Background:Based on previous theoretical studies,JQ-1 as a common inhibitor of bromodomain and extraterminal(BET)proteins was used to treat a variety of diseases.Therefore,we aimed to explore the mechanism of action o...Background:Based on previous theoretical studies,JQ-1 as a common inhibitor of bromodomain and extraterminal(BET)proteins was used to treat a variety of diseases.Therefore,we aimed to explore the mechanism of action of JQ-1 on BET proteins based on bioinformatics and build the novel hypothesis of JQ-1 in treating atherosclerosis(AS)caused by proliferation of vascular smooth muscle cells(VSMCs).Methods:We selected the chip GSE138323 which was searched with the key words“Vascular smooth muscle cell proliferation”in Gene Expression Omnibus(GEO)database,and differential gene analysis was performed between the GRO and JQ-1 groups.Then the top twenty significantly up-regulated genes and the top twenty significantly down-regulated genes were selected for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.Thirdly,structured the PPI network of forty differential genes,and the core genes were screened by using the MCC algorithm which in“Cytohubba”plugin in the Cytoscapev3.9.1 software.After that,single gene Gene Set Enrichment Analysis(GSEA)enrichment analysis was performed on the selected core genes in R language.Finally molecular docking validation was performed.Results:Five core genes was selected:H3C2,H3C4,H3C7,H3C10 and AREG.The GO enrichment analysis results showed that there were twenty-five entries in biological process,eight entries in cellular components(CC),and twenty-five entries in molecular function.The KEGG enrichment analysis results showed that there were seven pathways,mainly including systemic lupus erythematosus and external neutrophil trap formation.The GSEA results showed that the five genes were mainly through the regulation of cytochrome P450 metabolism,PPAR signaling pathway and other pathways.The molecular docking results showed that JQ-1 had binding activity with these five genes.Conclusions:JQ-1 may regulate the expression of the genes that H3C2,H3C4,H3C7,H3C10 and AREG,to mainly regulate the genes in cytochrome P450 metabolism,PPAR singling pathway and other pathways,to make some influence in the proliferation of VSMCs,and improved atherosclerotic symptoms due to vascular smooth muscle proliferation,thus treating cardiovascular disease.展开更多
Subject Code:H16With the support by the National Natural Science Foundation of China,a collaborative study by the research groups led by Prof.Wang Chenji(王陈继)from Fudan University,Prof.Huang Haojie(黄浩杰)from Mayo...Subject Code:H16With the support by the National Natural Science Foundation of China,a collaborative study by the research groups led by Prof.Wang Chenji(王陈继)from Fudan University,Prof.Huang Haojie(黄浩杰)from Mayo Clinic and Sun Yinhao(孙颖浩)from the Second Military Medical University have展开更多
溴结构域和超末端结构域(bromodomain and extra-terminal,BET)蛋白家族作为表观"阅读器",在哺乳动物发育过程中起着至关重要的作用。其家族内的各成员通过识别各种表观修饰并募集相应的功能复合物,对相关基因进行精密调控,...溴结构域和超末端结构域(bromodomain and extra-terminal,BET)蛋白家族作为表观"阅读器",在哺乳动物发育过程中起着至关重要的作用。其家族内的各成员通过识别各种表观修饰并募集相应的功能复合物,对相关基因进行精密调控,促使早期胚胎向特定方向分化和发育。另外,随着诱导性多潜能干细胞(induced pluripotent stem cell,iPSC)重编程技术发展,越来越多的研究发现BET蛋白家族在体细胞重编程中可能也占据着核心地位。本文总结了BET蛋白家族在哺乳动物发育和iPSC重编程中的作用,并对BET家族调控重编程的新机制进行了展望。展开更多
旨在探究抑制水牛BET(bromodomain and extra terminal,BET)蛋白对支持细胞自噬调控的影响。以水牛支持细胞(sertoli cells,SCs)为研究对象,首先从睾丸中分离纯化得到高纯度的SCs,通过免疫荧光法及RT-PCR法检测出水牛SCs的标记基因均正...旨在探究抑制水牛BET(bromodomain and extra terminal,BET)蛋白对支持细胞自噬调控的影响。以水牛支持细胞(sertoli cells,SCs)为研究对象,首先从睾丸中分离纯化得到高纯度的SCs,通过免疫荧光法及RT-PCR法检测出水牛SCs的标记基因均正常表达。结果显示,BET蛋白被抑制后,水牛SCs形态和数量发生显著变化,细胞的体积变大,数量减少,折光性减弱,免疫荧光分析和MDC分析也证实其生物学功能受到影响。超微结构分析发现,水牛SCs中产生大量自噬体、自噬溶酶体以及自噬空泡,且自噬相关标记基因的表达量显著提升。说明抑制BET蛋白可引起水牛SCs活性和自噬活性降低,并导致自噬体增多。结果表明,本试验为BET蛋白调控雄性生殖机制的研究提供了依据。展开更多
目的:探讨溴结构域(bromodomain and extra-terminal,BET)蛋白抑制剂JQ1对系统性红斑狼疮(systemic lupus erythematosus,SLE)外周血CD4^+T细胞中自身免疫相关基因表达的作用。方法:利用磁珠分选获得SLE患者CD4^+T细胞,采用流式细胞检测...目的:探讨溴结构域(bromodomain and extra-terminal,BET)蛋白抑制剂JQ1对系统性红斑狼疮(systemic lupus erythematosus,SLE)外周血CD4^+T细胞中自身免疫相关基因表达的作用。方法:利用磁珠分选获得SLE患者CD4^+T细胞,采用流式细胞检测CD4^+T细胞纯度。用JQ1(100 nm/L)处理CD4^+T细胞6,24,48 h后收集细胞。采用实时荧光定量PCR(quantitative real-time PCR,q PCR)检测不同辅助性T细胞亚类特异性基因的m RNA表达。采用ELISA法检测细胞培养上清中细胞因子的蛋白水平。结果:磁珠分选所得CD4^+T细胞百分比为97.2%。与对照组相比,JQ1处理组IFNG,IL-17F,IL-21,CXCR5和FOXP3 m RNA表达水平在6,24,48 h均显著降低(P<0.05),IL-17A m RNA表达在6,24 h显著下降(P<0.01),IL-4 m RNA表达在24,48 h显著升高(P<0.01),TGF-β1 m RNA表达在6,48 h显著升高(P<0.05)。JQ1处理48 h细胞培养上清中IFN-γ,IL-21和IL-17蛋白水平明显下降(P<0.05),IL-4和TGF-β蛋白水平明显升高(P<0.05)。结论:BET蛋白抑制剂JQ1可纠正SLE患者CD4^+T细胞中的免疫调节失衡,促进免疫稳态恢复。展开更多
为了明确鸡(Gallus gallus)溴结构域和超末端结构(bromodomain and extra terminal,BET)家族蛋白亚细胞定位及其基因在免疫器官中的表达特性,并为后续研究BET家族蛋白调控鸡相关病毒复制的作用机制提供基础,本研究将前期构建的鸡BRD2、B...为了明确鸡(Gallus gallus)溴结构域和超末端结构(bromodomain and extra terminal,BET)家族蛋白亚细胞定位及其基因在免疫器官中的表达特性,并为后续研究BET家族蛋白调控鸡相关病毒复制的作用机制提供基础,本研究将前期构建的鸡BRD2、BRD3、BRD4基因重组真核表达载体pEGFP-BRD2、pEGFP-BRD3和pEGFP-BRD4分别转染人(Homo sapiens)胚胎肾细胞(HEK-293T),利用Western blotting和荧光检测鸡BRD2、BRD3、BRD4基因的表达和蛋白的亚细胞定位情况,并利用实时定量PCR(real time quantitative PCR)技术检测1~6月龄鸡免疫器官中BRD2、BRD3和BRD4基因的表达。结果表明,鸡BET家族蛋白融合蛋白EGFP-BRD2、EGFP-BRD3和EGFP-BRD4在细胞中正确表达,且鸡BRD2、BRD3和BRD4蛋白主要定位在细胞核。实时定量PCR检测结果发现,鸡的脾脏和胸腺中BRD2基因的表达量在3月龄时最高,在法氏囊中则在4月龄时最高,表达变化均为“倒V”型;脾脏中BRD3基因的表达量在1月龄时为最高;而鸡的胸腺和法氏囊中,BRD3和BRD4基因的表达量均在6月龄时为最高,其中,BRD3基因在胸腺和法氏囊的表达变化,以及BRD4基因在法氏囊中的表达变化均为逐渐上升的“直线”型;BRD3基因在脾脏中的表达变化,以及BRD4基因在胸腺中的表达变化均为“倒N”型;脾脏中BRD4基因的表达量在3月龄时最高,其表达变化呈先上升后下降的趋势。鸡BET家族基因在不同月龄鸡的脾脏、胸腺和法氏囊中均有表达,但表达量呈现不同的变化趋势,BET家族蛋白主要定位在细胞核。展开更多
基金supported by a grant from Key Project of Education Commission of Hubei Province(D20202802)Hubei Key Laboratory of Diabetes and Angiopathy Program(2020XZ10)of Hubei University of Science.
文摘Background:Based on previous theoretical studies,JQ-1 as a common inhibitor of bromodomain and extraterminal(BET)proteins was used to treat a variety of diseases.Therefore,we aimed to explore the mechanism of action of JQ-1 on BET proteins based on bioinformatics and build the novel hypothesis of JQ-1 in treating atherosclerosis(AS)caused by proliferation of vascular smooth muscle cells(VSMCs).Methods:We selected the chip GSE138323 which was searched with the key words“Vascular smooth muscle cell proliferation”in Gene Expression Omnibus(GEO)database,and differential gene analysis was performed between the GRO and JQ-1 groups.Then the top twenty significantly up-regulated genes and the top twenty significantly down-regulated genes were selected for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.Thirdly,structured the PPI network of forty differential genes,and the core genes were screened by using the MCC algorithm which in“Cytohubba”plugin in the Cytoscapev3.9.1 software.After that,single gene Gene Set Enrichment Analysis(GSEA)enrichment analysis was performed on the selected core genes in R language.Finally molecular docking validation was performed.Results:Five core genes was selected:H3C2,H3C4,H3C7,H3C10 and AREG.The GO enrichment analysis results showed that there were twenty-five entries in biological process,eight entries in cellular components(CC),and twenty-five entries in molecular function.The KEGG enrichment analysis results showed that there were seven pathways,mainly including systemic lupus erythematosus and external neutrophil trap formation.The GSEA results showed that the five genes were mainly through the regulation of cytochrome P450 metabolism,PPAR signaling pathway and other pathways.The molecular docking results showed that JQ-1 had binding activity with these five genes.Conclusions:JQ-1 may regulate the expression of the genes that H3C2,H3C4,H3C7,H3C10 and AREG,to mainly regulate the genes in cytochrome P450 metabolism,PPAR singling pathway and other pathways,to make some influence in the proliferation of VSMCs,and improved atherosclerotic symptoms due to vascular smooth muscle proliferation,thus treating cardiovascular disease.
文摘Subject Code:H16With the support by the National Natural Science Foundation of China,a collaborative study by the research groups led by Prof.Wang Chenji(王陈继)from Fudan University,Prof.Huang Haojie(黄浩杰)from Mayo Clinic and Sun Yinhao(孙颖浩)from the Second Military Medical University have
文摘旨在探究抑制水牛BET(bromodomain and extra terminal,BET)蛋白对支持细胞自噬调控的影响。以水牛支持细胞(sertoli cells,SCs)为研究对象,首先从睾丸中分离纯化得到高纯度的SCs,通过免疫荧光法及RT-PCR法检测出水牛SCs的标记基因均正常表达。结果显示,BET蛋白被抑制后,水牛SCs形态和数量发生显著变化,细胞的体积变大,数量减少,折光性减弱,免疫荧光分析和MDC分析也证实其生物学功能受到影响。超微结构分析发现,水牛SCs中产生大量自噬体、自噬溶酶体以及自噬空泡,且自噬相关标记基因的表达量显著提升。说明抑制BET蛋白可引起水牛SCs活性和自噬活性降低,并导致自噬体增多。结果表明,本试验为BET蛋白调控雄性生殖机制的研究提供了依据。
文摘目的:探讨溴结构域(bromodomain and extra-terminal,BET)蛋白抑制剂JQ1对系统性红斑狼疮(systemic lupus erythematosus,SLE)外周血CD4^+T细胞中自身免疫相关基因表达的作用。方法:利用磁珠分选获得SLE患者CD4^+T细胞,采用流式细胞检测CD4^+T细胞纯度。用JQ1(100 nm/L)处理CD4^+T细胞6,24,48 h后收集细胞。采用实时荧光定量PCR(quantitative real-time PCR,q PCR)检测不同辅助性T细胞亚类特异性基因的m RNA表达。采用ELISA法检测细胞培养上清中细胞因子的蛋白水平。结果:磁珠分选所得CD4^+T细胞百分比为97.2%。与对照组相比,JQ1处理组IFNG,IL-17F,IL-21,CXCR5和FOXP3 m RNA表达水平在6,24,48 h均显著降低(P<0.05),IL-17A m RNA表达在6,24 h显著下降(P<0.01),IL-4 m RNA表达在24,48 h显著升高(P<0.01),TGF-β1 m RNA表达在6,48 h显著升高(P<0.05)。JQ1处理48 h细胞培养上清中IFN-γ,IL-21和IL-17蛋白水平明显下降(P<0.05),IL-4和TGF-β蛋白水平明显升高(P<0.05)。结论:BET蛋白抑制剂JQ1可纠正SLE患者CD4^+T细胞中的免疫调节失衡,促进免疫稳态恢复。
文摘为了明确鸡(Gallus gallus)溴结构域和超末端结构(bromodomain and extra terminal,BET)家族蛋白亚细胞定位及其基因在免疫器官中的表达特性,并为后续研究BET家族蛋白调控鸡相关病毒复制的作用机制提供基础,本研究将前期构建的鸡BRD2、BRD3、BRD4基因重组真核表达载体pEGFP-BRD2、pEGFP-BRD3和pEGFP-BRD4分别转染人(Homo sapiens)胚胎肾细胞(HEK-293T),利用Western blotting和荧光检测鸡BRD2、BRD3、BRD4基因的表达和蛋白的亚细胞定位情况,并利用实时定量PCR(real time quantitative PCR)技术检测1~6月龄鸡免疫器官中BRD2、BRD3和BRD4基因的表达。结果表明,鸡BET家族蛋白融合蛋白EGFP-BRD2、EGFP-BRD3和EGFP-BRD4在细胞中正确表达,且鸡BRD2、BRD3和BRD4蛋白主要定位在细胞核。实时定量PCR检测结果发现,鸡的脾脏和胸腺中BRD2基因的表达量在3月龄时最高,在法氏囊中则在4月龄时最高,表达变化均为“倒V”型;脾脏中BRD3基因的表达量在1月龄时为最高;而鸡的胸腺和法氏囊中,BRD3和BRD4基因的表达量均在6月龄时为最高,其中,BRD3基因在胸腺和法氏囊的表达变化,以及BRD4基因在法氏囊中的表达变化均为逐渐上升的“直线”型;BRD3基因在脾脏中的表达变化,以及BRD4基因在胸腺中的表达变化均为“倒N”型;脾脏中BRD4基因的表达量在3月龄时最高,其表达变化呈先上升后下降的趋势。鸡BET家族基因在不同月龄鸡的脾脏、胸腺和法氏囊中均有表达,但表达量呈现不同的变化趋势,BET家族蛋白主要定位在细胞核。
