Comparison of gas chromatography and high-performance liquid chromatography for the analysis of probucol in plasma

Satonin, Darlene K.; Coutant, John E.

doi:10.1016/s0378-4347(00)83670-1

Cited by 39 publications

(16 citation statements)

References 3 publications

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“…Probucol is a very hydrophobic compound, and accordingly it is almost water-insoluble. Therefore, it is almost exclusively carried by lipoprotein in plasma and distributes in the membrane when cleaved to cells (37). Because almost all ABCA1 is recovered in plasma membrane, probucol is likely to act on ABCA1 in the plasma membrane.…”

Section: Discussionmentioning

confidence: 99%

“…Determination of Probucol in the Cells-Probucol in the cell lipid extracts was measured by reverse-phase high performance liquid chromatography according to the method described by Satonin and Coutant (37). The cell extracts from the medium of a 60-mm dish were dissolved in acetonitrile, hexane, 0.1 M ammonium acetate (90:6.5:3.5, v/v/v) and injected into a Deltapak C 18 reversed-phase column (150 ϫ 3.9 mm, 300 A; Waters).…”

Section: Chemicals and Reagents-probucolmentioning

confidence: 99%

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Probucol Inactivates ABCA1 in the Plasma Membrane with Respect to Its Mediation of Apolipoprotein Binding and High Density Lipoprotein Assembly and to Its Proteolytic Degradation

Chengai

Tsujita

Hayashi

et al. 2004

Journal of Biological Chemistry

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Probucol has been shown to inhibit the release of cellular lipid by helical apolipoprotein and thereby to reduce plasma high density lipoprotein. We attempted to explore the underlying mechanism for this effect in human fibroblast WI-38. Probucol inhibited the apoA-Imediated cellular lipid release and binding of apoA-I to the cells in a dose-dependent manner. It did not influence cellular uptake of low density lipoprotein, transport of cholesterol to the cell surface whether de novo synthesized or delivered as low density lipoprotein, and overall cellular content of cholesterol, although biosynthesis of lipids from acetate was somewhat increased. Probucol did not affect the mRNA level of ABCA1, and ABCA1 was recovered along with marker proteins for plasma membrane regardless of the presence of probucol. However, the protein level of ABCA1 increased, and the rate of its decay in the presence of cycloheximide was slower in the probucol-treated cells. ABCA1 in the probucol-treated cells was resistant to digestion by calpain but not by trypsin. We concluded that probucol inactivates ABCA1 in the plasma membrane with respect to its function in mediating binding of and lipid release by apolipoprotein and with respect to proteolytic degradation by calpain.Cholesterol is an essential molecule for animal cells to maintain and regulate function and structure of the biomembrane. It is synthesized in most somatic cells, whereas its catabolic site is limited to the liver and to the steroidogenic cells except for partial hydroxylation in some somatic cells. Accordingly, cholesterol is removed from the cells and transported to the liver for its conversion to bile acids, and this is one of the essential events in cholesterol homeostasis for the body and for the cells (1). High density lipoprotein (HDL) 1 is believed to play a central role in this system, and this is thought to be one of the antiatherogenic characteristics of HDL. This reaction takes place through at least two distinct mechanisms: 1) physicochemical release of cholesterol from the cell surface, which is driven by cholesterol esterification on HDL, and 2) the apolipoprotein-mediated pathway to remove cellular cholesterol and phospholipid to generate new HDL particles (2). HDL thus plays a central role in both mechanisms.Apolipoprotein-dependent cellular cholesterol release is absent in fibroblasts from patients with Tangier disease (3, 4), and mutations in the gene encoding the ATP-binding cassette transporter A1 (ABCA1) are the underlying cause of this disease (5-9). On the other hand, in vitro overexpression of functional ABCA1 in the cells (10, 11) and induction of ABCA1 expression by cyclic AMP analogues (12, 13) or by the ligands for the liver X receptor or retinoid X receptor (14, 15) enhanced the release of cellular cholesterol and phospholipid by apolipoprotein. The transgenic mice for ABCA1 had a significant increase in plasma HDL (16,17). These results indicate that this protein is a regulating factor for the plasma HDL level through generation of HDL b...

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Section: Discussionmentioning

confidence: 99%

Section: Chemicals and Reagents-probucolmentioning

confidence: 99%

Probucol Inactivates ABCA1 in the Plasma Membrane with Respect to Its Mediation of Apolipoprotein Binding and High Density Lipoprotein Assembly and to Its Proteolytic Degradation

Chengai

Tsujita

Hayashi

et al. 2004

Journal of Biological Chemistry

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“…The sample was also used for probucol analysis using an HPLC system (22). Protein was measured with the bicinchoninic acid method using BSA as a standard.…”

Section: Other Methodsmentioning

confidence: 99%

On the hepatic mechanism of HDL assembly by the ABCA1/apoA-I pathway

Tsujita

Abe-Dohmae

et al. 2005

Journal of Lipid Research

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The mechanism for the assembly of HDL with cellular lipid by ABCA1 and helical apolipoprotein was investigated in hepatocytes. Both HepG2 cells and mouse primary culture hepatocytes produced HDL with apolipoprotein A-I (apoA-I) whether endogenously synthesized or exogenously provided. Probucol, an ABCA1 inactivator, inhibited these reactions, as well as the reversible binding of apoA-I to HepG2. Primary cultured hepatocytes of ABCA1-deficient mice also lacked HDL production regardless of the presence of exogenous apoA-I. HepG2 cells secreted apoA-I into the medium even when ABCA1 was inactivated by probucol, but it was all in a free form as HDL production was inhibited. When a lipid-free apoA-I-specific monoclonal antibody, 725-1E2, was present in the culture medium, production of HDL was suppressed, whether with endogenous or exogenously added apoA-I, and the antibody did not influence HDL already produced by HepG2 cells. We conclude that the main mechanism for HDL assembly by endogenous apoA-I in HepG2 cells is an autocrine-like reaction in which apoA-I is secreted and then interacts with cellular ABCA1 to generate HDL. -Tsujita, M., C-A. Wu, S. AbeDohmae, S. Usui, M. Okazaki, and S. Yokoyama. On the hepatic mechanism of HDL assembly by the ABCA1/apoA-I pathway. J. Lipid Res. 2005. 46: 154-162.

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“…1718 The concentration of probucol was determined by high-performance liquid chromatography at Wisconsin Analytical Research Services, Madison, Wis. 19 Total protein content of the LDL preparation was determined using bovine serum albumin as the standard. 20 …”

Section: Other Analysesmentioning

confidence: 99%

Influence of probucol on enhanced LDL oxidation after fish oil treatment of hypertriglyceridemic patients.

Lussier‐Cacan

Dubreuil-Quidoz

Roederer

et al. 1993

Arterioscler Thromb

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The susceptibility of low-density lipoprotein (LDL) to oxidation was studied in hypertriglyceridemic men (5 with type III and 5 with type IV) at baseline on a low-saturated-fat, low-cholesterol diet, after 6 weeks of dietary supplementation with fish oil (Promega, 12 g/d), and after 6 weeks of fish oil combined with probucol (500 mg BID). The relative content of n-3 polyunsaturated fatty acids in plasma and LDL was increased during the two treatment periods, and a low a-tocopherol to n-3 polyunsaturated fatty acids ratio was observed. Plasma thiobarbituric acid-reactive substances (TBARS) levels were unchanged after 6 weeks of fish oil, but the ratio of lipid peroxides to the reduced triglyceride (TG) levels (MDA: TG) was significantly higher (P<.01). Addition of probucol lowered both absolute levels of TBARS (P<.01) and the MDA to TG ratio (P<.001). The susceptibility of LDL to Cu 2+ -catalyzed oxidation was evaluated over a 5-hour time course by determining TBARS formation, free amino group levels, and changes in LDL electrophoretic mobility. TBARS levels that were higher in native LDL (1.019«/< 1.050 g/mL) after 6 weeks offish oil than at baseline (P<.0l) were reduced 52.3±11.3% by the addition of probucol (P<.001). With fish oil alone, TBARS production after exposure of LDL to Cu

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Comparison of gas chromatography and high-performance liquid chromatography for the analysis of probucol in plasma

Cited by 39 publications

References 3 publications

Probucol Inactivates ABCA1 in the Plasma Membrane with Respect to Its Mediation of Apolipoprotein Binding and High Density Lipoprotein Assembly and to Its Proteolytic Degradation

Probucol Inactivates ABCA1 in the Plasma Membrane with Respect to Its Mediation of Apolipoprotein Binding and High Density Lipoprotein Assembly and to Its Proteolytic Degradation

On the hepatic mechanism of HDL assembly by the ABCA1/apoA-I pathway

Influence of probucol on enhanced LDL oxidation after fish oil treatment of hypertriglyceridemic patients.

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