glutathione homocysteine aging

J Nutr. 2006 Jun;136(6 Suppl):1660S-1665S. Related Articles, Links
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The effects of sulfur amino acid intake on immune function in humans.

Grimble RF.

Institute of Human Nutrition, School of Medicine, University of Southampton, Southampton SO16 7PX, UK.

No direct data exist on the influence of supranormal intakes of sulfur amino acids on immune function in humans. However 3 major products of sulfur amino acids, glutathione (GSH), homocysteine (Hcy), and taurine (Tau), influence, mainly, inflammatory aspects of the immune response in vitro and in vivo. Methionine intakes above approximately 1 g/d transiently raise plasma Tau, Hcy, and GSH. Tau and GSH ameliorate inflammation. Hcy has the opposite effect. A biphasic relation, between cellular GSH and CD4+ and CD8+ numbers occurs in healthy men. How changes in sulfur amino acid intake influence this phenomenon is unknown. In animals, high Tau intakes are antiinflammatory. How immune function in humans is affected is unknown. A positive relation between plasma neopterin (a marker of a Th-1-type immune response) and Hcy indicates that Hcy may play a part in inflammatory aspects of Parkinson’s disease and aging. In vitro, Hcy, at concentrations seen following consumption of approximately 6 g L-methionine/d in adults, increases the interactions among T lymphocytes, monocytes, and endothelium. Whether a similar phenomenon occurs in vivo is unknown. Polymorphisms in the methylenetetrahydrofolate reductase gene are associated with raised plasma Hcy in young but not old subjects. The relation of this observation to immune function is unknown. The relationships among Hcy, inflammatory aspects of disease, and in vitro alterations in immune cell behavior create a cautionary note about supplementation of diets with l-methionine to raise intake above approximately 1 g/d. Studies directly linking methionine intake, genetics, plasma Hcy, Tau, and GSH and immune function are needed.

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PMID: 16702336 [PubMed – indexed for MEDLINE]

2: Mech Ageing Dev. 2006 May;127(5):444-50. Epub 2006 Mar 6. Related Articles, Links
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Methionine flux to transsulfuration is enhanced in the long living Ames dwarf mouse.

Uthus EO, Brown-Borg HM.

US Department of Agriculture, ARS, Grand Forks Human Nutrition Research Center, ND 58202-9034, USA. .

Long-lived Ames dwarf mice lack growth hormone, prolactin, and thyroid stimulating hormone. Additionally the dwarf mice have enzyme activities and levels that combat oxidative stress more efficiently than those of normal mice. We have shown that methionine metabolism in Ames mice is markedly different than in their wild type littermates. In our previous work we hypothesized that the flux of methionine to the transsulfuration pathway is enhanced in the dwarf mice. The current study was designed to determine whether the flux of methionine to the transsulfuration pathway is increased. We did this by injecting either l-[methyl-(3)H]-methionine or l-[(35)S]-methionine into dwarf or normal mice and then determined retained label (in form of S-adenosylmethionine) 45 min later. The amount of retained hepatic (3)H and (35)S label was significantly reduced in the dwarf mice; at 45 min the specific radioactivity of SAM (pCi/nmol SAM) was 56% lower (p < 0.05) for (3)H-label and 64% lower (p < 0.005) for (35)S-label in dwarf than wild type mice. Retention of (35)S was significantly lower in the brain (37%, p < .04) and kidney (47%, p < 0.02) of the dwarf compared to wild type mice; there was no statistical difference in retained (3)H-label in either brain or kidney. This suggests that both the methyl-moiety and the carbon chain of methionine are lost much faster in the dwarf compared to the wild type mouse, implying that both transmethylation in the liver and transsulfuration in the liver, brain, and kidney are increased in the dwarf mice. As further support, we determined by real-time RT PCR the expression of methionine metabolism genes in livers of mice. Compared to wild type, the Ames dwarf had increased expression of methionine adenosyltransferase 1a (2.3-fold, p = 0.013), glycine N-methyltransferase (3.8-fold, p = 0.023), betaine homocysteine methyltransferase (5.5-fold, p = 0.0006), S-adenosylhomocysteine hydrolase (3.8-fold, p = 0.0005), and cystathionase (2.6-fold; tended to be increased, p = 0.055). Methionine synthase expression was significantly decreased in dwarf compared to wild type (0.48-fold, p = 0.023). These results confirm that the flux of methionine to transsulfuration is enhanced in the Ames dwarf. This, along with data from previous studies support the hypothesis that altered methionine metabolism plays a significant role in the oxidative defense of the dwarf mouse and that the mechanism for the enhanced oxidative defense may be through altered GSH metabolism as a result of the distinctive methionine metabolism.

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PMID: 16519922 [PubMed – indexed for MEDLINE]

3: J Nutr. 2006 Feb;136(2):373-8. Related Articles, Links
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Plasma glutathione and cystathionine concentrations are elevated but cysteine flux is unchanged by dietary vitamin B-6 restriction in young men and women.

Davis SR, Quinlivan EP, Stacpoole PW, Gregory JF 3rd.

Food Science andHuman Nutrition Department, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, FL 32611-0370, USA.

Cysteine synthesis from homocysteine is catalyzed by two pyridoxal 5′-phosphate (PLP)-dependent enzymes. This suggests that vitamin B-6 status might affect cysteine and glutathione homeostasis, but it is unclear whether this occurs in humans. We assessed the effects of vitamin B-6 status on static and kinetic parameters of cysteine and glutathione metabolism in healthy female (n=5) and male (n=4) volunteers (20-30 y) before and after 4 wk of dietary vitamin B-6 restriction (<0.5 mg vitamin B-6/d). Rates of reactions related to cysteine metabolism were measured from blood sampled during primed, constant infusions of [(13)C(5)]methionine, [3-(13)C]serine, and [(2)H(2)]cysteine that were conducted after an overnight fast at baseline and after the dietary protocol. Vitamin B-6 restriction reduced the concentration of PLP (55.1+/- 8.3 vs. 22.6+/-1.3 nmol/L; P=0.004) and increased concentrations of cystathionine (124%; P<0.001) and total glutathione (38%; P<0.008) in plasma. Concentrations of plasma homocysteine, cysteine, cysteinylglycine, and C-reactive protein (an indicator of systemic inflammation) were not affected by dietary vitamin B-6 restriction. The rate of cysteine synthesis via transsulfuration was below detection limits in this protocol. Neither the fractional synthesis rate of cystathionine nor whole-body cysteine flux was affected by vitamin B-6 restriction. These data indicate that glutathione homeostasis is altered by dietary vitamin B-6 deficiency and appears to be unrelated to cysteine flux under conditions of minimal amino acid intake as evaluated in this study.

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PMID: 16424114 [PubMed – indexed for MEDLINE]

4: Exp Eye Res. 2004 Dec;79(6):875-86. Related Articles, Links
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The presence of a transsulfuration pathway in the lens: a new oxidative stress defense system.

Persa C, Pierce A, Ma Z, Kabil O, Lou MF.

Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln, NE 68583-0905, USA.

The finding that a lens under oxidative stress accumulated free and protein-bound cysteine (protein-S-S-cysteine) in the fiber cells prompted us to examine if there is an alternative source for cysteine pools besides the active cysteine transport system in the lens, namely, the transsulfuration pathway of homocysteine-cystathionine-cysteine, which utilises methionine through transmethylation. We examined the presence of the gene for cystathionine-beta-synthase (CBS), the rate limiting enzyme that converts homocysteine to cystathionine in the transsulfuration pathway, in human lens epithelial (HLE) B3 cells using PCR with primers designed based on the sequence of human liver CBS (Forward 5′-CCA CAC TGC CCC GGC AAA AT-3′; Reverse 5′-CTG GCA ATG CCC GTG ATG GT-3′). The purified DNA fragment (586 bp) from PCR analysis was sequenced and confirmed the homology with CBS gene from other human tissues. The CBS protein band (67 kDa) was present in the HLE cells, which reacted positively with the human liver anti-CBS antibody. The enzyme protein was detected in the pig and human lenses with the highest intensity in the epithelial layer, lower but equal quantities of CBS was present in the cortical and nuclear regions. Human nuclear CBS increased while epithelial CBS decreased with aging. Oxidative stress transiently upregulated the gene expression of CBS both in HLE cells (0.1 mMH2O2) and in pig lens cultured in TC 199 medium (0.5 mMH2O2). The catalytic activity for CBS, which was assayed by measuring the production of C14-cystathionine from C14-serine in the presence of homocysteine, S-adenosyl-methionine and pyridoxal phosphate, was detectable in the HLE cells and transiently activated with H2O2. Free cystathionine accumulated when HLE B3 cells were treated with propargylglycine (PGG), an inhibitor of cystathionase, the downstream enzyme that converts cystathionine to cysteine. More cystathionine accumulation occurred when the cells were simultaneously exposed to PGG and 0.1 mMH2O2. We have shown that oxidative stress of H2O2 could increase the flux of this transsulfuration pathway by committing more homocysteine to cysteine and glutathione production as H2O2 (0.1 mM) inhibited the remethylation enzyme of methionine synthase while concurrently activating the CBS enzyme. This is the first evidence that a transsulfuration pathway is present in the lens, and that it can be upregulated under oxidative stress to provide additional redox potential for the cells.

PMID: 15642325 [PubMed – indexed for MEDLINE]

5: J Lab Clin Med. 2004 Nov;144(5):235-45. Related Articles, Links
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The effects of age and hyperhomocysteinemia on the redox forms of plasma thiols.

Di Giuseppe D, Frosali S, Priora R, Di Simplicio FC, Buonocore G, Cellesi C, Capecchi PL, Pasini FL, Lazzerini PE, Jakubowski H, Di Simplicio P.

Department of Neuroscience, Pharmacology Unit, University of Sienna, Sienna, Italy.

We assayed the redox forms of cysteine (reduced [CSH], oxidized [CSSC], and bound to protein [CS-SP]), cysteinylglycine (CGSH; cysteinylgycine disulfide [CGSSGC] and cysteinylglycine-protein mixed disulfide [CGS-SP]), glutathione (GSH; glutathione disulfide [GSSG] and glutathione-protein mixed disulfide [GS-SP]), homocysteine (Hcy; homocystine [HcyS] and homocystine-protein mixed disulfides [bHcy]), and protein sulfhydryls in the plasma of healthy subjects (divided into 8 groups ranging in age from birth to 70 years) and patients with mild hyperhomocysteinemia associated with cardiovascular disease (heart-transplant patients) or vascular atherosclerosis, with or without renal failure. In healthy individuals, levels of disulfides and protein-mixed disulfides were more abundant than those of thiols, and those of protein-thiol mixed disulfides were higher than disulfides. Concentrations of CSH, GSH, and CGSH in the various groups had profiles characterized by a maximum over time. The concentration of Hcy was unchanged up to the age of 30 years, after which it increased. CSSC concentration increased gradually with age, whereas concentrations of the other disulfides were essentially unchanged. By contrast, the concentrations of all protein-thiol mixed disulfides, especially those with CSH, increased gradually with age. Ranks of distribution of the reduced forms changed with age (at birth, CSH > CGSH > GSH > Hcy; in 1- to 2-year-olds, CSH > GSH > CGSH > Hcy; and in 51- to 70-year-olds, CSH > CGSH = GSH > Hcy), whereas those of disulfides and protein-thiol mixed disulfides were substantially unchanged (in all age groups, CSSC > CGSSGC > GSSG = HcyS and CS-SP > CGS-SP > bHcy > GS-SP). In patients with pathologic conditions, plasma levels of disulfide forms CSSC, HcyS, CS-SP, and bHcy were significantly increased, whereas other redox forms of thiols were unchanged or showed variations opposite (increasing or decreasing) to control values. Maximal increases in disulfides and protein-thiol mixed disulfides were associated with renal failure. Our data suggest that increases in plasma bHcy concentrations in subjects with pathologic conditions were more likely the result of activation of thiol-disulfide exchange reactions between free reduced Hcy and CS-SP than of a direct action of reactive oxygen species.

PMID: 15570241 [PubMed – indexed for MEDLINE]

6: Acta Biochim Pol. 2004;51(3):815-24. Related Articles, Links
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Plasma levels of total, free and protein bound thiols as well as sulfane sulfur in different age groups of rats.

Iciek M, Chwatko G, Lorenc-Koci E, Bald E, Wlodek L.

Institute of Medical Biochemistry, Collegium Medicum, Jagiellonian University, Krakow, Poland.

The redox status of plasma thiols can be a diagnostic indicator of different pathological states. The aim of this study was to identify the age dependent changes in the plasma levels of total, free and protein bound glutathione, cysteine and homocysteine. The determination was conducted in plasma of three groups of rats: 1) young (3-month-old), 2) middle aged (19-month-old), and 3) old (31-month-old). Total levels of glutathione, cysteine and homocysteine and their respective free and protein-bound fractions decreased with age. The only exception was a rise in free homocysteine concentration in the middle group, which indicates a different pattern of transformations of this thiol in plasma. The drop in the level of protein-bound thiols suggests that the antioxidant capacity of plasma diminishes with age, which, consequently, leads to impaired protection of -SH groups through irreversible oxidation. The plasma sulfane sulfur level also declines with age, which means that aging is accompanied by inhibition of anaerobic sulfur metabolism.

PMID: 15448741 [PubMed – indexed for MEDLINE]

7: J Biol Inorg Chem. 2004 Apr;9(3):269-80. Epub 2004 Feb 3. Related Articles, Links
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Iron inhibits neurotoxicity induced by trace copper and biological reductants.

White AR, Barnham KJ, Huang X, Voltakis I, Beyreuther K, Masters CL, Cherny RA, Bush AI, Cappai R.

Department of Pathology and Centre for Neuroscience, The University of Melbourne, 3010, Carlton South, Victoria, Australia.

The extracellular microenvironment of the brain contains numerous biological redox agents, including ascorbate, glutathione, cysteine and homocysteine. During ischemia/reperfusion, aging or neurological disease, extracellular levels of reductants can increase dramatically owing to dysregulated homeostasis. The extracellular concentrations of transition metals such as copper and iron are also substantially elevated during aging and in some neurodegenerative disorders. Increases in the extracellular redox capacity can potentially generate neurotoxic free radicals from reduction of Cu(II) or Fe(III), resulting in neuronal cell death. To investigate this in vitro, the effects of extracellular reductants (ascorbate, glutathione, cysteine, homocysteine or methionine) on primary cortical neurons was examined. All redox agents except methionine induced widespread neuronal oxidative stress and subsequent cell death at concentrations occurring in normal conditions or during neurological insults. This neurotoxicity was totally dependent on trace Cu (>or=0.4 microM) already present in the culture medium and did not require addition of exogenous Cu. Toxicity involved generation of Cu(I) and H(2)O(2), while other trace metals did not induce toxicity. Surprisingly, administration of Fe(II) or Fe(III) (>or=2.5 microM) completely abrogated reductant-mediated neurotoxicity. The potent protective activity of Fe correlated with Fe inhibiting reductant-mediated Cu(I) and H(2)O(2) generation in cell-free assays and reduced cellular Cu uptake by neurons. This demonstrates a novel role for Fe in blocking Cu-mediated neurotoxicity in a high reducing environment. A possible pathogenic consequence for these phenomena was demonstrated by abrogation of Fe neuroprotection after pre-exposure of cultures to the Alzheimer’s amyloid beta peptide (Abeta). The loss of Fe neuroprotection against reductant toxicity was greater after treatment with human Abeta1-42 than with human Abeta1-40 or rodent Abeta1-42, consistent with the central role of Abeta1-42 in Alzheimer’s disease. These findings have important implications for trace biometal interactions and free radical-mediated damage during neurodegenerative illnesses such as Alzheimer’s disease and old-age dementia.

PMID: 14758525 [PubMed – indexed for MEDLINE]

8: J Pineal Res. 2002 Oct;33(3):151-5. Related Articles, Links
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Effects of pinealectomy on the levels and the circadian rhythm of plasma homocysteine in rats.

Baydas G, Gursu MF, Cikim G, Canpolat S, Yasar A, Canatan H, Kelestimur H.

Department of Physiology, College of Medicine, Firat University, Elazyg 23119, Turkey.

Hyperhomocysteinemia is an independent cardiovascular risk factor. There are several factors including aging that contribute to the development of hyperhomocysteinemia. Nevertheless, the exact mechanisms causing this condition are still debated. We hypothesize that the age-related decrease in melatonin levels may be consequential in hyperhomocysteinemia. Recently, we found that plasma homocysteine (Hcy) levels are increased in pinealectomized (PINX) rats and melatonin reverses this increase. The aim of the present study was to determine if there is a circadian rhythm of plasma Hcy in rats and to examine the effect of pinealectomy on this cycle. Plasma Hcy levels demonstrated a 24-hr rhythm with a peak at 02:00 hr and a nadir at 14:00 hr in both control and PINX rats. Pinealectomy did not change the phase of the rhythm or the nocturnal elevation of plasma Hcy, but it did significantly increase mean plasma Hcy levels compared with those in controls and in rats that were sham pinealectomized (sPINX) (P < 0.05). Melatonin decreases plasma Hcy levels while causing an increase in total glutathione (tGSH). In conclusion, we speculate that decreasing levels of melatonin during aging lead to hyperhomocysteinemia and a decrease in tGSH and the latter may be one of the factors causing hyperhomocysteinemia in the elderly population.

PMID: 12220329 [PubMed – indexed for MEDLINE]

9: J Clin Lab Anal. 2001;15(3):144-53. Related Articles, Links
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Simultaneous determination of total plasma glutathione, homocysteine, cysteinylglycine, and methionine by high-performance liquid chromatography with electrochemical detection.

Houze P, Gamra S, Madelaine I, Bousquet B, Gourmel B.

Laboratoire de Biochimie A, Hopital Saint-Louis, Paris, France.

We here describe an ion-exchange high-performance liquid chromatography technique with electrochemical detection for rapid quantification of glutathione, homocysteine, cysteinylglycine, and methionine. The analytical validation of the technique showed within-assay and between-assay coefficients of variation between 3.1 and 4.3%, and 3.7 and 8.6%, respectively. Percentages of recovery for overload and dilution tests were between 87 and 120%. Detection limits were 1 micromol/L for methionine and 0.5 micromol/L for other compounds. There was no interference with any physiological and pharmacological substances possessing a thiol function. Aminothiol concentrations determined in 100 control subjects (50 women and 50 men) showed no age- or sex-rated differences for except for homocysteine which was increased (+ 28%) in oldest subjects of both sexes. In 60 patients at risk (30 with chronic renal failure, 30 with diabetes), homocysteine concentration was significantly increased. No variation in other aminothiols was observed in diabetic subjects. Methionine was decreased and cysteinylglycine was increased in patients with chronic renal failure. The present technique-rapid, easy to use, and reliable-appears suitable for routine application in the exploration of aminothiol metabolic pathways including mechanisms of hyperhomocysteinemia.

PMID: 11344530 [PubMed – indexed for MEDLINE]

10: Life Sci. 2000 Aug 4;67(11):1317-24. Related Articles, Links

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