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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 a Gaucher’s disease mouse model.Proteins were extracted from tissues (brain, lung, and liver) of wild type mice or Gaucher disease mouse models (V394L, D409H, and V394L/PS-NA) and then subjected to immunoblot analysis with antibodies specific for phospho- p38 and β-actin. Equal amounts of protein were loaded in each lane, and the representative results of brain tissues are shown (A). Amounts of active/phospho-p38 were estimated by measuring the density of bands of phospho-p38 and expressed as arbitrary units (B). The data represent mean ± S.E. (n = 4–7). *, p < 0.02; **, p < 0.05. (C) mRNA was extracted from brain tissues from wild type and neuropathic Gaucher disease model V394L/PS-NA mice, and mRNAs of p38 isoforms and IL-6 were determined by the quantitative real time PCR. The data represent mean ± S.E. (n = 5). TNF-α, p < 0.0006 (*). (D) Serum IL-6 levels from wild type mice or Gaucher disease mouse models (V394L, D409H, and V394L/PS-NA) were determined by the ELISA system. The data represent mean ± S.E. Wild type, n = 21; V394L, n = 10; D409H, n = 7; V394L/PS-NA, n = 7. Wt vs V394L, p < 0.0006 (*); Wt vs D409, p < 0.03 (**); Wt vs V394L/PS-NA, p < 0.05 (***).
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pone.0136633.g001: p38 activation and IL-6 formation in a Gaucher’s disease mouse model.Proteins were extracted from tissues (brain, lung, and liver) of wild type mice or Gaucher disease mouse models (V394L, D409H, and V394L/PS-NA) and then subjected to immunoblot analysis with antibodies specific for phospho- p38 and β-actin. Equal amounts of protein were loaded in each lane, and the representative results of brain tissues are shown (A). Amounts of active/phospho-p38 were estimated by measuring the density of bands of phospho-p38 and expressed as arbitrary units (B). The data represent mean ± S.E. (n = 4–7). *, p < 0.02; **, p < 0.05. (C) mRNA was extracted from brain tissues from wild type and neuropathic Gaucher disease model V394L/PS-NA mice, and mRNAs of p38 isoforms and IL-6 were determined by the quantitative real time PCR. The data represent mean ± S.E. (n = 5). TNF-α, p < 0.0006 (*). (D) Serum IL-6 levels from wild type mice or Gaucher disease mouse models (V394L, D409H, and V394L/PS-NA) were determined by the ELISA system. The data represent mean ± S.E. Wild type, n = 21; V394L, n = 10; D409H, n = 7; V394L/PS-NA, n = 7. Wt vs V394L, p < 0.0006 (*); Wt vs D409, p < 0.03 (**); Wt vs V394L/PS-NA, p < 0.05 (***).

Mentions: To evaluate p38 activation in Gaucher’s disease in vivo, three types of Gaucher’s disease mouse models were employed, including V394L, D409H, and V394L/PS-NA. The former two types have been established as homozygous for GBA1 point mutant knock-in mice with minor glucosylceramide accumulation in viscera and without significant phenotypes such as neuro-degeneration and shorter life span [21]. The last has been generated by crossing V394L homozygous with the mouse with hypomorphic expression of the prosaposin transgenes (5–45% of wild type, PS-NA), showing glucosylceramide accumulation in multiple organs, several phenotypes including neuro-degeneration and shorter life span [22]. Phospho/active p38 in tissues was determined by immunoblotting. p38 in the brain tissues of neuronopathic mice (V394L/PS-NA) was activated as compared with those of wild type, whereas other Gaucher’s disease mice (V394L and D409H) did not show significant activation of p38 (Fig 1A and 1B). p38 activation in brain tissues is correlated with neuronopathic phenotype. As to other tissues including liver and lung, the levels of active p38 in all types of Gaucher’s disease mice were increased as compared with those of wild type (Fig 1B).


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 a Gaucher’s disease mouse model.Proteins were extracted from tissues (brain, lung, and liver) of wild type mice or Gaucher disease mouse models (V394L, D409H, and V394L/PS-NA) and then subjected to immunoblot analysis with antibodies specific for phospho- p38 and β-actin. Equal amounts of protein were loaded in each lane, and the representative results of brain tissues are shown (A). Amounts of active/phospho-p38 were estimated by measuring the density of bands of phospho-p38 and expressed as arbitrary units (B). The data represent mean ± S.E. (n = 4–7). *, p < 0.02; **, p < 0.05. (C) mRNA was extracted from brain tissues from wild type and neuropathic Gaucher disease model V394L/PS-NA mice, and mRNAs of p38 isoforms and IL-6 were determined by the quantitative real time PCR. The data represent mean ± S.E. (n = 5). TNF-α, p < 0.0006 (*). (D) Serum IL-6 levels from wild type mice or Gaucher disease mouse models (V394L, D409H, and V394L/PS-NA) were determined by the ELISA system. The data represent mean ± S.E. Wild type, n = 21; V394L, n = 10; D409H, n = 7; V394L/PS-NA, n = 7. Wt vs V394L, p < 0.0006 (*); Wt vs D409, p < 0.03 (**); Wt vs V394L/PS-NA, p < 0.05 (***).
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pone.0136633.g001: p38 activation and IL-6 formation in a Gaucher’s disease mouse model.Proteins were extracted from tissues (brain, lung, and liver) of wild type mice or Gaucher disease mouse models (V394L, D409H, and V394L/PS-NA) and then subjected to immunoblot analysis with antibodies specific for phospho- p38 and β-actin. Equal amounts of protein were loaded in each lane, and the representative results of brain tissues are shown (A). Amounts of active/phospho-p38 were estimated by measuring the density of bands of phospho-p38 and expressed as arbitrary units (B). The data represent mean ± S.E. (n = 4–7). *, p < 0.02; **, p < 0.05. (C) mRNA was extracted from brain tissues from wild type and neuropathic Gaucher disease model V394L/PS-NA mice, and mRNAs of p38 isoforms and IL-6 were determined by the quantitative real time PCR. The data represent mean ± S.E. (n = 5). TNF-α, p < 0.0006 (*). (D) Serum IL-6 levels from wild type mice or Gaucher disease mouse models (V394L, D409H, and V394L/PS-NA) were determined by the ELISA system. The data represent mean ± S.E. Wild type, n = 21; V394L, n = 10; D409H, n = 7; V394L/PS-NA, n = 7. Wt vs V394L, p < 0.0006 (*); Wt vs D409, p < 0.03 (**); Wt vs V394L/PS-NA, p < 0.05 (***).
Mentions: To evaluate p38 activation in Gaucher’s disease in vivo, three types of Gaucher’s disease mouse models were employed, including V394L, D409H, and V394L/PS-NA. The former two types have been established as homozygous for GBA1 point mutant knock-in mice with minor glucosylceramide accumulation in viscera and without significant phenotypes such as neuro-degeneration and shorter life span [21]. The last has been generated by crossing V394L homozygous with the mouse with hypomorphic expression of the prosaposin transgenes (5–45% of wild type, PS-NA), showing glucosylceramide accumulation in multiple organs, several phenotypes including neuro-degeneration and shorter life span [22]. Phospho/active p38 in tissues was determined by immunoblotting. p38 in the brain tissues of neuronopathic mice (V394L/PS-NA) was activated as compared with those of wild type, whereas other Gaucher’s disease mice (V394L and D409H) did not show significant activation of p38 (Fig 1A and 1B). p38 activation in brain tissues is correlated with neuronopathic phenotype. As to other tissues including liver and lung, the levels of active p38 in all types of Gaucher’s disease mice were increased as compared with those of wild type (Fig 1B).

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