Limits...
Aquaporin-3 potentiates allergic airway inflammation in ovalbumin-induced murine asthma.

Ikezoe K, Oga T, Honda T, Hara-Chikuma M, Ma X, Tsuruyama T, Uno K, Fuchikami J, Tanizawa K, Handa T, Taguchi Y, Verkman AS, Narumiya S, Mishima M, Chin K - Sci Rep (2016)

Bottom Line: Aquaporin-3 (AQP3) is a small transmembrane water/glycerol channel that may facilitate the membrane uptake of hydrogen peroxide (H2O2).Additionally, in vivo and vitro experiments indicated that AQP3 induced the production of some chemokines such as CCL24 and CCL22 through regulating the amount of cellular H2O2 in M2 polarized alveolar macrophages.These results imply a critical role of AQP3 in asthma, and AQP3 may be a novel therapeutic target.

View Article: PubMed Central - PubMed

Affiliation: Department of Respiratory Medicine, Kyoto University Graduate School of Medicine, Sakyo-ku, Kyoto 606-8507, Japan.

ABSTRACT
Oxidative stress plays a pivotal role in the pathogenesis of asthma. Aquaporin-3 (AQP3) is a small transmembrane water/glycerol channel that may facilitate the membrane uptake of hydrogen peroxide (H2O2). Here we report that AQP3 potentiates ovalbumin (OVA)-induced murine asthma by mediating both chemokine production from alveolar macrophages and T cell trafficking. AQP3 deficient (AQP3(-/-)) mice exhibited significantly reduced airway inflammation compared to wild-type mice. Adoptive transfer experiments showed reduced airway eosinophilic inflammation in mice receiving OVA-sensitized splenocytes from AQP3(-/-) mice compared with wild-type mice after OVA challenge, consistently with fewer CD4(+) T cells from AQP3(-/-) mice migrating to the lung than from wild-type mice. Additionally, in vivo and vitro experiments indicated that AQP3 induced the production of some chemokines such as CCL24 and CCL22 through regulating the amount of cellular H2O2 in M2 polarized alveolar macrophages. These results imply a critical role of AQP3 in asthma, and AQP3 may be a novel therapeutic target.

No MeSH data available.


Related in: MedlinePlus

AQP3 regulates chemokine production from M2 polarized AMs through affecting the amount of cellular H2O2.(a) H2O2 permeability in naive AMs. Sorted AMs from naïve WT and AQP3−/− mice were incubated with/without H2O2 (100 μM) for 15 s. Cellular H2O2 levels were determined using CM-H2DCFDA reagent by flow cytometry analysis. Mean fluorescence intensity (MFI) of CM-H2DCFDA is shown (n = 3 for each group). (b) Intracellular H2O2 levels in IL-4/13-stimulated AMs. Sorted AMs from naïve WT and AQP3−/− mice were incubated with IL-4/13 for 24 h, and cellular H2O2 levels were determined (n = 4 for each group). As the numbers of AMs were small, we combined the results of 2 independent experiments. Ratios of MFI in each sample to MFI in naïve non-stimulated WT AMs are shown. (c) Cellular H2O2 levels in IL-4/13 stimulated AMs with/without PEG-catalase. Sorted AMs from naïve WT mice were incubated with 250 U/ml PEG-catalase for 1 h and followed with IL-4/13 for 24 h. Cellular H2O2 levels in AMs determined using CM-H2DCFDA reagent (n = 4–5 for each group). (d,e) mRNA expression levels of Ccl24 and Ccl22 (d), and Arg1 and Retnla (e) in IL-4/13 stimulated AMs with/without PEG-catalase were measured by real-time RT-PCR (n = 4 for each group). **P < 0.01, *P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4863152&req=5

f7: AQP3 regulates chemokine production from M2 polarized AMs through affecting the amount of cellular H2O2.(a) H2O2 permeability in naive AMs. Sorted AMs from naïve WT and AQP3−/− mice were incubated with/without H2O2 (100 μM) for 15 s. Cellular H2O2 levels were determined using CM-H2DCFDA reagent by flow cytometry analysis. Mean fluorescence intensity (MFI) of CM-H2DCFDA is shown (n = 3 for each group). (b) Intracellular H2O2 levels in IL-4/13-stimulated AMs. Sorted AMs from naïve WT and AQP3−/− mice were incubated with IL-4/13 for 24 h, and cellular H2O2 levels were determined (n = 4 for each group). As the numbers of AMs were small, we combined the results of 2 independent experiments. Ratios of MFI in each sample to MFI in naïve non-stimulated WT AMs are shown. (c) Cellular H2O2 levels in IL-4/13 stimulated AMs with/without PEG-catalase. Sorted AMs from naïve WT mice were incubated with 250 U/ml PEG-catalase for 1 h and followed with IL-4/13 for 24 h. Cellular H2O2 levels in AMs determined using CM-H2DCFDA reagent (n = 4–5 for each group). (d,e) mRNA expression levels of Ccl24 and Ccl22 (d), and Arg1 and Retnla (e) in IL-4/13 stimulated AMs with/without PEG-catalase were measured by real-time RT-PCR (n = 4 for each group). **P < 0.01, *P < 0.05.

Mentions: We anticipated that AQP3 would affect chemokine production from AMs by modulating the amount of cellular H2O2. Firstly, we measured intracellular H2O2 levels in AMs using 5-(and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate acetyl ester (CM-H2DCFDA) fluorescence. We found that cellular H2O2 levels were lower in naïve AQP3−/− AMs than in naïve WT AMs without and with exogenous H2O2 supplementation for 15 s (Fig. 7a). Stimulation of AMs with IL-4/13 for 24 h increased cellular H2O2 levels in both AQP3−/− and WT AMs, but its enhancement was more reduced in AQP3−/− AMs (Fig. 7b). Thus, AQP3 was considered to be involved in regulating cellular H2O2 concentration in AMs under Th2 environment.


Aquaporin-3 potentiates allergic airway inflammation in ovalbumin-induced murine asthma.

Ikezoe K, Oga T, Honda T, Hara-Chikuma M, Ma X, Tsuruyama T, Uno K, Fuchikami J, Tanizawa K, Handa T, Taguchi Y, Verkman AS, Narumiya S, Mishima M, Chin K - Sci Rep (2016)

AQP3 regulates chemokine production from M2 polarized AMs through affecting the amount of cellular H2O2.(a) H2O2 permeability in naive AMs. Sorted AMs from naïve WT and AQP3−/− mice were incubated with/without H2O2 (100 μM) for 15 s. Cellular H2O2 levels were determined using CM-H2DCFDA reagent by flow cytometry analysis. Mean fluorescence intensity (MFI) of CM-H2DCFDA is shown (n = 3 for each group). (b) Intracellular H2O2 levels in IL-4/13-stimulated AMs. Sorted AMs from naïve WT and AQP3−/− mice were incubated with IL-4/13 for 24 h, and cellular H2O2 levels were determined (n = 4 for each group). As the numbers of AMs were small, we combined the results of 2 independent experiments. Ratios of MFI in each sample to MFI in naïve non-stimulated WT AMs are shown. (c) Cellular H2O2 levels in IL-4/13 stimulated AMs with/without PEG-catalase. Sorted AMs from naïve WT mice were incubated with 250 U/ml PEG-catalase for 1 h and followed with IL-4/13 for 24 h. Cellular H2O2 levels in AMs determined using CM-H2DCFDA reagent (n = 4–5 for each group). (d,e) mRNA expression levels of Ccl24 and Ccl22 (d), and Arg1 and Retnla (e) in IL-4/13 stimulated AMs with/without PEG-catalase were measured by real-time RT-PCR (n = 4 for each group). **P < 0.01, *P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4863152&req=5

f7: AQP3 regulates chemokine production from M2 polarized AMs through affecting the amount of cellular H2O2.(a) H2O2 permeability in naive AMs. Sorted AMs from naïve WT and AQP3−/− mice were incubated with/without H2O2 (100 μM) for 15 s. Cellular H2O2 levels were determined using CM-H2DCFDA reagent by flow cytometry analysis. Mean fluorescence intensity (MFI) of CM-H2DCFDA is shown (n = 3 for each group). (b) Intracellular H2O2 levels in IL-4/13-stimulated AMs. Sorted AMs from naïve WT and AQP3−/− mice were incubated with IL-4/13 for 24 h, and cellular H2O2 levels were determined (n = 4 for each group). As the numbers of AMs were small, we combined the results of 2 independent experiments. Ratios of MFI in each sample to MFI in naïve non-stimulated WT AMs are shown. (c) Cellular H2O2 levels in IL-4/13 stimulated AMs with/without PEG-catalase. Sorted AMs from naïve WT mice were incubated with 250 U/ml PEG-catalase for 1 h and followed with IL-4/13 for 24 h. Cellular H2O2 levels in AMs determined using CM-H2DCFDA reagent (n = 4–5 for each group). (d,e) mRNA expression levels of Ccl24 and Ccl22 (d), and Arg1 and Retnla (e) in IL-4/13 stimulated AMs with/without PEG-catalase were measured by real-time RT-PCR (n = 4 for each group). **P < 0.01, *P < 0.05.
Mentions: We anticipated that AQP3 would affect chemokine production from AMs by modulating the amount of cellular H2O2. Firstly, we measured intracellular H2O2 levels in AMs using 5-(and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate acetyl ester (CM-H2DCFDA) fluorescence. We found that cellular H2O2 levels were lower in naïve AQP3−/− AMs than in naïve WT AMs without and with exogenous H2O2 supplementation for 15 s (Fig. 7a). Stimulation of AMs with IL-4/13 for 24 h increased cellular H2O2 levels in both AQP3−/− and WT AMs, but its enhancement was more reduced in AQP3−/− AMs (Fig. 7b). Thus, AQP3 was considered to be involved in regulating cellular H2O2 concentration in AMs under Th2 environment.

Bottom Line: Aquaporin-3 (AQP3) is a small transmembrane water/glycerol channel that may facilitate the membrane uptake of hydrogen peroxide (H2O2).Additionally, in vivo and vitro experiments indicated that AQP3 induced the production of some chemokines such as CCL24 and CCL22 through regulating the amount of cellular H2O2 in M2 polarized alveolar macrophages.These results imply a critical role of AQP3 in asthma, and AQP3 may be a novel therapeutic target.

View Article: PubMed Central - PubMed

Affiliation: Department of Respiratory Medicine, Kyoto University Graduate School of Medicine, Sakyo-ku, Kyoto 606-8507, Japan.

ABSTRACT
Oxidative stress plays a pivotal role in the pathogenesis of asthma. Aquaporin-3 (AQP3) is a small transmembrane water/glycerol channel that may facilitate the membrane uptake of hydrogen peroxide (H2O2). Here we report that AQP3 potentiates ovalbumin (OVA)-induced murine asthma by mediating both chemokine production from alveolar macrophages and T cell trafficking. AQP3 deficient (AQP3(-/-)) mice exhibited significantly reduced airway inflammation compared to wild-type mice. Adoptive transfer experiments showed reduced airway eosinophilic inflammation in mice receiving OVA-sensitized splenocytes from AQP3(-/-) mice compared with wild-type mice after OVA challenge, consistently with fewer CD4(+) T cells from AQP3(-/-) mice migrating to the lung than from wild-type mice. Additionally, in vivo and vitro experiments indicated that AQP3 induced the production of some chemokines such as CCL24 and CCL22 through regulating the amount of cellular H2O2 in M2 polarized alveolar macrophages. These results imply a critical role of AQP3 in asthma, and AQP3 may be a novel therapeutic target.

No MeSH data available.


Related in: MedlinePlus