Effect of Subchronic Intravenous Morphine Infusion and Naloxone-Precipitated Morphine Withdrawal on P-gp and Bcrp at the Rat Blood–Brain Barrier

Chaves, Catarina; Gómez-Zepeda, David; Auvity, Sylvain; Menet, Marie-Claude; Crété, Dominique; Labat, Laurence; Remião, Fernando; Cisternino, Salvatore; Declèves, Xavier

doi:10.1002/jps.24697

Cited by 22 publications

(32 citation statements)

References 44 publications

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“…; Chaves et al . ), maintaining samples at 4°C during the whole procedure. Rats were killed by CO 2 inhalation and decapitated.…”

Section: Methodsmentioning

confidence: 99%

“…Isolation of rat brain cortical microvessels Microvessels were isolated from rat brain cortex as previously described (Yousif et al 2007;Chaves et al 2016), maintaining samples at 4°C during the whole procedure. Rats were killed by CO 2 inhalation and decapitated.…”

Section: Biological Samplesmentioning

confidence: 99%

“…Protein samples were digested as previously reported (Chaves et al 2016). Briefly, denatured and alkylated proteins were precipitated using a methanol-chloroform-water system, resuspended using urea and Protease-Max detergent in Tris-HCl buffer (pH 8.5) and digested in tandem using Lys-C and Trypsin endoproteases.…”

Section: Protein Digestionmentioning

confidence: 99%

“…Therefore, molecular characterization of the endothelial cells (ECs) is important to better understand the involvement of the BBB in neuropharmacology under physiopathological situations. Targeted liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) is thus used for the absolute quantification of membrane proteins, such as ATP-binding cassette (ABC) and solute carrier transporters (Kamiie et al 2008;Hoshi et al 2013;Chaves et al 2016).…”

Section: Introductionmentioning

confidence: 99%

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Targeted unlabeled multiple reaction monitoring analysis of cell markers for the study of sample heterogeneity in isolated rat brain cortical microvessels

Gómez-Zepeda

Chaves

Taghi

et al. 2017

Journal of Neurochemistry

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Liquid chromatography coupled to tandem mass spectrometry-based targeted absolute protein quantification (in fmol of the analyte protein per μg of total protein) is employed for the molecular characterization of the blood-brain barrier using isolated brain microvessels. Nevertheless, the heterogeneity of the sample regarding the levels of different cells co-isolated within the microvessels and bovine serum albumin (BSA) contamination (from buffers) are not always evaluated. We developed an unlabeled targeted liquid chromatography coupled to tandem mass spectrometry method to survey the levels of endothelial cells (ECs), astrocytes, and pericytes, as well as BSA contaminant in rat cortical microvessels. Peptide peak identities were evaluated using a spectral library and chromatographic parameters. Sprague-Dawley rat microvessels obtained on three different days were analyzed with this method complemented by an absolute quantification multiple reaction monitoring method for transporter proteins P-gp, Bcrp, and Na /K ATPase pump using stable isotope labeled peptides as internal standard. Inter-day differences in the cell markers and BSA contamination were observed. Levels of cell markers correlated positively between each other. Then, the correlation between cell marker proteins and transporter proteins was evaluated to choose the best EC marker protein for protein quantification normalization. The membrane protein Pecam-1 showed a very high correlation with the EC-specific transporter P-gp (Pearson product-moment correlation coefficient (r) > 0.89) and moderate to high with Bcrp (r ≥ 0.77), that can be found also in pericytes and astrocytes. Therefore, Pecam-1 was selected as a marker for the normalization of the quantification of the proteins of endothelial cells.

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“…; Chaves et al . ), maintaining samples at 4°C during the whole procedure. Rats were killed by CO 2 inhalation and decapitated.…”

Section: Methodsmentioning

confidence: 99%

Section: Biological Samplesmentioning

confidence: 99%

Section: Protein Digestionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Targeted unlabeled multiple reaction monitoring analysis of cell markers for the study of sample heterogeneity in isolated rat brain cortical microvessels

Gómez-Zepeda

Chaves

Taghi

et al. 2017

Journal of Neurochemistry

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“…Diverse strategies have been devised for the ex vivo study of the brain barriers separately. Isolated microvessels can be employed for the molecular characterization of the BBB and NVU [21,22,95,168,222], used immediately for functionality studies [223][224][225][226][227], or cultured with specific media for a limited time [149]. Whole brains or specific sections (e.g., cortex) are homogenized either mechanically, employing a Potter-Evenheilm homogenizer [115,161,228] by enzymatic dissociation with collagenases that partly degrade the basement membrane [146] or using both [229]; then, the microvessels (diameter <10 µm) are separated from other cells using a density gradient and isolated from bigger vessels by sequential filtration on nylon meshes.…”

Section: Ex Vivo Modelsmentioning

confidence: 99%

ABC Transporters at the Blood–Brain Interfaces, Their Study Models, and Drug Delivery Implications in Gliomas

et al. 2019

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Drug delivery into the brain is regulated by the blood–brain interfaces. The blood–brain barrier (BBB), the blood–cerebrospinal fluid barrier (BCSFB), and the blood–arachnoid barrier (BAB) regulate the exchange of substances between the blood and brain parenchyma. These selective barriers present a high impermeability to most substances, with the selective transport of nutrients and transporters preventing the entry and accumulation of possibly toxic molecules, comprising many therapeutic drugs. Transporters of the ATP-binding cassette (ABC) superfamily have an important role in drug delivery, because they extrude a broad molecular diversity of xenobiotics, including several anticancer drugs, preventing their entry into the brain. Gliomas are the most common primary tumors diagnosed in adults, which are often characterized by a poor prognosis, notably in the case of high-grade gliomas. Therapeutic treatments frequently fail due to the difficulty of delivering drugs through the brain barriers, adding to diverse mechanisms developed by the cancer, including the overexpression or expression de novo of ABC transporters in tumoral cells and/or in the endothelial cells forming the blood–brain tumor barrier (BBTB). Many models have been developed to study the phenotype, molecular characteristics, and function of the blood–brain interfaces as well as to evaluate drug permeability into the brain. These include in vitro, in vivo, and in silico models, which together can help us to better understand their implication in drug resistance and to develop new therapeutics or delivery strategies to improve the treatment of pathologies of the central nervous system (CNS). In this review, we present the principal characteristics of the blood–brain interfaces; then, we focus on the ABC transporters present on them and their implication in drug delivery; next, we present some of the most important models used for the study of drug transport; finally, we summarize the implication of ABC transporters in glioma and the BBTB in drug resistance and the strategies to improve the delivery of CNS anticancer drugs.

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Paynantheine more effectively reverses multidrug resistance in malignant EPG85.257RDB and MCF7MX cells than morphine and speciociliatine

Abdali,

Nadipour Borujeni,

Hosseini

et al. 2024

Cell Biology International

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The possible interactions of morphine, paynantheine and speciociliatine alkaloids with ATP‐binding cassette (ABC) transporters was investigated. The compounds were docked against ABCG2 and ABCB1 to predict the binding mode of alkaloids in active binding sites. The cytotoxicity of morphine, paynantheine and speciociliatine for EPG85.257RDB and MCF7MX cells was determined and ABCB1 and ABCG2 gene and protein expression were determined. The binding score of paynantheine to ABCB1 was higher in the docking studies. Paynantheine and speciociliatine had similar binding scores to ABCB1, but higher binding scores to ABCG2 than did morphine. Paynantheine and speciociliatine were more effective against MCF7MX and EPG85.257RDB cells and showed greater cyctotoxicity in the MTT assay. The effect of morphine and paynantheine on the ABCB1 gene and protein expression suggests these compounds can reduce resistance in cancer patients, but that speciociliatine may not be a suitable candidate because of its increased ABCB1 expression while speciociliatine decreased the expression of ABCG2 in MCF7MX cells. This indicates that speciociliatine is a better candidate for reducing drug resistance in this cell line. Structural modification, drug‐metabolizing enzymes and differences in the binding sites could cause functional differences between these compounds.

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Effect of Subchronic Intravenous Morphine Infusion and Naloxone-Precipitated Morphine Withdrawal on P-gp and Bcrp at the Rat Blood–Brain Barrier

Cited by 22 publications

References 44 publications

Targeted unlabeled multiple reaction monitoring analysis of cell markers for the study of sample heterogeneity in isolated rat brain cortical microvessels

Targeted unlabeled multiple reaction monitoring analysis of cell markers for the study of sample heterogeneity in isolated rat brain cortical microvessels

ABC Transporters at the Blood–Brain Interfaces, Their Study Models, and Drug Delivery Implications in Gliomas

Paynantheine more effectively reverses multidrug resistance in malignant EPG85.257RDB and MCF7MX cells than morphine and speciociliatine

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