该文主要探究汉黄芩素(WOG)调控STING/NF-κB通路对M5诱导的银屑病样HaCaT细胞模型炎症反应的影响。将HaCaT细胞随机分为NC组、五联因子(M5)组、M5+WOG组、M5+STING抑制剂(H-151)组、M5+WOG+STING激动剂(SR-717)组。采用CCK-8、EdU染色...该文主要探究汉黄芩素(WOG)调控STING/NF-κB通路对M5诱导的银屑病样HaCaT细胞模型炎症反应的影响。将HaCaT细胞随机分为NC组、五联因子(M5)组、M5+WOG组、M5+STING抑制剂(H-151)组、M5+WOG+STING激动剂(SR-717)组。采用CCK-8、EdU染色法和流式细胞仪分别检测细胞增殖和凋亡情况;免疫荧光法检测细胞屏障功能损伤相关指标;qPCR和ELISA分别检测炎症反应和银屑病特征性因子表达情况;Western blot检测STING/NF-κB信号通路相关蛋白表达情况。与NC组比较,M5组Ha Ca T细胞增殖活性,增殖率,IL-6、IL-8及TNF-αmRNA相对表达量,S100A7、S100A8及DEFB4含量,以及p-STING/STING、p-TBK1/TBK1、p-p65/p65值均上调,凋亡率以及ZO-1、Occludin及E-cadherin的阳性率均下调(P<0.05)。而WOG或STING抑制剂H-151的处理则有效逆转了上述M5诱导的效应(P<0.05)。值得注意的是,STING激动剂SR-717可部分抵消WOG的保护作用,表明WOG的疗效依赖于对STING/NF-κB通路的抑制(P<0.05)。WOG通过抑制STING/NF-κB信号通路有效抑制M5诱导的银屑病样Ha Ca T细胞增殖,促进其凋亡,修复屏障功能损伤,并减轻炎症反应。展开更多
目的:探讨干扰素基因刺激因子(stimulator of interferon genes,STING)介导的铁死亡在有氧运动改善高脂饮食小鼠心肌损伤中的作用及机制。方法:雄性C57BL/6J小鼠采用高脂饮食喂养,进行8周有氧跑台运动,通过腹腔注射diABZI药理学激活STIN...目的:探讨干扰素基因刺激因子(stimulator of interferon genes,STING)介导的铁死亡在有氧运动改善高脂饮食小鼠心肌损伤中的作用及机制。方法:雄性C57BL/6J小鼠采用高脂饮食喂养,进行8周有氧跑台运动,通过腹腔注射diABZI药理学激活STING、尾静脉注射腺相关病毒9型敲低心肌STING。体外实验中,采用棕榈酸(palmitic acid,PA)处理大鼠H9C2心肌细胞,模拟体外高脂诱导的心肌细胞损伤,通过小干扰RNA敲低STING,小分子抑制剂RSL3干预抑制谷胱甘肽过氧化物酶4(glutathione Peroxidase 4,GPX4)表达。小动物超声成像系统检测小鼠心功能;组织学染色分析心肌组织病理变化;透射电镜观察心肌线粒体形态结构;试剂盒检测小鼠心肌组织和H9C2细胞亚铁离子(Fe2+)、丙二醛(malondialdehyde,MDA)、总超氧化物歧化酶(total superoxide dismutase,T-SOD)、谷胱甘肽(glutathione,GSH)和氧化型谷胱甘肽(oxidized glutathione,GSSG)水平;二氢乙啶(dihydroethidium,DHE)染色评估活性氧(reative oxygen species,ROS)水平;JC-1染色检测线粒体膜电位变化;Western blot检测STING、GPX4、溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11)和酰基辅酶A合成酶长链家族成员4(acyl-CoA synthetase long-chain family member 4,ACSL4)蛋白表达。结果:(1)有氧运动显著抑制高脂饮食小鼠心肌STING表达。(2)STING激动剂di ABZI显著削弱有氧运动对高脂饮食小鼠心肌损伤的改善作用和对铁死亡的抑制作用。(3)敲低STING显著改善高脂饮食小鼠心脏功能和结构损伤,抑制心肌铁死亡。(4)敲低STING显著抑制PA诱导的H9C2细胞铁死亡,而GPX4抑制剂RSL3则削弱STING敲低对铁死亡的抑制作用。结论:STING-GPX4轴在高脂诱导的心肌细胞铁死亡中发挥着重要作用。有氧运动可通过下调STING表达,抑制铁死亡,改善高脂饮食小鼠心肌损伤。展开更多
The cyclic guanosine monophosphate-adenosine monophosphate synthase and the stimulator of interferon genes(cGAS-STING)has emerged as a promising target for cancer immunotherapy.However,the development of natural STING...The cyclic guanosine monophosphate-adenosine monophosphate synthase and the stimulator of interferon genes(cGAS-STING)has emerged as a promising target for cancer immunotherapy.However,the development of natural STING agonists is impeded by several challenges,including limited biostability,poor pharmacokinetics,and inefficient cytosolic delivery.Herein,we meticulously designed a doublelayer polyethylenimine(PEI)modified nanoscale covalent organic polymer(CPGP)for efficient delivery of 23cyclic guanosine monophosphate-adenosine monophosphate(cGAMP),a natural STING agonist.The double-layer PEI structured CPGP enhanced both the loading capacity and stability of cGAMP.Furthermore,CPGP improved the intracellular delivery efficiency and amplified the activation of STING pathway for the secretion of type-I interferon and pro-inflammatory cytokines.In contrast,single-layered nanoparticles failed to permit stable loading and intracellular delivery of cGAMP for immune response.The nano-STING agonist also mitigated the immunosuppressive tumor microenvironment(TME)by reducing regulatory T cells and polarizing M2 macrophages to the M1 phenotype,thereby creating an immune-supportive TME to enhance adaptive immune responses.The combination of CPGP and immune checkpoint blockers showed synergistic effect,further enhancing the inhibition effect on tumor growth.This double-layer PEI modified CPGP may offer a generalizable platform for other natural dinucleotide STING agonists to overcome the cascade delivery barriers,augmenting immune activation for tumor immunotherapy.展开更多
Background:The study of autoinflammatory diseases has uncovered mechanisms underlying cytokine dysregulation and inflammation.Methods:We analyzed the DNA of an index patient with early-onset systemic inflammation,cuta...Background:The study of autoinflammatory diseases has uncovered mechanisms underlying cytokine dysregulation and inflammation.Methods:We analyzed the DNA of an index patient with early-onset systemic inflammation,cutaneous vasculopathy,and pulmonary inflammation.We sequenced a candidate gene,TMEM173,encoding the stimulator of interferon genes(STING),in this patient and in five unrelated children with similar clinical phenotypes.Four children were evaluated clinically and immunologically.With the STING ligand cyclic guanosine monophosphate-adenosine monophosphate(cGAMP),we stimulated peripheral-blood mononuclear cells and fibroblasts from patients and controls,as well as commercially obtained endothelial cells,and then assayed transcription of IFNB1,the gene encoding interferon-β,in the stimulated cells.We analyzed IFNB1 reporter levels in HEK293T cells cotransfected with mutant or nonmutant STING constructs.Mutant STING leads to increased phosphorylation of signal transducer and activator of transcription 1(STAT1),so we tested the effect of Janus kinase(JAK)inhibitors on STAT1 phosphorylation in lymphocytes from the affected children and controls.展开更多
Background:The study of autoinflammatory diseases has uncovered mechanisms underlying cytokine dysregulation and inflammation.Methods:We analyzed the DNA of an index patient with early-onset systemic inflammation,cuta...Background:The study of autoinflammatory diseases has uncovered mechanisms underlying cytokine dysregulation and inflammation.Methods:We analyzed the DNA of an index patient with early-onset systemic inflammation,cutaneous vasculopathy,and pulmonary inflammation.We sequenced a candidate gene,TMEM173,encoding the stimulator of interferon genes(STING),in this patient and in five unrelated children with similar clinical phenotypes.展开更多
Objective:Clinical use of stimulator of interferon genes(STING)agonists has challenges due to poor responsiveness and variable efficacy.Therefore,identifying tumor types that are sensitive to these agents and clarifyi...Objective:Clinical use of stimulator of interferon genes(STING)agonists has challenges due to poor responsiveness and variable efficacy.Therefore,identifying tumor types that are sensitive to these agents and clarifying the underlying mechanisms are essential.Methods:In vitro screening was performed to identify tumor types that are sensitive to STING agonists.The non-nucleotide agonist,SR-717,and the macrocyclic agonist,E7766,were compared for efficacy.Complementary in vivo and in vitro studies,including gene-knockout models,HMGN2-knockout Neuro-2A and CT-2A cells apoptosis assays,and murine tumor models,were then performed.These experiments focused on the mechanism by which SR-717 mediates antitumor effects and emphasized the role of STING signaling-induced high-mobility group nucleosome-binding protein 2(HMGN2).In addition,the potential of HMGN2 as a prognostic biomarker was assessed.Results:Neuroblastomas and glioblastomas,two nervous system tumors,were shown to be sensitive to STING agonists.SR-717 exhibited greater antitumor efficacy compared to E7766.Mechanistic studies indicated that STING agonists promote apoptosis through activation of the intrinsic STING-signal transducer and activator of transcription 1(STAT1)-HMGN2 axis within tumor cells.Ectopic expression of HMGN2 in melanoma cells,which naturally lack HMGN2,led to significant apoptosis.Furthermore,analysis of The Cancer Genome Atlas and Gene Expression Omnibus databases revealed positive correlation between elevated HMGN2 expression and patient survival,supporting the utility of HMGN2 as a prognostic biomarker.Conclusions:This study clarified the mechanism underlying the potent antitumor activity of SR-717 in nervous system tumors through activation of the STING-STAT1-HMGN2 signaling pathway and demonstrated that SR-717 has superior efficacy compared to E7766.In addition,HMGN2 was shown to exhibit translational potential as a prognostic biomarker for patient survival.展开更多
This report presents a forensic evaluation of a case involving blindness(visual acuity grade 5)following a bee/wasp sting to the left eye.Through systematic analysis of the patient’s multiple hospital admissions,post...This report presents a forensic evaluation of a case involving blindness(visual acuity grade 5)following a bee/wasp sting to the left eye.Through systematic analysis of the patient’s multiple hospital admissions,postoperative follow-up data,and a review of the pathological mechanisms of ocular injury caused by bee venom,this study comprehensively assesses the injury characteristics,treatment course,and visual outcomes.Bee venom induces severe complications such as corneal damage,uveitis,cataract,and secondary glaucoma through multiple mechanisms including direct cytotoxicity,immune-inflammatory responses,and enzymatic hydrolysis.Despite interventions including anterior chamber irrigation,phacoemulsification with intraocular lens implantation,and antiglaucoma surgery,the affected eye ultimately lost light perception.Forensic examination confirmed the absence of light perception in the left eye and abnormal visual pathway function,consistent with clinical observations.According to the relevant Chinese disability assessment standard(JR/T 0083-2013,Article 4.2.2),the injury was classified as grade 7 disability.This study provides an in-depth discussion of the mechanisms and key forensic identification points in bee-sting-induced blindness,offering a scientific reference for similar forensic clinical cases.展开更多
文摘该文主要探究汉黄芩素(WOG)调控STING/NF-κB通路对M5诱导的银屑病样HaCaT细胞模型炎症反应的影响。将HaCaT细胞随机分为NC组、五联因子(M5)组、M5+WOG组、M5+STING抑制剂(H-151)组、M5+WOG+STING激动剂(SR-717)组。采用CCK-8、EdU染色法和流式细胞仪分别检测细胞增殖和凋亡情况;免疫荧光法检测细胞屏障功能损伤相关指标;qPCR和ELISA分别检测炎症反应和银屑病特征性因子表达情况;Western blot检测STING/NF-κB信号通路相关蛋白表达情况。与NC组比较,M5组Ha Ca T细胞增殖活性,增殖率,IL-6、IL-8及TNF-αmRNA相对表达量,S100A7、S100A8及DEFB4含量,以及p-STING/STING、p-TBK1/TBK1、p-p65/p65值均上调,凋亡率以及ZO-1、Occludin及E-cadherin的阳性率均下调(P<0.05)。而WOG或STING抑制剂H-151的处理则有效逆转了上述M5诱导的效应(P<0.05)。值得注意的是,STING激动剂SR-717可部分抵消WOG的保护作用,表明WOG的疗效依赖于对STING/NF-κB通路的抑制(P<0.05)。WOG通过抑制STING/NF-κB信号通路有效抑制M5诱导的银屑病样Ha Ca T细胞增殖,促进其凋亡,修复屏障功能损伤,并减轻炎症反应。
文摘目的:探讨干扰素基因刺激因子(stimulator of interferon genes,STING)介导的铁死亡在有氧运动改善高脂饮食小鼠心肌损伤中的作用及机制。方法:雄性C57BL/6J小鼠采用高脂饮食喂养,进行8周有氧跑台运动,通过腹腔注射diABZI药理学激活STING、尾静脉注射腺相关病毒9型敲低心肌STING。体外实验中,采用棕榈酸(palmitic acid,PA)处理大鼠H9C2心肌细胞,模拟体外高脂诱导的心肌细胞损伤,通过小干扰RNA敲低STING,小分子抑制剂RSL3干预抑制谷胱甘肽过氧化物酶4(glutathione Peroxidase 4,GPX4)表达。小动物超声成像系统检测小鼠心功能;组织学染色分析心肌组织病理变化;透射电镜观察心肌线粒体形态结构;试剂盒检测小鼠心肌组织和H9C2细胞亚铁离子(Fe2+)、丙二醛(malondialdehyde,MDA)、总超氧化物歧化酶(total superoxide dismutase,T-SOD)、谷胱甘肽(glutathione,GSH)和氧化型谷胱甘肽(oxidized glutathione,GSSG)水平;二氢乙啶(dihydroethidium,DHE)染色评估活性氧(reative oxygen species,ROS)水平;JC-1染色检测线粒体膜电位变化;Western blot检测STING、GPX4、溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11)和酰基辅酶A合成酶长链家族成员4(acyl-CoA synthetase long-chain family member 4,ACSL4)蛋白表达。结果:(1)有氧运动显著抑制高脂饮食小鼠心肌STING表达。(2)STING激动剂di ABZI显著削弱有氧运动对高脂饮食小鼠心肌损伤的改善作用和对铁死亡的抑制作用。(3)敲低STING显著改善高脂饮食小鼠心脏功能和结构损伤,抑制心肌铁死亡。(4)敲低STING显著抑制PA诱导的H9C2细胞铁死亡,而GPX4抑制剂RSL3则削弱STING敲低对铁死亡的抑制作用。结论:STING-GPX4轴在高脂诱导的心肌细胞铁死亡中发挥着重要作用。有氧运动可通过下调STING表达,抑制铁死亡,改善高脂饮食小鼠心肌损伤。
基金supported by the Beijing Natural Science Foundation(No.Z230021)the National Natural Science Foundation of China(No.52202356)+1 种基金the Non-profit Central Research Institute Fund of Chinese Academy of Medical Sciences(No.2021-RC350-001)the CAMS Innovation Fund for Medical Sciences(No.2022-I2M-1-013).
文摘The cyclic guanosine monophosphate-adenosine monophosphate synthase and the stimulator of interferon genes(cGAS-STING)has emerged as a promising target for cancer immunotherapy.However,the development of natural STING agonists is impeded by several challenges,including limited biostability,poor pharmacokinetics,and inefficient cytosolic delivery.Herein,we meticulously designed a doublelayer polyethylenimine(PEI)modified nanoscale covalent organic polymer(CPGP)for efficient delivery of 23cyclic guanosine monophosphate-adenosine monophosphate(cGAMP),a natural STING agonist.The double-layer PEI structured CPGP enhanced both the loading capacity and stability of cGAMP.Furthermore,CPGP improved the intracellular delivery efficiency and amplified the activation of STING pathway for the secretion of type-I interferon and pro-inflammatory cytokines.In contrast,single-layered nanoparticles failed to permit stable loading and intracellular delivery of cGAMP for immune response.The nano-STING agonist also mitigated the immunosuppressive tumor microenvironment(TME)by reducing regulatory T cells and polarizing M2 macrophages to the M1 phenotype,thereby creating an immune-supportive TME to enhance adaptive immune responses.The combination of CPGP and immune checkpoint blockers showed synergistic effect,further enhancing the inhibition effect on tumor growth.This double-layer PEI modified CPGP may offer a generalizable platform for other natural dinucleotide STING agonists to overcome the cascade delivery barriers,augmenting immune activation for tumor immunotherapy.
文摘Background:The study of autoinflammatory diseases has uncovered mechanisms underlying cytokine dysregulation and inflammation.Methods:We analyzed the DNA of an index patient with early-onset systemic inflammation,cutaneous vasculopathy,and pulmonary inflammation.We sequenced a candidate gene,TMEM173,encoding the stimulator of interferon genes(STING),in this patient and in five unrelated children with similar clinical phenotypes.Four children were evaluated clinically and immunologically.With the STING ligand cyclic guanosine monophosphate-adenosine monophosphate(cGAMP),we stimulated peripheral-blood mononuclear cells and fibroblasts from patients and controls,as well as commercially obtained endothelial cells,and then assayed transcription of IFNB1,the gene encoding interferon-β,in the stimulated cells.We analyzed IFNB1 reporter levels in HEK293T cells cotransfected with mutant or nonmutant STING constructs.Mutant STING leads to increased phosphorylation of signal transducer and activator of transcription 1(STAT1),so we tested the effect of Janus kinase(JAK)inhibitors on STAT1 phosphorylation in lymphocytes from the affected children and controls.
文摘Background:The study of autoinflammatory diseases has uncovered mechanisms underlying cytokine dysregulation and inflammation.Methods:We analyzed the DNA of an index patient with early-onset systemic inflammation,cutaneous vasculopathy,and pulmonary inflammation.We sequenced a candidate gene,TMEM173,encoding the stimulator of interferon genes(STING),in this patient and in five unrelated children with similar clinical phenotypes.
文摘Objective:Clinical use of stimulator of interferon genes(STING)agonists has challenges due to poor responsiveness and variable efficacy.Therefore,identifying tumor types that are sensitive to these agents and clarifying the underlying mechanisms are essential.Methods:In vitro screening was performed to identify tumor types that are sensitive to STING agonists.The non-nucleotide agonist,SR-717,and the macrocyclic agonist,E7766,were compared for efficacy.Complementary in vivo and in vitro studies,including gene-knockout models,HMGN2-knockout Neuro-2A and CT-2A cells apoptosis assays,and murine tumor models,were then performed.These experiments focused on the mechanism by which SR-717 mediates antitumor effects and emphasized the role of STING signaling-induced high-mobility group nucleosome-binding protein 2(HMGN2).In addition,the potential of HMGN2 as a prognostic biomarker was assessed.Results:Neuroblastomas and glioblastomas,two nervous system tumors,were shown to be sensitive to STING agonists.SR-717 exhibited greater antitumor efficacy compared to E7766.Mechanistic studies indicated that STING agonists promote apoptosis through activation of the intrinsic STING-signal transducer and activator of transcription 1(STAT1)-HMGN2 axis within tumor cells.Ectopic expression of HMGN2 in melanoma cells,which naturally lack HMGN2,led to significant apoptosis.Furthermore,analysis of The Cancer Genome Atlas and Gene Expression Omnibus databases revealed positive correlation between elevated HMGN2 expression and patient survival,supporting the utility of HMGN2 as a prognostic biomarker.Conclusions:This study clarified the mechanism underlying the potent antitumor activity of SR-717 in nervous system tumors through activation of the STING-STAT1-HMGN2 signaling pathway and demonstrated that SR-717 has superior efficacy compared to E7766.In addition,HMGN2 was shown to exhibit translational potential as a prognostic biomarker for patient survival.
文摘This report presents a forensic evaluation of a case involving blindness(visual acuity grade 5)following a bee/wasp sting to the left eye.Through systematic analysis of the patient’s multiple hospital admissions,postoperative follow-up data,and a review of the pathological mechanisms of ocular injury caused by bee venom,this study comprehensively assesses the injury characteristics,treatment course,and visual outcomes.Bee venom induces severe complications such as corneal damage,uveitis,cataract,and secondary glaucoma through multiple mechanisms including direct cytotoxicity,immune-inflammatory responses,and enzymatic hydrolysis.Despite interventions including anterior chamber irrigation,phacoemulsification with intraocular lens implantation,and antiglaucoma surgery,the affected eye ultimately lost light perception.Forensic examination confirmed the absence of light perception in the left eye and abnormal visual pathway function,consistent with clinical observations.According to the relevant Chinese disability assessment standard(JR/T 0083-2013,Article 4.2.2),the injury was classified as grade 7 disability.This study provides an in-depth discussion of the mechanisms and key forensic identification points in bee-sting-induced blindness,offering a scientific reference for similar forensic clinical cases.
