目的检测分析异染色质蛋白1结合蛋白3(heterochromatin protein 1 binding protein 3,HP1BP3)在肝细胞癌中的表达及其对细胞恶性生物学行为的影响。方法应用TCGA数据库分析肝细胞癌与癌旁组织中HP1BP3基因的表达差异。生存检验分析HP1BP...目的检测分析异染色质蛋白1结合蛋白3(heterochromatin protein 1 binding protein 3,HP1BP3)在肝细胞癌中的表达及其对细胞恶性生物学行为的影响。方法应用TCGA数据库分析肝细胞癌与癌旁组织中HP1BP3基因的表达差异。生存检验分析HP1BP3表达与肝细胞癌患者总生存期的关系,单因素及多因素分析相关危险因素。应用Western blot和免疫组化法检测HP1BP3蛋白表达水平。用小干扰RNA(siRNA)分别转染Huh7、HepG2细胞,功能实验检测细胞增殖、迁移和侵袭能力。通过GSEA分析对其作用机制进行探究。结果TCGA数据库分析结果显示,HP1BP3基因在肝细胞癌组织中的表达明显高于非肿瘤组织(P<0.01),高表达组患者的总生存期较低表达组短(P=0.029)。HP1BP3基因是肝细胞癌的独立预测因子(P=0.024)。体外实验结果显示,HP1BP3基因在肝癌细胞株及肝细胞癌组织中表达上调。沉默HP1BP3基因可抑制肝癌细胞的增殖、侵袭和迁移。HP1BP3高表达与泛素化(P<0.001)、核酸切除修复(P<0.001)、RNA降解(P<0.001)、ERBB信号通路(P<0.001)、细胞周期(P<0.001)、细胞凋亡(P<0.001)通路有关。结论HP1BP3基因在肝细胞癌组织中表达上调,是肝细胞癌独立的预后危险因素,可促进肝细胞癌的发生、发展。展开更多
Autophagy plays a pivotal role in diverse biological processes,including the maintenance and differentiation of neural stem cells(NSCs).Interestingly,while complete deletion of Fip200 severely impairs NSC maintenance ...Autophagy plays a pivotal role in diverse biological processes,including the maintenance and differentiation of neural stem cells(NSCs).Interestingly,while complete deletion of Fip200 severely impairs NSC maintenance and differentiation,inhibiting canonical autophagy via deletion of core genes,such as Atg5,Atg16l1,and Atg7,or blockade of canonical interactions between FIP200 and ATG13(designated as FIP200-4A mutant or FIP200 KI)does not produce comparable detrimental effects.This highlights the likely critical involvement of the non-canonical functions of FIP200,the mechanisms of which have remained elusive.Here,utilizing genetic mouse models,we demonstrated that FIP200 mediates non-canonical autophagic degradation of p62/sequestome1,primarily via TAX1BP1 in NSCs.Conditional deletion of Tax1bp1 in fip200hGFAP conditional knock-in(cKI)mice led to NSC deficiency,resembling the fip200hGFAP conditional knockout(cKO)mouse phenotype.Notably,reintroducing wild-type TAX1BP1 not only restored the maintenance of NSCs derived from tax1bp1-knockout fip200hGFAP cKI mice but also led to a marked reduction in p62 aggregate accumulation.Conversely,a TAX1BP1 mutant incapable of binding to FIP200 or NBR1/p62 failed to achieve this restoration.Furthermore,conditional deletion of Tax1bp1 in fip200hGFAP cKO mice exacerbated NSC deficiency and p62 aggregate accumulation compared to fip200hGFAP cKO mice.Collectively,these findings illustrate the essential role of the FIP200-TAX1BP1 axis in mediating the non-canonical autophagic degradation of p62 aggregates towards NSC maintenance and function,presenting novel therapeutic targets for neurodegenerative diseases.展开更多
Acute myocardial infarction(MI),one of the most common cardiovascular emergencies,is a leading cause of morbidity and mortality.Ample evidence has revealed an essential role for inflammasome activation and autophagy i...Acute myocardial infarction(MI),one of the most common cardiovascular emergencies,is a leading cause of morbidity and mortality.Ample evidence has revealed an essential role for inflammasome activation and autophagy in the pathogenesis of acute MI.Tax1-binding protein 1(TAX1BP1),an adaptor molecule involved in termination of proinflammatory signaling,serves as an important selective autophagy adaptor,but its role in cardiac ischemia remains elusive.This study examined the role of TAX1BP1 in myocardial ischemic stress and the underlying mechanisms involved.Levels of TAX1BP1 were significantly downregulated in heart tissues of patients with ischemic heart disease and in a left anterior descending(LAD)ligation-induced model of acute MI.Adenovirus carrying TAX1BP1 was delivered into the myocardium.The acute MI induced procedure elicited an infarct and cardiac dysfunction,the effect of which was mitigated by TAX1BP1 overexpression with little effect from viral vector alone.TAX1BP1 nullified acute MI-induced activation of the NLRP3 inflammasome and associated mitochondrial dysfunction.TAX1BP1 overexpression suppressed NLRP3 mitochondrial localization by inhibiting the interaction of NLRP3 with mitochondrial antiviral signaling protein(MAVS).Further investigation revealed that ring finger protein 34(RNF34)was recruited to interact with TAX1BP1 thereby facilitating autophagic degradation of MAVS through K27-linked polyubiquitination of MAVS.Knockdown of RNF34 using siRNA nullified TAX1BP1 yielded protection against hypoxia-induced MAVS mitochondrial accumulation,NLRP3 inflammasome activation and associated loss of mitochondrial membrane potential.Taken together,our results favor a cardioprotective role for TAX1BP1 in acute MI through repression of inflammasome activation in a RNF34/MAVS-dependent manner.展开更多
Objective:The objective of this study was to determine the level of methotrexate(MTX)toxicity in the intestines of mice and to evaluate the protective effect of probiotics composed of Streptococcus,Bifidobacterium,and...Objective:The objective of this study was to determine the level of methotrexate(MTX)toxicity in the intestines of mice and to evaluate the protective effect of probiotics composed of Streptococcus,Bifidobacterium,and Lactobacillus species on intestinal cells during MTX treatment.Methods:Mice were divided into three groups:control,MTX group(received MTX injections),and MTX+probiotics group(received MTX injections along with a diet containing probiotics).Morphological and histological changes,the level of mitochondrial DNA(mtDNA)damage,the level of lipid peroxidation products,and gene expression in the mice’s small intestine were assessed.Results:We demonstrated that intraperitoneal MTX injections significantly increased mtDNA damage in the liver(p<0.001),small intestine(p<0.001),and blood of mice(p<0.01).MTX elevated the quantity of lipid peroxidation products in the liver and small intestine,indicating its strong prooxidative properties.MTX induced structural changes in the mice’s intestines,characterized by leukocytic infiltration of tissues.Probiotic therapy in mice partially mitigated the morphological and histological changes in the small intestine induced by MTX,reduced oxidative stress,and promoted increased expression of quinone oxidoreductase 1(Nqo1),which participates in both cell protection against oxidative stress and drug/xenobiotic detoxification.Probiotics prevented the upregulation of the proinflammatory cytokine IL-1b in the small intestine and induced increased expression of genes associated with the Nuclear factor erythroid 2-related factor 2/Antioxidant response element(Nrf2/ARE)pathway,an important mechanism of cell protection.Conclusions:Probiotics can be considered an effective approach to reducing the toxicity of MTX during psoriasis or cancer treatment.展开更多
文摘目的·扩增干扰素调节因子3(interferon regulator factor 3,IRF3)短发夹RNA(short hairpin RNA,sh RNA)腺病毒,并研究该病毒对脂多糖(lipopolysaccharide,LPS)刺激诱导Raw264.7细胞核内白介素受体相关激酶1结合蛋白1(interleukin-1 receptor associated kinase 1 binding protein 1,Irak1bp1)表达的影响。方法·IRF3 sh RNA腺病毒的扩增在人胚肾293 T(HEK293T)细胞中进行,并采用TCID 50法测定病毒滴度。Raw 264.7细胞随机分为4组,1组为腺病毒(-)LPS(-),2组为腺病毒(-)LPS(+),3组为腺病毒(+)LPS(-),4组为腺病毒(+)LPS(+)。细胞IRF3基因表达采用real-time PCR方法检测;核内IRF3及Irak1bp1的表达采用Western blotting方法检测。结果·经计算扩增腺病毒滴度为2.2×10^(11) PFU/m L,最佳MOI为300。LPS刺激后Raw 264.7细胞内IRF3 m RNA较对照组明显增加,核内IRF3蛋白及Irak1bp1表达也明显增加;IRF3 sh RNA腺病毒应用后,细胞对IRF3 m RNA的组成性表达及LPS刺激诱导的IRF3 m RNA和核内蛋白质表达均明显受抑,但未刺激状态下IRF3蛋白核内组成性表达无明显影响;IRF3 sh RNA腺病毒应用对细胞静息及LPS刺激诱导的核内Irak1bp1表达并无影响。结论·IRF3 sh RNA腺病毒能够有效抑制LPS刺激诱导的核内IRF3的表达,但并不影响核内Irak1bp1的表达。
文摘目的检测分析异染色质蛋白1结合蛋白3(heterochromatin protein 1 binding protein 3,HP1BP3)在肝细胞癌中的表达及其对细胞恶性生物学行为的影响。方法应用TCGA数据库分析肝细胞癌与癌旁组织中HP1BP3基因的表达差异。生存检验分析HP1BP3表达与肝细胞癌患者总生存期的关系,单因素及多因素分析相关危险因素。应用Western blot和免疫组化法检测HP1BP3蛋白表达水平。用小干扰RNA(siRNA)分别转染Huh7、HepG2细胞,功能实验检测细胞增殖、迁移和侵袭能力。通过GSEA分析对其作用机制进行探究。结果TCGA数据库分析结果显示,HP1BP3基因在肝细胞癌组织中的表达明显高于非肿瘤组织(P<0.01),高表达组患者的总生存期较低表达组短(P=0.029)。HP1BP3基因是肝细胞癌的独立预测因子(P=0.024)。体外实验结果显示,HP1BP3基因在肝癌细胞株及肝细胞癌组织中表达上调。沉默HP1BP3基因可抑制肝癌细胞的增殖、侵袭和迁移。HP1BP3高表达与泛素化(P<0.001)、核酸切除修复(P<0.001)、RNA降解(P<0.001)、ERBB信号通路(P<0.001)、细胞周期(P<0.001)、细胞凋亡(P<0.001)通路有关。结论HP1BP3基因在肝细胞癌组织中表达上调,是肝细胞癌独立的预后危险因素,可促进肝细胞癌的发生、发展。
基金National Natural Science Foundation of China(U2004138,81773132,81820108021)University Excellent Teaching Team of“Qinglan Project”in Jiangsu Province(2022-25)+1 种基金Henan Province Key Research and Development Project(232102521028)Excellent Youth Foundation of Henan Scientific Committee(21230040016)。
文摘Autophagy plays a pivotal role in diverse biological processes,including the maintenance and differentiation of neural stem cells(NSCs).Interestingly,while complete deletion of Fip200 severely impairs NSC maintenance and differentiation,inhibiting canonical autophagy via deletion of core genes,such as Atg5,Atg16l1,and Atg7,or blockade of canonical interactions between FIP200 and ATG13(designated as FIP200-4A mutant or FIP200 KI)does not produce comparable detrimental effects.This highlights the likely critical involvement of the non-canonical functions of FIP200,the mechanisms of which have remained elusive.Here,utilizing genetic mouse models,we demonstrated that FIP200 mediates non-canonical autophagic degradation of p62/sequestome1,primarily via TAX1BP1 in NSCs.Conditional deletion of Tax1bp1 in fip200hGFAP conditional knock-in(cKI)mice led to NSC deficiency,resembling the fip200hGFAP conditional knockout(cKO)mouse phenotype.Notably,reintroducing wild-type TAX1BP1 not only restored the maintenance of NSCs derived from tax1bp1-knockout fip200hGFAP cKI mice but also led to a marked reduction in p62 aggregate accumulation.Conversely,a TAX1BP1 mutant incapable of binding to FIP200 or NBR1/p62 failed to achieve this restoration.Furthermore,conditional deletion of Tax1bp1 in fip200hGFAP cKO mice exacerbated NSC deficiency and p62 aggregate accumulation compared to fip200hGFAP cKO mice.Collectively,these findings illustrate the essential role of the FIP200-TAX1BP1 axis in mediating the non-canonical autophagic degradation of p62 aggregates towards NSC maintenance and function,presenting novel therapeutic targets for neurodegenerative diseases.
基金This work was supported by the National Natural Science Foundation of China(2017YFA0506001,81900233,81770261,81521001,and 91749128)the Postdoctoral Science Foundation of China(2019M661375).
文摘Acute myocardial infarction(MI),one of the most common cardiovascular emergencies,is a leading cause of morbidity and mortality.Ample evidence has revealed an essential role for inflammasome activation and autophagy in the pathogenesis of acute MI.Tax1-binding protein 1(TAX1BP1),an adaptor molecule involved in termination of proinflammatory signaling,serves as an important selective autophagy adaptor,but its role in cardiac ischemia remains elusive.This study examined the role of TAX1BP1 in myocardial ischemic stress and the underlying mechanisms involved.Levels of TAX1BP1 were significantly downregulated in heart tissues of patients with ischemic heart disease and in a left anterior descending(LAD)ligation-induced model of acute MI.Adenovirus carrying TAX1BP1 was delivered into the myocardium.The acute MI induced procedure elicited an infarct and cardiac dysfunction,the effect of which was mitigated by TAX1BP1 overexpression with little effect from viral vector alone.TAX1BP1 nullified acute MI-induced activation of the NLRP3 inflammasome and associated mitochondrial dysfunction.TAX1BP1 overexpression suppressed NLRP3 mitochondrial localization by inhibiting the interaction of NLRP3 with mitochondrial antiviral signaling protein(MAVS).Further investigation revealed that ring finger protein 34(RNF34)was recruited to interact with TAX1BP1 thereby facilitating autophagic degradation of MAVS through K27-linked polyubiquitination of MAVS.Knockdown of RNF34 using siRNA nullified TAX1BP1 yielded protection against hypoxia-induced MAVS mitochondrial accumulation,NLRP3 inflammasome activation and associated loss of mitochondrial membrane potential.Taken together,our results favor a cardioprotective role for TAX1BP1 in acute MI through repression of inflammasome activation in a RNF34/MAVS-dependent manner.
基金This research was carried out within the State Assignment of the Ministry of Science and Higher Education of the Russian Federation(project FZGW-2024-0003).
文摘Objective:The objective of this study was to determine the level of methotrexate(MTX)toxicity in the intestines of mice and to evaluate the protective effect of probiotics composed of Streptococcus,Bifidobacterium,and Lactobacillus species on intestinal cells during MTX treatment.Methods:Mice were divided into three groups:control,MTX group(received MTX injections),and MTX+probiotics group(received MTX injections along with a diet containing probiotics).Morphological and histological changes,the level of mitochondrial DNA(mtDNA)damage,the level of lipid peroxidation products,and gene expression in the mice’s small intestine were assessed.Results:We demonstrated that intraperitoneal MTX injections significantly increased mtDNA damage in the liver(p<0.001),small intestine(p<0.001),and blood of mice(p<0.01).MTX elevated the quantity of lipid peroxidation products in the liver and small intestine,indicating its strong prooxidative properties.MTX induced structural changes in the mice’s intestines,characterized by leukocytic infiltration of tissues.Probiotic therapy in mice partially mitigated the morphological and histological changes in the small intestine induced by MTX,reduced oxidative stress,and promoted increased expression of quinone oxidoreductase 1(Nqo1),which participates in both cell protection against oxidative stress and drug/xenobiotic detoxification.Probiotics prevented the upregulation of the proinflammatory cytokine IL-1b in the small intestine and induced increased expression of genes associated with the Nuclear factor erythroid 2-related factor 2/Antioxidant response element(Nrf2/ARE)pathway,an important mechanism of cell protection.Conclusions:Probiotics can be considered an effective approach to reducing the toxicity of MTX during psoriasis or cancer treatment.
