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Relative importance of redox buffers GSH and NAD(P)H in age-related neurodegeneration and Alzheimer disease-like mouse neurons.

Ghosh D, Levault KR, Brewer GJ - Aging Cell (2014)

Bottom Line: Further, although depletion of NAD(P)H or GSH correlated linearly with neuron death, compared with GSH depletion, higher neurodegeneration was observed when NAD(P)H was extrapolated to zero, especially in old age, and in the 3xTg-AD neurons.Our data indicate that in aging and more so in AD-like neurons, NAD(P)H redox control is upstream of GSH and an oxidative redox shift that promotes neurodegeneration.Thus, NAD(P)H generation may be a more efficacious therapeutic target upstream of GSH and ROS.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Microbiology, Immunology and Cell Biology, Southern Illinois University School of Medicine, Springfield, IL, 62794-9626, USA.

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Brain NADPH concentration and NADPH/NADP redox state decline with age and correlate with decline in brain expression of NAMPT and NNT gene expression. By HPLC analysis of cortical/hippocampal tissue homogenates from 4-, 11- and 21-month non-Tg (dashed line) and 3xTg-AD (solid line), A) NADPH concentration (nmol/mg) declines with age (ANOVA F(2,24) = 10.4, P = 0.001). B) Calculated NADPH/NADP redox state (mV) using the Nernst equation indicates a large oxidative shift in both non-Tg and 3xTg-AD brain with age (ANOVA F(2,24) = 15.7, P = 0.001). qRT PCR on non-Tg (gray filled circle, dashed line) and 3xTg-AD (black filled circle, solid line) brains indicate an age- and AD-related loss in gene expression of metabolic enzymes C) NAMPT (ANOVA, age F(3,33) = 21.8, P < 0.001, genotype F(1,33)=6.6, P = 0.02), and D) NNT (ANOVA age F(3,33) = 64.08, P < 0.001, genotype F(1,33) = 32.417, P < 0.001). Fold change expressed relative to 2 month non-Tg GAPDH as internal control. Brain NADPH levels correlate with E) NAMPT (non-Tg R2 = 0.76, 3xTg-AD R2 = 0.84) and F) NNT (non-Tg R2 = 0.76, 3xTg-AD R2 = 0.99). n = 4–7 animals per age per genotype. (G) Analysis of NAMPT protein by immunoblot indicates a larger decline with age than for mRNA and no genotype difference (n = 4 brains from each mouse).
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fig06: Brain NADPH concentration and NADPH/NADP redox state decline with age and correlate with decline in brain expression of NAMPT and NNT gene expression. By HPLC analysis of cortical/hippocampal tissue homogenates from 4-, 11- and 21-month non-Tg (dashed line) and 3xTg-AD (solid line), A) NADPH concentration (nmol/mg) declines with age (ANOVA F(2,24) = 10.4, P = 0.001). B) Calculated NADPH/NADP redox state (mV) using the Nernst equation indicates a large oxidative shift in both non-Tg and 3xTg-AD brain with age (ANOVA F(2,24) = 15.7, P = 0.001). qRT PCR on non-Tg (gray filled circle, dashed line) and 3xTg-AD (black filled circle, solid line) brains indicate an age- and AD-related loss in gene expression of metabolic enzymes C) NAMPT (ANOVA, age F(3,33) = 21.8, P < 0.001, genotype F(1,33)=6.6, P = 0.02), and D) NNT (ANOVA age F(3,33) = 64.08, P < 0.001, genotype F(1,33) = 32.417, P < 0.001). Fold change expressed relative to 2 month non-Tg GAPDH as internal control. Brain NADPH levels correlate with E) NAMPT (non-Tg R2 = 0.76, 3xTg-AD R2 = 0.84) and F) NNT (non-Tg R2 = 0.76, 3xTg-AD R2 = 0.99). n = 4–7 animals per age per genotype. (G) Analysis of NAMPT protein by immunoblot indicates a larger decline with age than for mRNA and no genotype difference (n = 4 brains from each mouse).

Mentions: These culture studies were complemented by in vivo measures of the NADPH/NADP redox state of brain tissue homogenates by HPLC from 4-, 11-, and 21-month non-Tg and 3xTg-AD animals. We observed that after middle age (11 month), the NADPH concentration declined similarly in both non-Tg and 3xTg-AD brain (Fig. 6A). Likewise, the redox state of both non-Tg and 3xTg-AD brain was significantly more oxidized with age (Fig. 6B) (ANOVA P = 0.001). However, we did not observe any genotype difference, possibly due to blood and glial compensation for neuronal changes. These results indicate that the redox state in the whole aging brain reflects an oxidative shift as seen in the cultured neurons.


Relative importance of redox buffers GSH and NAD(P)H in age-related neurodegeneration and Alzheimer disease-like mouse neurons.

Ghosh D, Levault KR, Brewer GJ - Aging Cell (2014)

Brain NADPH concentration and NADPH/NADP redox state decline with age and correlate with decline in brain expression of NAMPT and NNT gene expression. By HPLC analysis of cortical/hippocampal tissue homogenates from 4-, 11- and 21-month non-Tg (dashed line) and 3xTg-AD (solid line), A) NADPH concentration (nmol/mg) declines with age (ANOVA F(2,24) = 10.4, P = 0.001). B) Calculated NADPH/NADP redox state (mV) using the Nernst equation indicates a large oxidative shift in both non-Tg and 3xTg-AD brain with age (ANOVA F(2,24) = 15.7, P = 0.001). qRT PCR on non-Tg (gray filled circle, dashed line) and 3xTg-AD (black filled circle, solid line) brains indicate an age- and AD-related loss in gene expression of metabolic enzymes C) NAMPT (ANOVA, age F(3,33) = 21.8, P < 0.001, genotype F(1,33)=6.6, P = 0.02), and D) NNT (ANOVA age F(3,33) = 64.08, P < 0.001, genotype F(1,33) = 32.417, P < 0.001). Fold change expressed relative to 2 month non-Tg GAPDH as internal control. Brain NADPH levels correlate with E) NAMPT (non-Tg R2 = 0.76, 3xTg-AD R2 = 0.84) and F) NNT (non-Tg R2 = 0.76, 3xTg-AD R2 = 0.99). n = 4–7 animals per age per genotype. (G) Analysis of NAMPT protein by immunoblot indicates a larger decline with age than for mRNA and no genotype difference (n = 4 brains from each mouse).
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fig06: Brain NADPH concentration and NADPH/NADP redox state decline with age and correlate with decline in brain expression of NAMPT and NNT gene expression. By HPLC analysis of cortical/hippocampal tissue homogenates from 4-, 11- and 21-month non-Tg (dashed line) and 3xTg-AD (solid line), A) NADPH concentration (nmol/mg) declines with age (ANOVA F(2,24) = 10.4, P = 0.001). B) Calculated NADPH/NADP redox state (mV) using the Nernst equation indicates a large oxidative shift in both non-Tg and 3xTg-AD brain with age (ANOVA F(2,24) = 15.7, P = 0.001). qRT PCR on non-Tg (gray filled circle, dashed line) and 3xTg-AD (black filled circle, solid line) brains indicate an age- and AD-related loss in gene expression of metabolic enzymes C) NAMPT (ANOVA, age F(3,33) = 21.8, P < 0.001, genotype F(1,33)=6.6, P = 0.02), and D) NNT (ANOVA age F(3,33) = 64.08, P < 0.001, genotype F(1,33) = 32.417, P < 0.001). Fold change expressed relative to 2 month non-Tg GAPDH as internal control. Brain NADPH levels correlate with E) NAMPT (non-Tg R2 = 0.76, 3xTg-AD R2 = 0.84) and F) NNT (non-Tg R2 = 0.76, 3xTg-AD R2 = 0.99). n = 4–7 animals per age per genotype. (G) Analysis of NAMPT protein by immunoblot indicates a larger decline with age than for mRNA and no genotype difference (n = 4 brains from each mouse).
Mentions: These culture studies were complemented by in vivo measures of the NADPH/NADP redox state of brain tissue homogenates by HPLC from 4-, 11-, and 21-month non-Tg and 3xTg-AD animals. We observed that after middle age (11 month), the NADPH concentration declined similarly in both non-Tg and 3xTg-AD brain (Fig. 6A). Likewise, the redox state of both non-Tg and 3xTg-AD brain was significantly more oxidized with age (Fig. 6B) (ANOVA P = 0.001). However, we did not observe any genotype difference, possibly due to blood and glial compensation for neuronal changes. These results indicate that the redox state in the whole aging brain reflects an oxidative shift as seen in the cultured neurons.

Bottom Line: Further, although depletion of NAD(P)H or GSH correlated linearly with neuron death, compared with GSH depletion, higher neurodegeneration was observed when NAD(P)H was extrapolated to zero, especially in old age, and in the 3xTg-AD neurons.Our data indicate that in aging and more so in AD-like neurons, NAD(P)H redox control is upstream of GSH and an oxidative redox shift that promotes neurodegeneration.Thus, NAD(P)H generation may be a more efficacious therapeutic target upstream of GSH and ROS.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Microbiology, Immunology and Cell Biology, Southern Illinois University School of Medicine, Springfield, IL, 62794-9626, USA.

Show MeSH
Related in: MedlinePlus