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Activation of p38 Mitogen-Activated Protein Kinase in Gaucher's Disease.

Kitatani K, Wada M, Perry D, Usui T, Sun Y, Obeid LM, Yaegashi N, Grabowski GA, Hannun YA - PLoS ONE (2015)

Bottom Line: GBA1 cleaves glucosylceramide to form ceramide, an established bioactive lipid, and defects in GBA1 lead to aberrant accumulation in glucosylceramide and insufficient formation of ceramide.Most interestingly, neuronopathic Gaucher's disease type mice, but not non-neuronopathic ones, displayed significant activation of p38 and up-regulation of p38-inducible proinflammatory cytokines in brain tissues.These results raise the potential that proinflammatory responses such as p38 activation and IL-6 formation are augmented in Gaucher's disease.

View Article: PubMed Central - PubMed

Affiliation: Tohoku Medical Megabank Organization, Tohoku University, Sendai, Japan; Department of Obstetrics and Gynecology, Tohoku University Graduate School of Medicine, Sendai, Japan.

ABSTRACT
Gaucher's disease is caused by defects in acid β-glucosidase 1 (GBA1) and has been also proposed as an inflammatory disease. GBA1 cleaves glucosylceramide to form ceramide, an established bioactive lipid, and defects in GBA1 lead to aberrant accumulation in glucosylceramide and insufficient formation of ceramide. We investigated if the pro-inflammatory kinase p38 is activated in Gaucher's disease, since ceramide has been proposed to suppress p38 activation. Three Gaucher's disease mouse models were employed, and p38 was found to be activated in lung and liver tissues of all Gaucher's disease mice. Most interestingly, neuronopathic Gaucher's disease type mice, but not non-neuronopathic ones, displayed significant activation of p38 and up-regulation of p38-inducible proinflammatory cytokines in brain tissues. In addition, all type of Gaucher's disease mice also showed increases in serum IL-6. As cellular signalling is believed to represent an in vivo inflammatory phenotype in Gaucher's disease, activation of p38 and possibly its-associated formation of proinflammatory cytokines were assessed in fibroblasts established from neuronopathic Gaucher's disease mice. In mouse Gaucher's disease cells, p38 activation and IL-6 formation by TNF-α treatment were enhanced as compared to those of wild type. Furthermore, human fibroblasts from Gaucher's disease patients also displayed increases in p38 activation and IL-6 formation as comparison to healthy counterpart. These results raise the potential that proinflammatory responses such as p38 activation and IL-6 formation are augmented in Gaucher's disease.

No MeSH data available.


Related in: MedlinePlus

p38 activation and IL-6 formation in fibroblasts from Gaucher’s disease patients harboring GBA1L444P/L444P.A. Human fibroblasts were stimulated with 100 nM PMA or 25 ng/ml TNF-α for 30 min. Proteins were subjected to immunoblot analysis with antibodies specific for phospho/active-p38 (p-p38) and β-actin. Equal amounts of protein were loaded in each lane, and the representative results are shown. B. After 6 h stimulation, levels of IL-6 in culture supernatants were measured using ELISA system. IL-6 values in control, PMA and TNF-α are expressed as percentage relative to those of healthy subject, respectively. Data represent mean ± S.E. (n = 3). *, p < 0.001; **, p < 0.02; ***, p < 0.002.
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pone.0136633.g003: p38 activation and IL-6 formation in fibroblasts from Gaucher’s disease patients harboring GBA1L444P/L444P.A. Human fibroblasts were stimulated with 100 nM PMA or 25 ng/ml TNF-α for 30 min. Proteins were subjected to immunoblot analysis with antibodies specific for phospho/active-p38 (p-p38) and β-actin. Equal amounts of protein were loaded in each lane, and the representative results are shown. B. After 6 h stimulation, levels of IL-6 in culture supernatants were measured using ELISA system. IL-6 values in control, PMA and TNF-α are expressed as percentage relative to those of healthy subject, respectively. Data represent mean ± S.E. (n = 3). *, p < 0.001; **, p < 0.02; ***, p < 0.002.

Mentions: Genetic approaches and pharmacological approaches demonstrated that GBA1 defects facilitate p38 activation in vitro [16] and in vivo (above); however, whether p38 is highly activated in human Gaucher’s disease is not known. Human fibroblasts established from Gaucher’s disease patients harboring homozygous L444P were employed to assess effects of Gaucher’s disease on p38 activation and IL-6 formation. Those cells were stimulated with PMA or TNF-α, and then p38 activation was determined by immunoblotting. p38 was activated in response to TNF-α treatment in fibroblasts from healthy, whereas fibroblasts from both two Gaucher’s patients displayed significant increases in p38 activation in response to TNF-α as compared to that of healthy (Fig 3A). Similar to mouse fibroblasts, human Gaucher’s fibroblasts also displayed increased formation of IL-6 in response to individual treatments with PMA and TNF-α (Fig 3B).


Activation of p38 Mitogen-Activated Protein Kinase in Gaucher's Disease.

Kitatani K, Wada M, Perry D, Usui T, Sun Y, Obeid LM, Yaegashi N, Grabowski GA, Hannun YA - PLoS ONE (2015)

p38 activation and IL-6 formation in fibroblasts from Gaucher’s disease patients harboring GBA1L444P/L444P.A. Human fibroblasts were stimulated with 100 nM PMA or 25 ng/ml TNF-α for 30 min. Proteins were subjected to immunoblot analysis with antibodies specific for phospho/active-p38 (p-p38) and β-actin. Equal amounts of protein were loaded in each lane, and the representative results are shown. B. After 6 h stimulation, levels of IL-6 in culture supernatants were measured using ELISA system. IL-6 values in control, PMA and TNF-α are expressed as percentage relative to those of healthy subject, respectively. Data represent mean ± S.E. (n = 3). *, p < 0.001; **, p < 0.02; ***, p < 0.002.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4552301&req=5

pone.0136633.g003: p38 activation and IL-6 formation in fibroblasts from Gaucher’s disease patients harboring GBA1L444P/L444P.A. Human fibroblasts were stimulated with 100 nM PMA or 25 ng/ml TNF-α for 30 min. Proteins were subjected to immunoblot analysis with antibodies specific for phospho/active-p38 (p-p38) and β-actin. Equal amounts of protein were loaded in each lane, and the representative results are shown. B. After 6 h stimulation, levels of IL-6 in culture supernatants were measured using ELISA system. IL-6 values in control, PMA and TNF-α are expressed as percentage relative to those of healthy subject, respectively. Data represent mean ± S.E. (n = 3). *, p < 0.001; **, p < 0.02; ***, p < 0.002.
Mentions: Genetic approaches and pharmacological approaches demonstrated that GBA1 defects facilitate p38 activation in vitro [16] and in vivo (above); however, whether p38 is highly activated in human Gaucher’s disease is not known. Human fibroblasts established from Gaucher’s disease patients harboring homozygous L444P were employed to assess effects of Gaucher’s disease on p38 activation and IL-6 formation. Those cells were stimulated with PMA or TNF-α, and then p38 activation was determined by immunoblotting. p38 was activated in response to TNF-α treatment in fibroblasts from healthy, whereas fibroblasts from both two Gaucher’s patients displayed significant increases in p38 activation in response to TNF-α as compared to that of healthy (Fig 3A). Similar to mouse fibroblasts, human Gaucher’s fibroblasts also displayed increased formation of IL-6 in response to individual treatments with PMA and TNF-α (Fig 3B).

Bottom Line: GBA1 cleaves glucosylceramide to form ceramide, an established bioactive lipid, and defects in GBA1 lead to aberrant accumulation in glucosylceramide and insufficient formation of ceramide.Most interestingly, neuronopathic Gaucher's disease type mice, but not non-neuronopathic ones, displayed significant activation of p38 and up-regulation of p38-inducible proinflammatory cytokines in brain tissues.These results raise the potential that proinflammatory responses such as p38 activation and IL-6 formation are augmented in Gaucher's disease.

View Article: PubMed Central - PubMed

Affiliation: Tohoku Medical Megabank Organization, Tohoku University, Sendai, Japan; Department of Obstetrics and Gynecology, Tohoku University Graduate School of Medicine, Sendai, Japan.

ABSTRACT
Gaucher's disease is caused by defects in acid β-glucosidase 1 (GBA1) and has been also proposed as an inflammatory disease. GBA1 cleaves glucosylceramide to form ceramide, an established bioactive lipid, and defects in GBA1 lead to aberrant accumulation in glucosylceramide and insufficient formation of ceramide. We investigated if the pro-inflammatory kinase p38 is activated in Gaucher's disease, since ceramide has been proposed to suppress p38 activation. Three Gaucher's disease mouse models were employed, and p38 was found to be activated in lung and liver tissues of all Gaucher's disease mice. Most interestingly, neuronopathic Gaucher's disease type mice, but not non-neuronopathic ones, displayed significant activation of p38 and up-regulation of p38-inducible proinflammatory cytokines in brain tissues. In addition, all type of Gaucher's disease mice also showed increases in serum IL-6. As cellular signalling is believed to represent an in vivo inflammatory phenotype in Gaucher's disease, activation of p38 and possibly its-associated formation of proinflammatory cytokines were assessed in fibroblasts established from neuronopathic Gaucher's disease mice. In mouse Gaucher's disease cells, p38 activation and IL-6 formation by TNF-α treatment were enhanced as compared to those of wild type. Furthermore, human fibroblasts from Gaucher's disease patients also displayed increases in p38 activation and IL-6 formation as comparison to healthy counterpart. These results raise the potential that proinflammatory responses such as p38 activation and IL-6 formation are augmented in Gaucher's disease.

No MeSH data available.


Related in: MedlinePlus