血清外泌体的蛋白质组分析及其在骨质疏松中的应用
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摘要
外泌体是细胞分泌的微小具膜囊泡,作为重要媒介参与细胞间的信号传递,已在疾病的诊断和治疗中发挥独特作用。骨质疏松症是全身性骨代谢疾病,容易引起骨密度减少并导致骨折,在老年人群中发病率很高,目前急需发展特异性体液诊断技术。本论文采用超速离心的方法,对血清中的外泌体进行了分离富集和表征,并采用液相色谱-质谱进行了外泌体蛋白质组学分析,共鉴定到了179个外泌体蛋白质,主要参与防御响应和免疫应答等生物过程。针对来自正常对照组、骨量减少组和骨质疏松组的血清样本,分离富集其中的外泌体,通过免标记定量蛋白质组学分析,分别鉴定到188、224和185个蛋白质。定量分析显示17个蛋白质的表达量在骨质疏松组和骨量减少组有显著改变(p<0.05),包括Integrinβ3、Integrinα2β1、Talin 1和Gelsolin等,说明人体骨质在衰变过程中发生了系统性变化,并体现在血清外泌体中。该研究可为骨质疏松研究提供潜在的分子标志物,有助于阐明其病变机制。
Exosomes are tiny vesicles secreted by cells, can be important mediators of cell-to-cell communication, and play unique roles in disease diagnosis and treatment. Osteoporosis is a metabolic bone disease with high incidence in the elderly, characterized by low bone mineral density and deterioration of bone microstructure. Highly specific diagnostic methods capable of identifying early-stage osteoporosis are urgently needed. In this study, serum exosomes were comprehensively enriched and characterized. In total, 179 exosomal proteins were identified using liquid chromatography-mass spectrometry, most of which are involved in important biological processes such as defense and immune responses. Through label free quantification of serum exosomes, 188, 224, and 185 proteins were identified in the normal, osteopenia, and osteoporosis groups, respectively. Quantitative results also showed that 17 proteins were significantly(p <0.05) dysregulated in the osteoporosis and osteopenia groups, including Integrin β3, Integrin α2β1, Talin 1, and Gelsolin. This study provides potential molecular markers for osteoporosis studs and will contributes to the elucidating the pathogenesis of osteoporosis.
Exosomes are tiny vesicles secreted by cells, can be important mediators of cell-to-cell communication, and play unique roles in disease diagnosis and treatment. Osteoporosis is a metabolic bone disease with high incidence in the elderly, characterized by low bone mineral density and deterioration of bone microstructure. Highly specific diagnostic methods capable of identifying early-stage osteoporosis are urgently needed. In this study, serum exosomes were comprehensively enriched and characterized. In total, 179 exosomal proteins were identified using liquid chromatography-mass spectrometry, most of which are involved in important biological processes such as defense and immune responses. Through label free quantification of serum exosomes, 188, 224, and 185 proteins were identified in the normal, osteopenia, and osteoporosis groups, respectively. Quantitative results also showed that 17 proteins were significantly(p <0.05) dysregulated in the osteoporosis and osteopenia groups, including Integrin β3, Integrin α2β1, Talin 1, and Gelsolin. This study provides potential molecular markers for osteoporosis studs and will contributes to the elucidating the pathogenesis of osteoporosis.
引文
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[9] Khosla S,Melton L J 3rd,Riggs B L.J Bone Miner Res,2011,26(3):441
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[11] Aasis U,Gladnick B P,Eve D,et al.J Bone Joint Surg Am,2010,92(3):743
[12] Kanis J A,Melton Iii L J,Christiansen C,et al.J Bone Miner Res,1994,9(8):1137
[13] Alexandra H.JAMA,2002,25(5):164
[14] Wheater G,Elshahaly M,Tuck S P,et al.J Trans Med,2013,11:201
[15] Chen I-H,Xue L,Hsu C-C,et al.Proc Natl Acad Sci USA,2017,114(12):3175
[16] Sun Y,Huo C,Qiao Z,et al.J Proteome Res,2018,17(3):1101
[17] Buschow S I,Liefhebber J M P,Wubbolts R,et al.Blood Cells Mol Dis,2005,35(3):398
[18] Nikolic I,Stankovic N D,Bicker F,et al.Blood,2013,121(15):3041
[19] Lee D-Y,Yeh C-R,Chang S-F,et al.J Bone Miner Res,2008,23(7):1140
[20] Xu H,Wu F,Zhang H,et al.Biochimie,2017,138:184
[21] Tofteng C L,Bach-Mortensen P,Bojesen S E,et al.Pharmacogenet.Genomics,2007,17(1):85
[22] Tucci M,De Palma R,Lombardi L,et al.Cancer Res,2009,69(16):6738
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[24] Zou W,Izawa T,Zhu T,et al.Mol Cell Biol,2013,33(4):830
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