Clinical studies have shown that Aggregatibacter actinomycetemcomitans(A.actinomycetemcomitans)is associated with aggressive periodontitis and can potentially trigger or exacerbate rheumatoid arthritis(RA).However,the...Clinical studies have shown that Aggregatibacter actinomycetemcomitans(A.actinomycetemcomitans)is associated with aggressive periodontitis and can potentially trigger or exacerbate rheumatoid arthritis(RA).However,the mechanism is poorly understood.Here,we show that systemic infection with A.actinomycetemcomitans triggers the progression of arthritis in mice anti-collagen antibody-induced arthritis(CAIA)model following IL-1βsecretion and cell infiltration in paws in a manner that is dependent on caspase-11-mediated inflammasome activation in macrophages.The administration of polymyxin B(PMB),chloroquine,and anti-CD11b antibody suppressed inflammasome activation in macrophages and arthritis in mice,suggesting that the recognition of lipopolysaccharide(LPS)in the cytosol after bacterial degradation by lysosomes and invasion via CD11b are needed to trigger arthritis following inflammasome activation in macrophages.These data reveal that the inhibition of caspase-11-mediated inflammasome activation potentiates aggravation of RA induced by infection with A.actinomycetemcomitans.This work highlights how RA can be progressed by inflammasome activation as a result of periodontitis-associated bacterial infection and discusses the mechanism of inflammasome activation in response to infection with A.actinomycetemcomitans.展开更多
目的对近年来脂多糖介导的炎症疾病含半胱氨酸的天冬氨酸蛋白水解酶11(cysteinyl aspartate specific proteinase-11,caspase-11)靶点药物进行综述。方法利用PubMed、中国知网等检索平台,检索脂多糖介导的不同炎症疾病中caspase-11靶向...目的对近年来脂多糖介导的炎症疾病含半胱氨酸的天冬氨酸蛋白水解酶11(cysteinyl aspartate specific proteinase-11,caspase-11)靶点药物进行综述。方法利用PubMed、中国知网等检索平台,检索脂多糖介导的不同炎症疾病中caspase-11靶向药物并进行了归纳汇总。结果根据文献查阅发现,若caspase-11激活,可诱发白介素1β、白介素18相关的炎症反应及Gasdermin D介导的细胞焦亡,从而导致大量炎症因子释放到胞外,引发强烈的炎症反应。caspase-11介导的炎症反应在帕金森病、阿尔兹海默症、肺纤维化、内毒素血症、脓毒症等疾病的发生发展中发挥了重要作用。结论对caspase-11靶点药物的探索,可能是未来脂多糖介导的相关炎症疾病药物研究的新方向。展开更多
Objective:To explore the protective effect and the underlying mechanism of silibinin(SIB),one of the active compounds from Silybum marianum(L.)Gaertn in endotoxemia.Methods:Mouse peritoneal macrophage were isolated vi...Objective:To explore the protective effect and the underlying mechanism of silibinin(SIB),one of the active compounds from Silybum marianum(L.)Gaertn in endotoxemia.Methods:Mouse peritoneal macrophage were isolated via intraperitoneally injection of BALB/c mice with thioglycolate medium.Cell viability was assessed using the cell counting kit-8,while cytotoxicity was determined through lactate dehydrogenase cytotoxicity assay.The protein expressions of interleukin(IL)-1α,IL-1β,and IL-18 were determined by enzyme-linked immunosorbent assay.Intracellular lipopolysaccharide(LPS)levels were measured by employing both the limulus amoebocyte lysate assay and flow cytometry.Additionally,proximity ligation assay was employed for the LPS and caspase-11 interaction.Mice were divided into 4 groups:the control,LPS,high-dose-SIB(100 mg/kg),and low-dose-SIB(100 mg/kg)groups(n=8).Zebrafish were divided into 4 groups:the control,LPS,high-dose-SIB(200μmol/L),and low-dose-SIB(100μmol/L)groups(n=30 for survival experiment and n=10 for gene expression analysis).The expression of caspase-11,gasdermin D(GSDMD),and N-GSDMD was determined by Western blot and the expressions of caspy2,gsdmeb,and IL-1βwere detected using quantitative real-time PCR.Histopathological observation was performed through hematoxylineosin staining,and protein levels in bronchoalveolar lavage fluid were quantified using the bicinchoninicacid protein assay.Results:SIB noticeably decreased caspase-11 and GSDMD-mediated pyroptosis and suppressed the secretion of IL-1α,IL-1β,and IL-18 induced by LPS(P<0.05).Moreover,SIB inhibited the translocation of LPS into the cytoplasm and the binding of caspase-11 and intracellular LPS(P<0.05).SIB also attenuated the expression of caspase-11 and N-terminal fragments of GSDMD,inhibited the relative cytokines,prolonged the survival time,and up-regulated the survival rate in the endotoxemia models(P<0.05).Conclusions:SIB can inhibit pyroptosis in the LPS-mediated endotoxemia model,at least in part,by inhibiting the caspase-11-mediated cleavage of GSDMD.Additionally,SIB inhibits the interaction of LPS and caspase-11 and inhibits the LPS-mediated up-regulation of caspase-11 expression,which relieves caspase-11-dependent cell pyroptosis and consequently attenuates LPS-mediated lethality.展开更多
Inflammatory caspase-11 senses and is activated by intracellular lipopolysaccharide(LPS)leading to pyroptosis that has critical role in defensing against bacterial infection,whereas its excess activation under pathoge...Inflammatory caspase-11 senses and is activated by intracellular lipopolysaccharide(LPS)leading to pyroptosis that has critical role in defensing against bacterial infection,whereas its excess activation under pathogenic circumstances may cause various inflammatory diseases.However,there are few known drugs that can control caspase-11 activation.We report here that scutellarin,a flavonoid from Erigeron breviscapus,acted as an inhibitor for caspase-11 activation in macrophages.Scutellarin dosedependently inhibited intracellular LPS-induced release of caspase-11 p26(indicative of caspase-11 activation)and generation of N-terminal fragment of gasdermin D(GSDMD-NT),leading to reduced pyroptosis.It also suppressed the activation of non-canonical nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3(NLRP3)inflammasome as evidenced by reduced apoptosisassociated speck-like protein containing a CARD(ASC)speck formation and decreased interleukin-1 beta(IL-1 b)and caspase-1 p10 secretion,whereas the NLRP3-specific inhibitor MCC950 only inhibited IL-1 b and caspase-1 p10 release and ASC speck formation but not pyroptosis.Scutellarin also suppressed LPS-induced caspase-11 activation and pyroptosis in RAW 264.7 cells lacking ASC expression.Moreover,scutellarin treatment increased Ser/Thr phosphorylation of caspase-11 at protein kinase A(PKA)-specific sites,and its inhibitory action on caspase-11 activation was largely abrogated by PKAinhibitor H89 or by adenylyl cyclase inhibitor MDL12330 A.Collectively,our data indicate that scutellarin inhibited caspase-11 activation and pyroptosis in macrophages at least partly via regulating the PKA signaling pathway.展开更多
目的 ：研究慢病毒载体GV115介导Caspase-3 si RNA转染人椎间盘髓核细胞的生物学效应。方法 ：采集12例外伤致脊柱爆裂性骨折患者（22~36岁）术中切除的椎间盘髓核组织,采用组织块法分离培养髓核细胞并传代。取第2代髓核细胞,分为GV115-...目的 ：研究慢病毒载体GV115介导Caspase-3 si RNA转染人椎间盘髓核细胞的生物学效应。方法 ：采集12例外伤致脊柱爆裂性骨折患者（22~36岁）术中切除的椎间盘髓核组织,采用组织块法分离培养髓核细胞并传代。取第2代髓核细胞,分为GV115-Caspase3 si RNA组、GV115组和对照组,各组12个细胞培养孔。在荧光显微镜下观察计数阳性髓核细胞,计算GV115-Caspase3 siRNA对人椎间盘髓核细胞的转染效率;采用免疫荧光法检测三组髓核细胞中Caspase-3表达;采用MTT法检测三组髓核细胞活性;采用Western-Blot法和Antonopulos法分别检测三组髓核细胞的Ⅱ型胶原和蛋白多糖含量。结果 ：椎间盘髓核细胞被成功分离培养,培养1周后细胞达到80%融合,进行传代培养。转染后1周（85.6±1.3）%的髓核细胞可被GV115-Caspase3 si RNA转染而表达绿色荧光素。GV115-Caspase3 siRNA组Caspase-3免疫荧光阳性细胞率[（19.4±3.2）%]较GV115组[（84.3±9.2）%]和对照组[（83.9±8.7）%]明显减少（P〈0.05）,OD值（1.56±0.21）较GV115组（0.91±0.15）和对照组（0.92±0.17）高（P〈0.05）,Ⅱ型胶原免疫印迹染色强度（1.32±0.09）较GV115组（0.81±0.05）和对照组（0.79±0.04）高（P〈0.05）,蛋白多糖含量（0.56±0.09）较GV115组（0.35±0.06）和对照组（0.34±0.05）高（P〈0.05）。结论：GV115-Caspase3 siRNA可高效转染人椎间盘髓核细胞,增强髓核细胞的生物活性,并促进细胞外基质的合成。展开更多
基金supported by the Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science(15H04730,16H05186,16K08772,17K12004,19H03467,21J00572)a contract research fund from the Japan Program for Infectious Diseases Research and Infrastructure for research on emerging and re-emerging infectious diseases provided by the Japan Agency for Medical Research and Development(AMED)the Project for Promoting Leading-edge Research in Oral Science at Tokyo Medical and Dental University。
文摘Clinical studies have shown that Aggregatibacter actinomycetemcomitans(A.actinomycetemcomitans)is associated with aggressive periodontitis and can potentially trigger or exacerbate rheumatoid arthritis(RA).However,the mechanism is poorly understood.Here,we show that systemic infection with A.actinomycetemcomitans triggers the progression of arthritis in mice anti-collagen antibody-induced arthritis(CAIA)model following IL-1βsecretion and cell infiltration in paws in a manner that is dependent on caspase-11-mediated inflammasome activation in macrophages.The administration of polymyxin B(PMB),chloroquine,and anti-CD11b antibody suppressed inflammasome activation in macrophages and arthritis in mice,suggesting that the recognition of lipopolysaccharide(LPS)in the cytosol after bacterial degradation by lysosomes and invasion via CD11b are needed to trigger arthritis following inflammasome activation in macrophages.These data reveal that the inhibition of caspase-11-mediated inflammasome activation potentiates aggravation of RA induced by infection with A.actinomycetemcomitans.This work highlights how RA can be progressed by inflammasome activation as a result of periodontitis-associated bacterial infection and discusses the mechanism of inflammasome activation in response to infection with A.actinomycetemcomitans.
文摘目的对近年来脂多糖介导的炎症疾病含半胱氨酸的天冬氨酸蛋白水解酶11(cysteinyl aspartate specific proteinase-11,caspase-11)靶点药物进行综述。方法利用PubMed、中国知网等检索平台,检索脂多糖介导的不同炎症疾病中caspase-11靶向药物并进行了归纳汇总。结果根据文献查阅发现,若caspase-11激活,可诱发白介素1β、白介素18相关的炎症反应及Gasdermin D介导的细胞焦亡,从而导致大量炎症因子释放到胞外,引发强烈的炎症反应。caspase-11介导的炎症反应在帕金森病、阿尔兹海默症、肺纤维化、内毒素血症、脓毒症等疾病的发生发展中发挥了重要作用。结论对caspase-11靶点药物的探索,可能是未来脂多糖介导的相关炎症疾病药物研究的新方向。
基金Supported by the Guangdong Basic and Applied Basic Research Foundation(No.2023A1515011106)Science and Technology Program of Guangzhou(No.2023A04J1826)。
文摘Objective:To explore the protective effect and the underlying mechanism of silibinin(SIB),one of the active compounds from Silybum marianum(L.)Gaertn in endotoxemia.Methods:Mouse peritoneal macrophage were isolated via intraperitoneally injection of BALB/c mice with thioglycolate medium.Cell viability was assessed using the cell counting kit-8,while cytotoxicity was determined through lactate dehydrogenase cytotoxicity assay.The protein expressions of interleukin(IL)-1α,IL-1β,and IL-18 were determined by enzyme-linked immunosorbent assay.Intracellular lipopolysaccharide(LPS)levels were measured by employing both the limulus amoebocyte lysate assay and flow cytometry.Additionally,proximity ligation assay was employed for the LPS and caspase-11 interaction.Mice were divided into 4 groups:the control,LPS,high-dose-SIB(100 mg/kg),and low-dose-SIB(100 mg/kg)groups(n=8).Zebrafish were divided into 4 groups:the control,LPS,high-dose-SIB(200μmol/L),and low-dose-SIB(100μmol/L)groups(n=30 for survival experiment and n=10 for gene expression analysis).The expression of caspase-11,gasdermin D(GSDMD),and N-GSDMD was determined by Western blot and the expressions of caspy2,gsdmeb,and IL-1βwere detected using quantitative real-time PCR.Histopathological observation was performed through hematoxylineosin staining,and protein levels in bronchoalveolar lavage fluid were quantified using the bicinchoninicacid protein assay.Results:SIB noticeably decreased caspase-11 and GSDMD-mediated pyroptosis and suppressed the secretion of IL-1α,IL-1β,and IL-18 induced by LPS(P<0.05).Moreover,SIB inhibited the translocation of LPS into the cytoplasm and the binding of caspase-11 and intracellular LPS(P<0.05).SIB also attenuated the expression of caspase-11 and N-terminal fragments of GSDMD,inhibited the relative cytokines,prolonged the survival time,and up-regulated the survival rate in the endotoxemia models(P<0.05).Conclusions:SIB can inhibit pyroptosis in the LPS-mediated endotoxemia model,at least in part,by inhibiting the caspase-11-mediated cleavage of GSDMD.Additionally,SIB inhibits the interaction of LPS and caspase-11 and inhibits the LPS-mediated up-regulation of caspase-11 expression,which relieves caspase-11-dependent cell pyroptosis and consequently attenuates LPS-mediated lethality.
基金supported by the National Natural Science Foundation of China(Nos.81773965,81873064,and 81673664)
文摘Inflammatory caspase-11 senses and is activated by intracellular lipopolysaccharide(LPS)leading to pyroptosis that has critical role in defensing against bacterial infection,whereas its excess activation under pathogenic circumstances may cause various inflammatory diseases.However,there are few known drugs that can control caspase-11 activation.We report here that scutellarin,a flavonoid from Erigeron breviscapus,acted as an inhibitor for caspase-11 activation in macrophages.Scutellarin dosedependently inhibited intracellular LPS-induced release of caspase-11 p26(indicative of caspase-11 activation)and generation of N-terminal fragment of gasdermin D(GSDMD-NT),leading to reduced pyroptosis.It also suppressed the activation of non-canonical nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3(NLRP3)inflammasome as evidenced by reduced apoptosisassociated speck-like protein containing a CARD(ASC)speck formation and decreased interleukin-1 beta(IL-1 b)and caspase-1 p10 secretion,whereas the NLRP3-specific inhibitor MCC950 only inhibited IL-1 b and caspase-1 p10 release and ASC speck formation but not pyroptosis.Scutellarin also suppressed LPS-induced caspase-11 activation and pyroptosis in RAW 264.7 cells lacking ASC expression.Moreover,scutellarin treatment increased Ser/Thr phosphorylation of caspase-11 at protein kinase A(PKA)-specific sites,and its inhibitory action on caspase-11 activation was largely abrogated by PKAinhibitor H89 or by adenylyl cyclase inhibitor MDL12330 A.Collectively,our data indicate that scutellarin inhibited caspase-11 activation and pyroptosis in macrophages at least partly via regulating the PKA signaling pathway.
文摘目的 ：研究慢病毒载体GV115介导Caspase-3 si RNA转染人椎间盘髓核细胞的生物学效应。方法 ：采集12例外伤致脊柱爆裂性骨折患者（22~36岁）术中切除的椎间盘髓核组织,采用组织块法分离培养髓核细胞并传代。取第2代髓核细胞,分为GV115-Caspase3 si RNA组、GV115组和对照组,各组12个细胞培养孔。在荧光显微镜下观察计数阳性髓核细胞,计算GV115-Caspase3 siRNA对人椎间盘髓核细胞的转染效率;采用免疫荧光法检测三组髓核细胞中Caspase-3表达;采用MTT法检测三组髓核细胞活性;采用Western-Blot法和Antonopulos法分别检测三组髓核细胞的Ⅱ型胶原和蛋白多糖含量。结果 ：椎间盘髓核细胞被成功分离培养,培养1周后细胞达到80%融合,进行传代培养。转染后1周（85.6±1.3）%的髓核细胞可被GV115-Caspase3 si RNA转染而表达绿色荧光素。GV115-Caspase3 siRNA组Caspase-3免疫荧光阳性细胞率[（19.4±3.2）%]较GV115组[（84.3±9.2）%]和对照组[（83.9±8.7）%]明显减少（P〈0.05）,OD值（1.56±0.21）较GV115组（0.91±0.15）和对照组（0.92±0.17）高（P〈0.05）,Ⅱ型胶原免疫印迹染色强度（1.32±0.09）较GV115组（0.81±0.05）和对照组（0.79±0.04）高（P〈0.05）,蛋白多糖含量（0.56±0.09）较GV115组（0.35±0.06）和对照组（0.34±0.05）高（P〈0.05）。结论：GV115-Caspase3 siRNA可高效转染人椎间盘髓核细胞,增强髓核细胞的生物活性,并促进细胞外基质的合成。
