP1),phosphoglycerate mutase 5(PGAM5)and high mobility group box 1(HMGB1)(P<0.05)after ETEC infection.Moreover,PCA and Que reduced protein abundance of nod-like receptor protein 3(NLRP3),nod-like receptors family CARD domain-containing protein 4(NLRC4),apoptosis-associated speck-like protein containing a CARD(ASC),gasdermin D(GSDMD)and caspase-1(P<0.05)after ETEC infection.Conclusions:In general,our data suggest that PCA and Que are capable of attenuating ETEC-caused intestinal inflammation and damage via inhibiting necroptosis and pyroptosis signaling pathways.
