结核肺组织与正常肺组织蛋白质组双向电泳图谱的差异分析
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摘要
目的对结核肺组织与正常肺组织双向电泳图谱进行比较分析,筛选差异表达蛋白质。方法利用双向电泳分离结核组及正常组总蛋白质,并通过计算机图像分析软件分析电泳图谱。结果在结核肺组织中检测有效蛋白点数为(190±11)个,正常肺组织中检测有效蛋白点数为(179±5)个。发现两者间共有15个差异蛋白点,蛋白点分子量(Mr)主要分布于15kD～80kD,PI为3～10。4个点在病灶组织中表达上调,11个点在病灶组织中表达下调。结论通过双向凝胶电泳和图像分析技术对结核肺组织及正常肺组织蛋白质组进行研究,获得了结核组与正常组的差异蛋白点,为进一步利用质谱分析技术做差异蛋白质鉴定奠定了基础,并为肺结核的早期诊断、发病机制探索、病情监测提供依据。
Objective To analyze the two dimentional electrophoresis( 2-DE) maps of lung tissue from pulmonary tuberculosis and normal lesions and screen differently expressed proteins between them. Methods 2-DE( two dimentional electrophoresis) was used to sever the general protein from lung tissues in pulmonary tuberculosis and normal lesions,then the results were analyzed by computer-graph analysis software. Results 190 ± 11 protein spots were verified in pulmonary tuberculosis tissue and 179 ± 5 ones in normal lung tissue. There were 15 discrepancy protein spots. The protein point molecular weight( Mr) is mainly distributed in 15 kD ～ 80 kD,isoelectric point( PI) is 3 ～ 10. Meanwhile,4 spots frequently emerged and 11 ones were less expressed in pulmonary tuberculosis tissue. Conclusion The discrepancy protein spots were identified,which laid a solid foundation for further research of discrepancy protein,and provide basis for early diagnosis,pathogenesis and monitoring of pulmonary tuberculosis.
Objective To analyze the two dimentional electrophoresis( 2-DE) maps of lung tissue from pulmonary tuberculosis and normal lesions and screen differently expressed proteins between them. Methods 2-DE( two dimentional electrophoresis) was used to sever the general protein from lung tissues in pulmonary tuberculosis and normal lesions,then the results were analyzed by computer-graph analysis software. Results 190 ± 11 protein spots were verified in pulmonary tuberculosis tissue and 179 ± 5 ones in normal lung tissue. There were 15 discrepancy protein spots. The protein point molecular weight( Mr) is mainly distributed in 15 kD ～ 80 kD,isoelectric point( PI) is 3 ～ 10. Meanwhile,4 spots frequently emerged and 11 ones were less expressed in pulmonary tuberculosis tissue. Conclusion The discrepancy protein spots were identified,which laid a solid foundation for further research of discrepancy protein,and provide basis for early diagnosis,pathogenesis and monitoring of pulmonary tuberculosis.
引文
[1]杨柳.巨噬细胞在结核性肉芽肿形成和发展过程中的极化[D].江西:南昌大学,2013.
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[6]黄迪希,刘志辉,杨瑜,等.结核性胸腔积液的双向电泳蛋白质组学[J].实用医学杂志,2016,32(14):2326-2329.
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[8]李昕洁,谭守勇,刘玉美,等.结核病患者血清蛋白质/多肽谱的初步研究[J].广东医学,2010,31(15):1928-1930.
[9]李文胜.肺结核病灶组织与正常组织基因表达谱的差异和部分肺外结核临床分离菌株基因分型的特点[D].北京:北京市结核病胸部肿瘤研究所,2011.
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[11]侯江厚,程小星,张灵霞,等.结核病局部免疫的研究进展[J].中华结核和呼吸杂志,2014,37(2):119-121.
[12]彭卫生,王英年,肖成志.新编结核病学[M].2版,北京:中国医药科技出版社,2003:123-124.
[13]BARK C M,MANCEUR A M,MALONE L S,et al.Identification of Host Proteins Predictive of Early Stage Mycobacterium tuberculosis Infection[J].EbioMedicine,2017,21:150-157.
[14]BANAEI-ESFAHANI A,NICOD C,AEBERSOLD R,et al.Systems proteomics approaches to study bacterial pathogens:application to Mycobacterium tuberculosis[J].Curr Opin Microbiol,2017,39:64-72.
[15]劳穗华,谢贝,董海平等.肺结核治疗过程中血清IL-10、IL-12p40、IL-12p70、IL-17A、IL-4、IL-6和IPl0水平动态观察[J].现代医院,2016,16(8):1172-1174.
[2]PIETERS J.Mycobacterium tuberculosis and the macrophage:maintaining a Balance[J].Cell Host Microbe,2008,3(6):399-407.
[3]张丽霞,冯爽,张立.蛋白质组学在结核病诊断中的研究进展[J].现代预防医学,2014,41(17):3206-3207,3210.
[4]LIST E O,BERRYMAN D E,BOWER B,et al.The Use of Proteomics to Study Infectious Diseases[J],Infect Disord Drug Targets,2008,8(1):31-45.
[5]李泰明,杨幸远,蒙青林,等.表达结核分枝杆菌热休克蛋白16.3的U937细胞蛋白组学分析[J].中华结核和呼吸杂志,2015,38(1):34-38.
[6]黄迪希,刘志辉,杨瑜,等.结核性胸腔积液的双向电泳蛋白质组学[J].实用医学杂志,2016,32(14):2326-2329.
[7]张树明,白海玉,王伟明,等.肺部疾病的蛋白质组学研究[J].黑龙江中医药,2012,4(41):43-44.
[8]李昕洁,谭守勇,刘玉美,等.结核病患者血清蛋白质/多肽谱的初步研究[J].广东医学,2010,31(15):1928-1930.
[9]李文胜.肺结核病灶组织与正常组织基因表达谱的差异和部分肺外结核临床分离菌株基因分型的特点[D].北京:北京市结核病胸部肿瘤研究所,2011.
[10]RYAN G J,HOFF D R,DRIVER E R,et al.Multiple M.tuberculosis Phenotypes in Mouse and Guinea Pig Lung Tissue Revealed by a Dual-Staining Approach[J].PLo S ONE,2010,5(6):e11108.
[11]侯江厚,程小星,张灵霞,等.结核病局部免疫的研究进展[J].中华结核和呼吸杂志,2014,37(2):119-121.
[12]彭卫生,王英年,肖成志.新编结核病学[M].2版,北京:中国医药科技出版社,2003:123-124.
[13]BARK C M,MANCEUR A M,MALONE L S,et al.Identification of Host Proteins Predictive of Early Stage Mycobacterium tuberculosis Infection[J].EbioMedicine,2017,21:150-157.
[14]BANAEI-ESFAHANI A,NICOD C,AEBERSOLD R,et al.Systems proteomics approaches to study bacterial pathogens:application to Mycobacterium tuberculosis[J].Curr Opin Microbiol,2017,39:64-72.
[15]劳穗华,谢贝,董海平等.肺结核治疗过程中血清IL-10、IL-12p40、IL-12p70、IL-17A、IL-4、IL-6和IPl0水平动态观察[J].现代医院,2016,16(8):1172-1174.