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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

T-lymphocytes with high P-gp expression dominated the PBMC of TNBS-induced mice.Isolated PBMC were incubated with FITC-conjugated anti-mouse P-gp antibody together with APC-conjugated anti-mouse CD3, CD14 or CD19 antibodies. The data were acquired on a BD FACSVerse flow cytometer and analyzed. (A) The cells were first gated crudely for monocytes (Gate1) or lymphocytes (Gate2) using SSC-H vs. FSC-H. (B) The cells in Gate1 or Gate2 were further quadrant-gated by comparing them with the negative-stained group, and FITC/APC double-positive staining (upper right in quadrant gating) was considered to represent positive P-gp expression in T cells (CD3+), B cells (CD19+) and monocytes (CD14+). (C) By multiplying the percentage of APC-positive (upper left and upper right quadrant gating) cells in Gate1/Gate2 by the percentage of Gate1/Gate2 in PBMC, the relative contents of T-lymphocytes, B-lymphocytes and monocytes in the PBMC of colitis mice were determined. (D) By multiplying the percentage of FITC/APC double-positive (upper right in quadrant gating) cells in Gate1/Gate2 by the percentage of Gate1/Gate2 in PBMC, the percentages of positive P-gp expressing T-lymphocytes, B-lymphocytes and monocytes in the PBMC of colitis mice were determined.
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f3: T-lymphocytes with high P-gp expression dominated the PBMC of TNBS-induced mice.Isolated PBMC were incubated with FITC-conjugated anti-mouse P-gp antibody together with APC-conjugated anti-mouse CD3, CD14 or CD19 antibodies. The data were acquired on a BD FACSVerse flow cytometer and analyzed. (A) The cells were first gated crudely for monocytes (Gate1) or lymphocytes (Gate2) using SSC-H vs. FSC-H. (B) The cells in Gate1 or Gate2 were further quadrant-gated by comparing them with the negative-stained group, and FITC/APC double-positive staining (upper right in quadrant gating) was considered to represent positive P-gp expression in T cells (CD3+), B cells (CD19+) and monocytes (CD14+). (C) By multiplying the percentage of APC-positive (upper left and upper right quadrant gating) cells in Gate1/Gate2 by the percentage of Gate1/Gate2 in PBMC, the relative contents of T-lymphocytes, B-lymphocytes and monocytes in the PBMC of colitis mice were determined. (D) By multiplying the percentage of FITC/APC double-positive (upper right in quadrant gating) cells in Gate1/Gate2 by the percentage of Gate1/Gate2 in PBMC, the percentages of positive P-gp expressing T-lymphocytes, B-lymphocytes and monocytes in the PBMC of colitis mice were determined.

Mentions: As seen in Fig 3A, PBMC were gated crudely for the monocyte group (Gate 1, 20.08%) and the lymphocyte group (Gate 2, 73.88%) by using SSC-H vs. FSC-H. Then, the cells in each gate were further identified by APC/FITC double-staining to certify P-gp positive expression in a subset of PBMC. Focusing on the upper right quadrant gating in Fig 3B, the percentages of positive P-gp-expressing T-lymphocytes (CD3+) and monocytes (CD14+) were increased significantly from 41.0% to 82.6% and from 6.8% to 19.6%, respectively, when colitis was induced in normal mice. However, the percentage of P-gp positive expression in B-lymphocytes (CD19+) was not changed markedly.


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)

T-lymphocytes with high P-gp expression dominated the PBMC of TNBS-induced mice.Isolated PBMC were incubated with FITC-conjugated anti-mouse P-gp antibody together with APC-conjugated anti-mouse CD3, CD14 or CD19 antibodies. The data were acquired on a BD FACSVerse flow cytometer and analyzed. (A) The cells were first gated crudely for monocytes (Gate1) or lymphocytes (Gate2) using SSC-H vs. FSC-H. (B) The cells in Gate1 or Gate2 were further quadrant-gated by comparing them with the negative-stained group, and FITC/APC double-positive staining (upper right in quadrant gating) was considered to represent positive P-gp expression in T cells (CD3+), B cells (CD19+) and monocytes (CD14+). (C) By multiplying the percentage of APC-positive (upper left and upper right quadrant gating) cells in Gate1/Gate2 by the percentage of Gate1/Gate2 in PBMC, the relative contents of T-lymphocytes, B-lymphocytes and monocytes in the PBMC of colitis mice were determined. (D) By multiplying the percentage of FITC/APC double-positive (upper right in quadrant gating) cells in Gate1/Gate2 by the percentage of Gate1/Gate2 in PBMC, the percentages of positive P-gp expressing T-lymphocytes, B-lymphocytes and monocytes in the PBMC of colitis mice were determined.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4555107&req=5

f3: T-lymphocytes with high P-gp expression dominated the PBMC of TNBS-induced mice.Isolated PBMC were incubated with FITC-conjugated anti-mouse P-gp antibody together with APC-conjugated anti-mouse CD3, CD14 or CD19 antibodies. The data were acquired on a BD FACSVerse flow cytometer and analyzed. (A) The cells were first gated crudely for monocytes (Gate1) or lymphocytes (Gate2) using SSC-H vs. FSC-H. (B) The cells in Gate1 or Gate2 were further quadrant-gated by comparing them with the negative-stained group, and FITC/APC double-positive staining (upper right in quadrant gating) was considered to represent positive P-gp expression in T cells (CD3+), B cells (CD19+) and monocytes (CD14+). (C) By multiplying the percentage of APC-positive (upper left and upper right quadrant gating) cells in Gate1/Gate2 by the percentage of Gate1/Gate2 in PBMC, the relative contents of T-lymphocytes, B-lymphocytes and monocytes in the PBMC of colitis mice were determined. (D) By multiplying the percentage of FITC/APC double-positive (upper right in quadrant gating) cells in Gate1/Gate2 by the percentage of Gate1/Gate2 in PBMC, the percentages of positive P-gp expressing T-lymphocytes, B-lymphocytes and monocytes in the PBMC of colitis mice were determined.
Mentions: As seen in Fig 3A, PBMC were gated crudely for the monocyte group (Gate 1, 20.08%) and the lymphocyte group (Gate 2, 73.88%) by using SSC-H vs. FSC-H. Then, the cells in each gate were further identified by APC/FITC double-staining to certify P-gp positive expression in a subset of PBMC. Focusing on the upper right quadrant gating in Fig 3B, the percentages of positive P-gp-expressing T-lymphocytes (CD3+) and monocytes (CD14+) were increased significantly from 41.0% to 82.6% and from 6.8% to 19.6%, respectively, when colitis was induced in normal mice. However, the percentage of P-gp positive expression in B-lymphocytes (CD19+) was not changed markedly.

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