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Chronic inflammation up-regulates P-gp in peripheral mononuclear blood cells via the STAT3/Nf-κb pathway in 2,4,6-trinitrobenzene sulfonic acid-induced colitis mice.

Liu J, Zhou F, Chen Q, Kang A, Lu M, Liu W, Zang X, Wang G, Zhang J - Sci Rep (2015)

Bottom Line: This resistance can be divided into intrinsic resistance and acquired resistance.The results revealed reduced retention of cyclosporine A in PMBC over-expressing P-gp in a TNBS-treated group and enhanced secretion of the cytokines IL-1β, IL-6, IL-17, and TNF-α as well as LPS in plasma.These cytokines and LPS can induce P-gp expression through the STAT3/Nf-κb pathway, contributing to a decrease of cyclosporine A retention, which can be reversed by the application of a P-gp inhibitor.

View Article: PubMed Central - PubMed

Affiliation: Key Lab of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, Jiangsu, China.

ABSTRACT
Patients with inflammatory bowel diseases, including Crohn's disease and ulcerative colitis, often suffer drug intolerance. This resistance can be divided into intrinsic resistance and acquired resistance. Although there is agreement on acquired resistance, studies regarding intrinsic resistance have demonstrated inconsistencies, especially for Crohn's disease. For this reason, an animal model of Crohn's disease was induced with 2,4,6-trinitrobenzene sulfonic acid solution (TNBS), and intrinsic resistance was analyzed by measuring the function and expression of P-glycoprotein (P-gp) in peripheral mononuclear blood cells (PMBC), followed by mechanistic studies. The results revealed reduced retention of cyclosporine A in PMBC over-expressing P-gp in a TNBS-treated group and enhanced secretion of the cytokines IL-1β, IL-6, IL-17, and TNF-α as well as LPS in plasma. These cytokines and LPS can induce P-gp expression through the STAT3/Nf-κb pathway, contributing to a decrease of cyclosporine A retention, which can be reversed by the application of a P-gp inhibitor. Our results demonstrated that the sustained chronic inflammation could induce the intrinsic resistance presented as P-gp over-expression in PBMC in Crohn's disease through STAT3/Nf-κb pathway and this resistance might be reversed by combinational usage of P-gp inhibitors.

No MeSH data available.


Related in: MedlinePlus

Elevated P-gp levels in monocytes and T-lymphocytes affected the intracellular pharmacokinetics of Cys A.THP-1 monocytes were pre-incubated with LPS for 72 h for P-gp induction, which was confirmed by an intracellular retention assay of the P-gp substrate probe Rho 123 (A), and the P-gp substrate immunosuppressant Cys A (B) in the presence or absence of the P-gp inhibitor LY335975. CCRF-CEM MDR1 lymphocytes were constructed from the CCRF-CEM parental cell line by lentiviral transfection. Their P-gp protein levels were analyzed by flow cytometry using a FITC-conjugated anti-P-gp antibody (C) and were quantified based on mean fluorescence intensity (D). The mRNA levels of P-gp were also measured by qPCR using actin as an internal control (E). The function of P-gp was evaluated through an intracellular retention assay using Rho 123 as a P-gp substrate probe (F) and Cys A as a P-gp substrate immunosuppressant (G) in the absence or presence of the P-gp inhibitor LY335979. (H) The uptake kinetics of the immunosuppressant Cys A in P-gp-overexpressing lymphocytes were determined by incubating CCRF-CEM MDR1 cells with Cys A in the absence or presence of the P-gp inhibitor LY335979. All of the data are expressed as the mean ± S.E.M.; *p < 0.05, **p < 0.01 compared with the corresponding control group; n = 3.
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f8: Elevated P-gp levels in monocytes and T-lymphocytes affected the intracellular pharmacokinetics of Cys A.THP-1 monocytes were pre-incubated with LPS for 72 h for P-gp induction, which was confirmed by an intracellular retention assay of the P-gp substrate probe Rho 123 (A), and the P-gp substrate immunosuppressant Cys A (B) in the presence or absence of the P-gp inhibitor LY335975. CCRF-CEM MDR1 lymphocytes were constructed from the CCRF-CEM parental cell line by lentiviral transfection. Their P-gp protein levels were analyzed by flow cytometry using a FITC-conjugated anti-P-gp antibody (C) and were quantified based on mean fluorescence intensity (D). The mRNA levels of P-gp were also measured by qPCR using actin as an internal control (E). The function of P-gp was evaluated through an intracellular retention assay using Rho 123 as a P-gp substrate probe (F) and Cys A as a P-gp substrate immunosuppressant (G) in the absence or presence of the P-gp inhibitor LY335979. (H) The uptake kinetics of the immunosuppressant Cys A in P-gp-overexpressing lymphocytes were determined by incubating CCRF-CEM MDR1 cells with Cys A in the absence or presence of the P-gp inhibitor LY335979. All of the data are expressed as the mean ± S.E.M.; *p < 0.05, **p < 0.01 compared with the corresponding control group; n = 3.

Mentions: As seen in Fig 8A, inflammation-mediated P-gp-overexpressing monocytes were developed by incubating THP-1 cells with LPS. Rho 123 as an indicator of P-gp function; its intracellular accumulation was markedly decreased in LPS-treated THP-1 cells and could be reversed by the application of P-gp inhibitor. For the P-gp substrate drug Cys A, LPS-mediated inflammatory monocytes exhibited lower intracellular accumulation of Cys A, which was only 69% of that in the control cells and could be reversed by the application of P-gp inhibitor (Fig. 8B).


Chronic inflammation up-regulates P-gp in peripheral mononuclear blood cells via the STAT3/Nf-κb pathway in 2,4,6-trinitrobenzene sulfonic acid-induced colitis mice.

Liu J, Zhou F, Chen Q, Kang A, Lu M, Liu W, Zang X, Wang G, Zhang J - Sci Rep (2015)

Elevated P-gp levels in monocytes and T-lymphocytes affected the intracellular pharmacokinetics of Cys A.THP-1 monocytes were pre-incubated with LPS for 72 h for P-gp induction, which was confirmed by an intracellular retention assay of the P-gp substrate probe Rho 123 (A), and the P-gp substrate immunosuppressant Cys A (B) in the presence or absence of the P-gp inhibitor LY335975. CCRF-CEM MDR1 lymphocytes were constructed from the CCRF-CEM parental cell line by lentiviral transfection. Their P-gp protein levels were analyzed by flow cytometry using a FITC-conjugated anti-P-gp antibody (C) and were quantified based on mean fluorescence intensity (D). The mRNA levels of P-gp were also measured by qPCR using actin as an internal control (E). The function of P-gp was evaluated through an intracellular retention assay using Rho 123 as a P-gp substrate probe (F) and Cys A as a P-gp substrate immunosuppressant (G) in the absence or presence of the P-gp inhibitor LY335979. (H) The uptake kinetics of the immunosuppressant Cys A in P-gp-overexpressing lymphocytes were determined by incubating CCRF-CEM MDR1 cells with Cys A in the absence or presence of the P-gp inhibitor LY335979. All of the data are expressed as the mean ± S.E.M.; *p < 0.05, **p < 0.01 compared with the corresponding control group; n = 3.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4555107&req=5

f8: Elevated P-gp levels in monocytes and T-lymphocytes affected the intracellular pharmacokinetics of Cys A.THP-1 monocytes were pre-incubated with LPS for 72 h for P-gp induction, which was confirmed by an intracellular retention assay of the P-gp substrate probe Rho 123 (A), and the P-gp substrate immunosuppressant Cys A (B) in the presence or absence of the P-gp inhibitor LY335975. CCRF-CEM MDR1 lymphocytes were constructed from the CCRF-CEM parental cell line by lentiviral transfection. Their P-gp protein levels were analyzed by flow cytometry using a FITC-conjugated anti-P-gp antibody (C) and were quantified based on mean fluorescence intensity (D). The mRNA levels of P-gp were also measured by qPCR using actin as an internal control (E). The function of P-gp was evaluated through an intracellular retention assay using Rho 123 as a P-gp substrate probe (F) and Cys A as a P-gp substrate immunosuppressant (G) in the absence or presence of the P-gp inhibitor LY335979. (H) The uptake kinetics of the immunosuppressant Cys A in P-gp-overexpressing lymphocytes were determined by incubating CCRF-CEM MDR1 cells with Cys A in the absence or presence of the P-gp inhibitor LY335979. All of the data are expressed as the mean ± S.E.M.; *p < 0.05, **p < 0.01 compared with the corresponding control group; n = 3.
Mentions: As seen in Fig 8A, inflammation-mediated P-gp-overexpressing monocytes were developed by incubating THP-1 cells with LPS. Rho 123 as an indicator of P-gp function; its intracellular accumulation was markedly decreased in LPS-treated THP-1 cells and could be reversed by the application of P-gp inhibitor. For the P-gp substrate drug Cys A, LPS-mediated inflammatory monocytes exhibited lower intracellular accumulation of Cys A, which was only 69% of that in the control cells and could be reversed by the application of P-gp inhibitor (Fig. 8B).

Bottom Line: This resistance can be divided into intrinsic resistance and acquired resistance.The results revealed reduced retention of cyclosporine A in PMBC over-expressing P-gp in a TNBS-treated group and enhanced secretion of the cytokines IL-1β, IL-6, IL-17, and TNF-α as well as LPS in plasma.These cytokines and LPS can induce P-gp expression through the STAT3/Nf-κb pathway, contributing to a decrease of cyclosporine A retention, which can be reversed by the application of a P-gp inhibitor.

View Article: PubMed Central - PubMed

Affiliation: Key Lab of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, Jiangsu, China.

ABSTRACT
Patients with inflammatory bowel diseases, including Crohn's disease and ulcerative colitis, often suffer drug intolerance. This resistance can be divided into intrinsic resistance and acquired resistance. Although there is agreement on acquired resistance, studies regarding intrinsic resistance have demonstrated inconsistencies, especially for Crohn's disease. For this reason, an animal model of Crohn's disease was induced with 2,4,6-trinitrobenzene sulfonic acid solution (TNBS), and intrinsic resistance was analyzed by measuring the function and expression of P-glycoprotein (P-gp) in peripheral mononuclear blood cells (PMBC), followed by mechanistic studies. The results revealed reduced retention of cyclosporine A in PMBC over-expressing P-gp in a TNBS-treated group and enhanced secretion of the cytokines IL-1β, IL-6, IL-17, and TNF-α as well as LPS in plasma. These cytokines and LPS can induce P-gp expression through the STAT3/Nf-κb pathway, contributing to a decrease of cyclosporine A retention, which can be reversed by the application of a P-gp inhibitor. Our results demonstrated that the sustained chronic inflammation could induce the intrinsic resistance presented as P-gp over-expression in PBMC in Crohn's disease through STAT3/Nf-κb pathway and this resistance might be reversed by combinational usage of P-gp inhibitors.

No MeSH data available.


Related in: MedlinePlus