Combination therapy with coenzyme Q10 and creatine produces additive neuroprotective effects in models of Parkinson's and Huntington's diseases - PubMed (original) (raw)

Combination therapy with coenzyme Q10 and creatine produces additive neuroprotective effects in models of Parkinson's and Huntington's diseases

Lichuan Yang et al. J Neurochem. 2009 Jun.

Abstract

Coenzyme Q(10) (CoQ(10)) and creatine are promising agents for neuroprotection in neurodegenerative diseases via their effects on improving mitochondrial function and cellular bioenergetics and their properties as antioxidants. We examined whether a combination of CoQ(10) with creatine can exert additive neuroprotective effects in a MPTP mouse model of Parkinson's disease, a 3-NP rat model of Huntington's disease (HD) and the R6/2 transgenic mouse model of HD. The combination of the two agents produced additive neuroprotective effects against dopamine depletion in the striatum and loss of tyrosine hydroxylase neurons in the substantia nigra pars compacta (SNpc) following chronic subcutaneous administration of MPTP. The combination treatment resulted in significant reduction in lipid peroxidation and pathologic alpha-synuclein accumulation in the SNpc neurons of the MPTP-treated mice. We also observed additive neuroprotective effects in reducing striatal lesion volumes produced by chronic subcutaneous administration of 3-NP to rats. The combination treatment showed significant effects on blocking 3-NP-induced impairment of glutathione homeostasis and reducing lipid peroxidation and DNA oxidative damage in the striatum. Lastly, the combination of CoQ(10) and creatine produced additive neuroprotective effects on improving motor performance and extending survival in the transgenic R6/2 HD mice. These findings suggest that combination therapy using CoQ(10) and creatine may be useful in the treatment of neurodegenerative diseases such as Parkinson's disease and HD.

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Figures

Fig. 1

Fig. 1. Additive neuroprotective effects of creatine with CoQ10 in attenuating MPTP-induced nigrostriatal dopaminergic neurodegeneration

Mice were fed with either an unsupplemented control diet or a diet supplemented with 2% creatine or 1% CoQ10 or a combination of two for one week before implanted subcutaneously with osmotic minipumps that delivered MPTP at a dose of 40mg/kg daily for 28 day. A. Striatal levels of dopamine analyzed by HPLC show chronic MPTP administration resulted in a significant depletion of striatal dopamine, and treatment with creatine, CoQ10 or their combination significantly attenuated MPTP-induced depletion of striatal dopamine, in which the combination treatment showed an additive protection effect. B. Stereologic cell counts of total (Nissl-positive) and TH-immunopositive dopaminergic neurons in the SNpc showed a significant cell loss following MPTP administration which was significantly attenuated by creatine, CoQ10 or their combination treatment. The combination exerted significantly better protection than either the creatine or the CoQ10. n = 15 mice per group. Data represent means ± SEM, * P < 0.05 ** P < 0.01 when compared with the control diets with MPTP group; #p< 0.05 compared to either creatine or CoQ10 diet with MPTP group, by ANOVA. C. Representative photomicrographs of TH-immunostained sections through the SNpc of mice show a significant reduction in TH-positive neurons of SNpc by chronic MPTP, and treatment of creatine, CoQ10 or their combination significantly blocked MPTP-induced loss of TH-positive neurons. In figure: a, control diet alone; b, control diet with MPTP; c, 2% creatine diet with MPTP; d, 1% CoQ10 with MPTP; e, the combination diet with MPTP.

Fig. 2

Fig. 2. Creatine, CoQ10 and their combination block chronic MPTP-induced lipid peroxidation and α-synuclein accumulation in SNpc dopaminergic neurons

A. Photomicrographs of MDA-immunostained sections through the SNpc of mice pretreated with control, 2% creatine, 1% CoQ10 or the combination of two for 7 days followed with a chronic administration of MPTP 40mg/kg daily for 28 days, show an increase in MDA staining in SNpc cells of mice with MPTP and control diet. Creatine, CoQ10 or their combination treatment resulted in a marked reduction of MPTP-induced MDA immunostaining. High magnification inserts show the MDA staining in neurons. B. Using sections from control mice with control diet as the baseline background, stereological analysis showed the total number of MDA-positive neurons in SNpc of MPTP-treated mice was significantly reduced by the treatment of creatine, CoQ10 or their combination. C. Photomicrographs of α-synuclein stained sections through the SNpc of mice described above show a significant increase of α-synuclein accumulation in SNpc of MPTP treated animals on control diet. Creatine, CoQ10 or their combination treatment resulted in a significant reduction of MPTP-induced α-synuclein accumulation in SNpc neurons. High magnification inserts show the α-synuclein staining in neurons. D. Quantification of α-synuclein labeled cells was performed by determining the average number of intensely labeled neurons per section, and only labeled neurons in MPTP-treated groups with staining intensity higher than that of controls were counted. Treatment with creatine, CoQ10 or their combination reduced α-synuclein positive cells in SNpc, with a significant reduction reached by combination treatment. In figure: a, control diet alone; b, control diet with MPTP; c, 2% creatine diet with MPTP; d, 1% CoQ10 with MPTP; e, the combination diet with MPTP. n= 5 mice in each group. Scale bar = 100μm. Data represent means ± SEM, * P < 0.05. E. Confocal photomicrographs of sections through the SNpc of mice on chronic MPTP stained with anti-TH antibody (green) and anti-α-synuclein antibody (red) show the colocalization of the α-synuclein in the TH-positive neurons (yellow in merge), using a Zeiss LSM510 confocal microscope. Scale bar = 100μm.

Fig. 3

Fig. 3. Additive neuroprotective effects of creatine with CoQ10 in reducing striatal damage caused by 3-nitropropionic toxicity

Lewis rats pretreated with either an unsupplemented control diet or a diet supplemented with 2% creatine or 1% CoQ10 or a combination of two for one week were subcutaneously delivered with 3-NP (50 mg/kg/day) or PBS by implanted osmotic pumps for 7 days. A. The measurement of lesion volume in coronal brain slices of 3-NP treated controls shows obvious striatal damage caused by 3-NP toxicity, which was significantly reduced by the treatment of creatine, CoQ10 or the combination diet. The combination exerted an additive protective effect in reducing the volume of 3-NP-induced striatal lesion. Data represent mean ± SEM. *p < 0.05, when compared to 3-NP with control diet; # p < 0.05 compared to 3-NP group with either creatine or CoQ10 diet, by ANOVA. n= 15 rats per group. B. Representative photomicrographs of NeuN-immunostained sections through the coronal section of striatum and cortex of rats as described above show a marked lesion area in striatum of 3-NP treated rats with control diet contrasted to the intact PBS treated controls. The treatment with creatine, CoQ10 or the combination diet significantly reduced the 3-NP-induced lesion area in the striatum, which was the smallest in the combination diet treatment section (upper panel). High magnifications in lower panel show the loss of the striatal NeuN-positive neurons caused by 3-NP toxicity and the preservation exerted by the treatment of creatine, CoQ10 or the combination diet. In figure: a, control diet alone; b, control diet with 3-NP; c, 2% creatine diet with 3-NP; d, 1% CoQ10 with 3-NP; e, the combination diet with 3-NP.

Fig. 4

Fig. 4. Creatine, CoQ10 or their combination blocks 3-NP-induced impairment of striatal glutathione homeostasis and reduces oxidative damage

The striatum and cortex tissues collected from the Lewis rats described in methods were used for analysis of glutathione metabolites and oxidative damage markers. A. 3-NP administration resulted in a statistically significant reduction of striatal levels of GSH and the ratio of GSH/GSSG, whereas levels of GSSG were not significantly affected by 3-NP treatment measured by HPLC. Creatine, CoQ10 or the combination diet treatment significantly blocked 3-NP induced reductions in striatal levels of GSH and the ratio GSH/GSSG without impacting the GSSG levels. The combination diet exerted an additive effect in preserving striatal GSH/GSSG ratio. B. HPLC analysis of MDA levels in striatum revealed a statistically significant increase in MDA levels in 3-NP intoxicated rats treated with control diet. Creatine, CoQ10 or the combination diet treatment significantly reduced 3-NP-induced MDA formation in the striatum with the combination exerting a more significant reduction in striatal MDA. C. HPLC analysis of the ratio of 8OH2dG over dG, a marker of DNA oxidation, showed a significant increase in oxidative DNA damage in the cerebral cortex following 3-NP treatment, which is attenuated by treatment of creatine, CoQ10 or the combination diet, with the combination treatment reaching a statistically significant attenuation. Data represent mean ± SEM. *p< 0.05, ** p< 0.01 compared to control alone; #p< 0.05, ##p< 0.01 compared to 3-NP with control diet; + p< 0.05 compared to 3-NP with either creatine or CoQ10 diet, by ANOVA. n= 15 rats per group.

Fig. 5

Fig. 5. Additive effect of creatine and CoQ10 on improving motor performance and extending survival of R6/2 HD mice

Oral administration of either an unsupplemented control diet or a diet supplemented with 2% creatine or 1% CoQ10 or a combination of these two was started in R6/2 HD mice at 21 days of age old. Motor performance of R6/2 HD mice and wild-type (WT) littermates was evaluated by recording twice weekly the time (up to 3 min) that they remained on a rotarod turning at 16 rpm (A). Significant differences were seen between R6/2 mice and WT littermates. Administration of CoQ10 or creatine in R6/2 HD mice significantly improved rotarod performance starting at 5 weeks through 13 weeks of age, in comparison to R6/2 HD mice treated with control diet, and the greatest improvement in rotarod occurred using the combined administration of creatine and CoQ10 (see results section for statistic values). Kaplan-Meier probability of survival analysis (B) and the column graph (C) show that creatine, CoQ10 or the combination diet significantly extended R6/2 HD mice survival compared to the R6/2 mice treated with control diet. The combination diet produced an additive effect in extending the survival. **p< 0.01 compared to control diet group; #p< 0.05 compared to either creatine or CoQ10 diet group. n= 20 mice per group.

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References

    1. Agarwal R, Chase SD. Rapid, fluorimetric-liquid chromatographic determination of malondialdehyde in biological samples. J Chromatogr B Analyt Technol Biomed Life Sci. 2002;775:121–126. - PubMed
    1. Andreassen OA, Dedeoglu A, Ferrante RJ, Jenkins BG, Ferrante KL, Thomas M, Friedlich A, Browne SE, Schilling G, Borchelt DR, Hersch SM, Ross CA, Beal MF. Creatine increase survival and delays motor symptoms in a transgenic animal model of Huntington’s disease. Neurobiol Dis. 2001;8:479–491. - PubMed
    1. Beal MF, Shults CW. Effects of Coenzyme Q10 in Huntington’s disease and early Parkinson’s disease. Biofactors. 2003;18:153–161. - PubMed
    1. Beal MF, Ferrante RJ. Experimental therapeutics in transgenic mouse models of Huntington’s disease. Nat Rev Neurosci. 2004;5:373–384. - PubMed
    1. Beal MF, Henshaw DR, Jenkins BG, Rosen BR, Schulz JB. Coenzyme Q10 and nicotinamide block striatal lesions produced by the mitochondrial toxin malonate. Ann Neurol. 1994;36:882–888. - PubMed

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