基于RNA-Seq技术探讨肝失疏泄状态人群的转录组学特征
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摘要
目的 :基于RNA-Seq技术分析肝失疏泄人群的差异表达基因,从转录组水平进一步揭示"肝主疏泄"的分子机制。方法:本研究基于中医有关"肝气郁结"的表现特征,并引入现代心理学中定义为负性情绪体验的神经质人格的概念及评判标准,筛选肝失疏泄人群,通过Hiseq2000 RNA-Seq技术进行测序,分析筛选与肝主疏泄组相比,肝失疏泄人群的显著差异表达基因。然后对两组的差异基因进行GO功能的显著性富集分析以及KEGG通路的显著性富集分析。结果:肝失疏泄人群与肝主疏泄组相比有55个显著差异基因,其中上调基因有35个,下调基因有20个。GO功能显著性富集分析结果显示,这些差异表达基因与防御反应、细胞吞噬、免疫球蛋白黏附等功能密切相关。KEGG通路显著富集分析结果表明这些基因与系统性红斑狼疮疾病的发病机制关系密切。其中FCGR1C、FCGR1B、LCN2与IL-10四个差异基因差异倍数较高且上述通路上均显著富集。结论:"肝主疏泄"的内在分子机制与机体的免疫系统功能密切相关,并进一步推测其功能是通过调控上述差异基因的功能来实现的。
Objective: To analyze differentially expressed genes in patients with liver failed in controlling conveyance and dispersion based on RNA-Seq technique, and further reveal the molecular mechanism of ‘liver controlling conveyance and dispersion' at transcriptome level. Methods: Based on the characteristics of ‘stagnation of liver qi', and introducing the concept and criteria of neurotic personality defined as negative emotional experience in modern psychology, this study screened people with liver failed in controlling conveyance and dispersion, sequenced them by Hiseq2000 RNA-Seq technology, and analyzed and screened the significant differentially expressed genes in the group with liver failed in controlling conveyance and dispersion compared with the group with liver controlling conveyance and dispersion. The significant enrichment analysis of GO function and KEGG pathway were performed for the two groups of differentially expressed genes. Results: There were 55 significant differences between the group of liver failed in controlling conveyance and dispersion and the group of liver controlling conveyance and dispersion, of which 35 were up-regulated genes and 20 were down-regulated genes. Significant enrichment analysis of GO function showed that these differentially expressed genes were closely related to defense response, phagocytosis and immunoglobulin binding. Significant enrichment analysis of KEGG pathway indicated that these genes were closely related to the pathogenesis of systemic lupus erythematosus. Among them, four differentially expressed genes, FCGR1 C, FCGR1 B, LCN2 and IL-10, had higher differential multiples and were significantly enriched in the above pathways. Conclusion: The intrinsic molecular mechanism of ‘liver controlling conveyance and dispersion' is closely related to the immune system function of the organism. It is further speculated that its function is realized by regulating the functions of the above differential genes.
Objective: To analyze differentially expressed genes in patients with liver failed in controlling conveyance and dispersion based on RNA-Seq technique, and further reveal the molecular mechanism of ‘liver controlling conveyance and dispersion' at transcriptome level. Methods: Based on the characteristics of ‘stagnation of liver qi', and introducing the concept and criteria of neurotic personality defined as negative emotional experience in modern psychology, this study screened people with liver failed in controlling conveyance and dispersion, sequenced them by Hiseq2000 RNA-Seq technology, and analyzed and screened the significant differentially expressed genes in the group with liver failed in controlling conveyance and dispersion compared with the group with liver controlling conveyance and dispersion. The significant enrichment analysis of GO function and KEGG pathway were performed for the two groups of differentially expressed genes. Results: There were 55 significant differences between the group of liver failed in controlling conveyance and dispersion and the group of liver controlling conveyance and dispersion, of which 35 were up-regulated genes and 20 were down-regulated genes. Significant enrichment analysis of GO function showed that these differentially expressed genes were closely related to defense response, phagocytosis and immunoglobulin binding. Significant enrichment analysis of KEGG pathway indicated that these genes were closely related to the pathogenesis of systemic lupus erythematosus. Among them, four differentially expressed genes, FCGR1 C, FCGR1 B, LCN2 and IL-10, had higher differential multiples and were significantly enriched in the above pathways. Conclusion: The intrinsic molecular mechanism of ‘liver controlling conveyance and dispersion' is closely related to the immune system function of the organism. It is further speculated that its function is realized by regulating the functions of the above differential genes.
引文
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[3]田蕾,吴昊,韦昱,等.从心理应激理论探讨中医肝主疏泄功能的研究思考.世界中医药,2016,11(9):1905-1908
[4]岳广欣.束缚应激大鼠中枢AMPA受体及相关蛋白变化与肝主疏泄的关系.北京:北京中医药大学:2006
[5]李聪,谢鸣,赵荣华,等.柴疏四君汤对肝郁、脾虚和肝郁脾虚模型大鼠性腺轴的影响.中国中西医结合杂志,2014,34(6):694-697
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[8]贾昌路,张瑶,朱玲,等.转录组测序技术在生物测序中的应用研究进展.分子植物育种,2015,13(10):2388-2394
[9]杜智慧.基于RNA-Seq筛选抗绵羊肺炎支原体差异表达基因.石河子:石河子大学,2018
[10]唐利龙,邓华亮,赵勇,等.健康大学生生活事件与躯体健康状况的关系研究.世界中医药,2013,8(2):117-121
[11]吴梦莹,周仁来,黄雅梅,等.神经质程度和月经周期对女性主观情绪和生理反应的影响.心理学报,2014,46(1):58-68
[12]朱文峰.中医诊断学.北京:中国中医药出版社,2007:186
[13]宋萍,詹向红,杨丽萍,等.五脏与抑郁症发病探析.中华中医药杂志,2013,28(10):2851-2854
[14]王华民,徐军发,陈雪玲.临床免疫学.北京:军事医学科学出版社,2010:6-7
[15]刘仕琦,贾宇平,张峰,等.从摘除松果腺大鼠与肝郁大鼠MT、5-HT、NO和免疫功能的变化探讨“肝应春”的作用机制.中华中医药杂志,2016,31(10):4194-4197
[16]严灿,张斩春,邓中炎.肝主疏泄免疫学机制的临床与实验研究.中国中医基础医学杂志,1995,1(3):36-38
[17]刘建鸿,岳嘉,段永强,等.肝郁脾虚证模型大鼠免疫细胞分化和增殖变化规律研究.中国中医药信息杂志,2013,20(1):43-45
[18]李雪.系统性红斑狼疮患者外周血NK细胞抑制性受体TIGIT表达和意义.南昌:南昌大学,2018
[19]吴海竞,陆前进.系统性红斑狼疮发病机制的研究进展.皮肤科学通报,2018,35(3):249-257,235
[20]Berger T,Togawa A,Duncan G S,et al.Lipocalin 2-deficient mice exhibit increased sensitivity to Escherichia coli infection but not to ischemia-reperfusion injury.Proceedings of the National Academy of Sciences of the United States of America,2006,103(6):1834-1839
[21]Maresco D L,Chang E,Theil K S,et al.The three genes of the human FCGR1 gene family encoding Fc gamma RI flank the centromere of chromosome 1 at 1p12 and 1q21.Cytogenetics and Cell Genetics,1996,73(3):157-163
[2]李德新.中医基础理论讲稿.北京:中国医药科技出版社,2006:127-132
[3]田蕾,吴昊,韦昱,等.从心理应激理论探讨中医肝主疏泄功能的研究思考.世界中医药,2016,11(9):1905-1908
[4]岳广欣.束缚应激大鼠中枢AMPA受体及相关蛋白变化与肝主疏泄的关系.北京:北京中医药大学:2006
[5]李聪,谢鸣,赵荣华,等.柴疏四君汤对肝郁、脾虚和肝郁脾虚模型大鼠性腺轴的影响.中国中西医结合杂志,2014,34(6):694-697
[6]朱静,夏天,刘兆娟.肝郁-心理应激对子宫肌瘤大鼠性激素及免疫因子影响的实验研究.辽宁中医药大学学报,2011,13(9):52-56
[7]Cosla V,Angelini C,DeFeis I,CiccodicolaA.Uncoveringthe complexity of transcriptomes with RNA-seq.Biomed Biotechnol,2010,85:3916
[8]贾昌路,张瑶,朱玲,等.转录组测序技术在生物测序中的应用研究进展.分子植物育种,2015,13(10):2388-2394
[9]杜智慧.基于RNA-Seq筛选抗绵羊肺炎支原体差异表达基因.石河子:石河子大学,2018
[10]唐利龙,邓华亮,赵勇,等.健康大学生生活事件与躯体健康状况的关系研究.世界中医药,2013,8(2):117-121
[11]吴梦莹,周仁来,黄雅梅,等.神经质程度和月经周期对女性主观情绪和生理反应的影响.心理学报,2014,46(1):58-68
[12]朱文峰.中医诊断学.北京:中国中医药出版社,2007:186
[13]宋萍,詹向红,杨丽萍,等.五脏与抑郁症发病探析.中华中医药杂志,2013,28(10):2851-2854
[14]王华民,徐军发,陈雪玲.临床免疫学.北京:军事医学科学出版社,2010:6-7
[15]刘仕琦,贾宇平,张峰,等.从摘除松果腺大鼠与肝郁大鼠MT、5-HT、NO和免疫功能的变化探讨“肝应春”的作用机制.中华中医药杂志,2016,31(10):4194-4197
[16]严灿,张斩春,邓中炎.肝主疏泄免疫学机制的临床与实验研究.中国中医基础医学杂志,1995,1(3):36-38
[17]刘建鸿,岳嘉,段永强,等.肝郁脾虚证模型大鼠免疫细胞分化和增殖变化规律研究.中国中医药信息杂志,2013,20(1):43-45
[18]李雪.系统性红斑狼疮患者外周血NK细胞抑制性受体TIGIT表达和意义.南昌:南昌大学,2018
[19]吴海竞,陆前进.系统性红斑狼疮发病机制的研究进展.皮肤科学通报,2018,35(3):249-257,235
[20]Berger T,Togawa A,Duncan G S,et al.Lipocalin 2-deficient mice exhibit increased sensitivity to Escherichia coli infection but not to ischemia-reperfusion injury.Proceedings of the National Academy of Sciences of the United States of America,2006,103(6):1834-1839
[21]Maresco D L,Chang E,Theil K S,et al.The three genes of the human FCGR1 gene family encoding Fc gamma RI flank the centromere of chromosome 1 at 1p12 and 1q21.Cytogenetics and Cell Genetics,1996,73(3):157-163