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[利用荧光光谱法研究血浆蛋白聚糖与低密度脂蛋白之间的相互作用]

[Study of the interaction between plasma proteoglycans and LDL by means of fluorescence spectroscopy].

作者信息

Siddi F, Senes A, Coinu R, Formato M, Cherchi G M

机构信息

Istituto di Biologia Applicata, Università di Sassari.

出版信息

Boll Soc Ital Biol Sper. 1992 Nov;68(11):655-61.

PMID:1297360
Abstract

The relevance of the interaction between LDL and PGs in the development of atherosclerotic processes is well known. However, the exact nature of the interaction and the consequent structural and/or conformational modifications of the lipoprotein remain to be clarified. It has been demonstrated that after this interaction the LDL particle is not recognized by specific cellular receptors and enters the scavenger pathway operating in different cell types. These effects have been shown by using aortic PGs, but PGs are also present in the plasma compartment and may interact constantly with LDL, taking part in the regulation of lipid metabolism. In order to assess the capability of plasma PGs to induce LDL modifications, we investigated their interactions by studying the changes in the organizational parameters of LDL by fluorescence spectroscopy. Plasma PGs were isolated by DEAE Sephacel ion exchange chromatography and Sephacryl S300 gel filtration in two different families: a low-charge PG and a high-charge PG. Human LDL was prepared from plasma of normolipemic donors by ultracentrifugal flotation between 1.025-1.045 g/ml. Steady-state anisotropy measures were obtained by analyzing the rotational diffusion rate of DPH after incubation of LDL with plasma PGs in a physiological ratio. In our experimental conditions, LDL incubation with plasma low-charge PG did not modify DPH fluorescence anisotropy, whereas LDL treatment with highly charged PGs induced a marked decrease of this parameter, suggesting a significant effect on LDL microviscosity. The data show that both the charge and the GAG composition of PGs appear to be critical factors in LDL-PG interaction.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

低密度脂蛋白(LDL)与蛋白聚糖(PGs)之间的相互作用在动脉粥样硬化进程发展中的相关性已广为人知。然而,这种相互作用的确切性质以及脂蛋白随之发生的结构和/或构象改变仍有待阐明。业已证明,这种相互作用之后,LDL颗粒无法被特定细胞受体识别,并进入在不同细胞类型中运作的清道夫途径。使用主动脉PGs已证实了这些效应,但PGs也存在于血浆中,可能持续与LDL相互作用,参与脂质代谢的调节。为了评估血浆PGs诱导LDL修饰的能力,我们通过荧光光谱研究LDL组织参数的变化来探究它们之间的相互作用。通过DEAE Sephacel离子交换色谱法和Sephacryl S300凝胶过滤法,从两个不同家族中分离出血浆PGs:一种低电荷PG和一种高电荷PG。通过在1.025 - 1.045 g/ml之间进行超速离心浮选,从血脂正常供体的血浆中制备人LDL。在生理比例下将LDL与血浆PGs孵育后,通过分析二苯基己三烯(DPH)的旋转扩散速率获得稳态各向异性测量值。在我们的实验条件下,LDL与血浆低电荷PG孵育未改变DPH荧光各向异性,而用高电荷PG处理LDL则导致该参数显著降低,表明对LDL微粘度有显著影响。数据表明,PGs的电荷和糖胺聚糖组成似乎都是LDL - PG相互作用的关键因素。(摘要截短于250字)

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