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脂多糖刺激后释放的单核细胞微囊泡(微粒)的红外光谱表征

Infrared spectroscopic characterization of monocytic microvesicles (microparticles) released upon lipopolysaccharide stimulation.

作者信息

Lee Joonsup, Wen Beryl, Carter Elizabeth A, Combes Valery, Grau Georges E R, Lay Peter A

机构信息

School of Chemistry and Vibrational Spectroscopy Core Facility, The University of Sydney, Sydney, New South Wales, Australia.

Vascular Immunopathology Unit, Bosch Institute-School of Medical Sciences, and.

出版信息

FASEB J. 2017 Jul;31(7):2817-2827. doi: 10.1096/fj.201601272R. Epub 2017 Mar 17.

Abstract

Microvesicles (MVs) are involved in cell-cell interactions, including disease pathogenesis. Nondestructive Fourier-transform infrared (FTIR) spectra from MVs were assessed as a technique to provide new biochemical insights into a LPS-induced monocyte model of septic shock. FTIR spectroscopy provided a quick method to investigate relative differences in biomolecular content of different MV populations that was complementary to traditional semiquantitative omics approaches, with which it is difficult to provide information on relative changes between classes (proteins, lipids, nucleic acids, carbohydrates) or protein conformations. Time-dependent changes were detected in biomolecular contents of MVs and in the monocytes from which they were released. Differences in phosphatidylcholine and phosphatidylserine contents were observed in MVs released under stimulation, and higher relative concentrations of RNA and α-helical structured proteins were present in stimulated MVs compared with MVs from resting cells. FTIR spectra of stimulated monocytes displayed changes that were consistent with those observed in the corresponding MVs they released. LPS-stimulated monocytes had reduced concentrations of nucleic acids, α-helical structured proteins, and phosphatidylcholine compared with resting monocytes but had an increase in total lipids. FTIR spectra of MV biomolecular content will be important in shedding new light on the mechanisms of MVs and the different roles they play in physiology and disease pathogenesis.-Lee, J., Wen, B., Carter, E. A., Combes, V., Grau, G. E. R., Lay, P. A. Infrared spectroscopic characterization of monocytic microvesicles (microparticles) released upon lipopolysaccharide stimulation.

摘要

微泡(MVs)参与细胞间相互作用,包括疾病发病机制。对MVs的无损傅里叶变换红外(FTIR)光谱进行了评估,作为一种为脂多糖诱导的脓毒症休克单核细胞模型提供新的生化见解的技术。FTIR光谱提供了一种快速方法,用于研究不同MV群体生物分子含量的相对差异,这是对传统半定量组学方法的补充,传统方法难以提供关于不同类别(蛋白质、脂质、核酸、碳水化合物)之间相对变化或蛋白质构象的信息。检测到MVs及其释放来源的单核细胞生物分子含量随时间的变化。在刺激条件下释放的MVs中观察到磷脂酰胆碱和磷脂酰丝氨酸含量的差异,与静息细胞来源的MVs相比,刺激后释放的MVs中RNA和α-螺旋结构蛋白的相对浓度更高。刺激后单核细胞的FTIR光谱显示出的变化与它们释放的相应MVs中观察到的变化一致。与静息单核细胞相比,脂多糖刺激的单核细胞中核酸、α-螺旋结构蛋白和磷脂酰胆碱的浓度降低,但总脂质增加。MV生物分子含量的FTIR光谱对于揭示MVs的机制及其在生理和疾病发病机制中所起的不同作用具有重要意义。-李,J.,温,B.,卡特,E.A.,孔贝斯,V.,格劳,G.E.R.,莱,P.A.脂多糖刺激后单核细胞微泡(微粒)的红外光谱表征

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