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Glutathione & Whey Proteins in
Stroke, Coronary, Cardiovascular, Heart Disease

Bioactive Components of Whey and Cardiovascular Health (pdf)
Sharon K. Gerdes, Dr. W. James Harper, Ph.D, Dr. G. Miller, Ph.D.
(A Monograph Published by U.S. Dairy Export Council®)

Lowering effect of dietary milk-whey protein v. casein on plasma and liver cholesterol concentrations in rats
Zhang X, Beynen AC [Br J Nutr (1993 Jul) 70(1):139-46] The effect of dietary whey protein versus casein on plasma and liver cholesterol concentrations was investigated in female, weanling rats. Balanced, purified diets containing either whey protein or casein, or the amino acid mixtures simulating these proteins, were used. At the low dietary protein level, whey protein versus casein did not affect plasma total cholesterol, but lowered the concentration of liver cholesterol. At the high dietary-protein level, whey protein significantly lowered plasma and liver cholesterol and also plasma triacylglycerols. The hypocholesterolemic effect of whey protein was associated with a decrease in very-low-density-lipoprotein cholesterol. At the high dietary protein concentration, whey protein reduced the fecal excretion of bile acids when compared with casein. The effects of intact whey protein versus casein were not reproduced by the amino acid mixtures simulating these proteins. It is suggested tentatively that the cholesterol-lowering effect of whey protein in rats is caused by inhibition of hepatic cholesterol synthesis.

Serum Glutathione in Adolescent Males Predicts Parental Coronary Heart Disease
John A. Morrison, PhD; Donald W. Jacobsen, PhD; Dennis L. Sprecher, MD; Killian Robinson, MD; Philip Khoury, MS; Stephen R. Daniels, MD, PhD [Circulation. 1999;100:2244] Traditional risk factors account for only half of the morbidity and mortality from coronary heart disease (CHD). There is substantial evidence that oxidative injury plays a major role in the atherosclerotic process. Thus, antioxidants may protect against development of atherosclerosis. Glutathione, an intracellular tripeptide with antioxidant properties, may be protective. This study found that low tGSH in adolescent boys is a significant independent predictor of parental CHD, in addition to elevated LDL cholesterol, low HDL cholesterol, and elevated total serum homocysteine concentrations.

Oxidant stress in the vasculature
Maytin M, Leopold J, Loscalzo J. [Curr Atheroscler Rep 1999 Sep;1(2):156-64] Vascular disease and vasomotor responses are largely influenced by oxidant stress. Numerous cellular antioxidant systems exist to defend against oxidant stress; glutathione and the enzymes superoxide dismutase and glutathione peroxidase are critical for maintaining the redox balance of the cell. However, the redox state is disrupted by certain vascular diseases. It appears that oxidant stress both promotes and is induced by diseases such as hypertension, atherosclerosis, and restenosis as well as by certain risk factors for coronary artery disease including hyperlipidemia, diabetes, and cigarette smoking. Once oxidant stress is invoked, characteristic pathophysiologic features ensue, namely adverse vessel reactivity, vascular smooth muscle cell proliferation, macrophage adhesion, platelet activation, and lipid peroxidation.

Erythrocyte susceptibility to lipid peroxidation in patients with coronary atherosclerosis
Dincer Y, Akcay T, and others. [Acta Med Okayama 1999 Dec;53(6):259-64.] "Erythrocyte TBARS production was significantly higher in patients with coronary atherosclerosis than in the controls. On the other hand, the levels of plasma high-density lipoproteins, vitamin C, vitamin E and erythrocyte GSH were significantly lower, and the levels of plasma total cholesterol, triglycerides, low-density lipoproteins and TBARS were significantly higher in the patients with coronary atherosclerosis than in the controls. In conclusion, our results indicate that erythrocytes from patients with coronary atherosclerosis are more susceptible to oxidation than those of controls and that these patients have lowered antioxidant capacity as revealed by decreased plasma levels of vitamins C and E."

L-2-oxothiazolidine-4- carboxylic acid reverses endothelial dysfunction in patients with coronary artery disease
Vita JA, Frei B, and others. [J Clin Invest 1998 Mar 15;101(6):1408-14.] "Cellular redox state...is a potential target for therapy in patients with coronary artery disease." These data suggest that augmenting cellular glutathione levels improves EDNO action in human atherosclerosis. Cellular redox state may be an important regulator of EDNO action, and is a potential target for therapy in patients with coronary artery disease.

Glutathione infusion enhances coronary blood flow during oxidative stress
[Circulation 1998;97:2299-2301] Oxygen free radicals cause endothelial vasomotor dysfunction. Investigators found that reduced glutathione intravenously suppresses constriction of human coronary arteries in response to acetylcholine and increases the effect of nitroglycerin. Glutathione administration may be useful in patients with coronary artery disease, both as a result of improvement of endothelial dysfunction and augmentation of nitroglycerin-induced vasodilation and antiplatelet activity.

Serum glutathione in adolescent males predicts parental coronary heart disease

Morrison JA, Jacobsen DW, Sprecher DL, Robinson K, Khoury P, Daniels SR. [Circulation 1999 Nov 30;100(22):2244-7] There is substantial evidence that oxidative injury plays a major role in the atherosclerotic process. Thus, antioxidants may protect against development of atherosclerosis. Glutathione, an intracellular tripeptide with antioxidant properties, may be protective. This case-control study compared total serum glutathione (tGSH) in 81 adolescent male offspring of parents with premature CHD (ie, before 56 years of age) and 78 control male offspring of parents without known or suspected CHD. Case offspring had significantly lower tGSH than control offspring. Low tGSH in adolescent boys is a significant independent predictor of parental CHD, in addition to elevated LDL cholesterol, low HDL cholesterol, and elevated total serum homocysteine concentrations.

Glutathione reverses endothelial dysfunction and improves nitric oxide bioavailability
Prasad A, Andrews NP, Padder FA, Husain M, Quyyumi AA. [J Am Coll Cardiol 1999 Aug;34(2):507-14] We investigated whether glutathione (GSH), a reduced thiol that modulates redox state and forms adducts of nitric oxide (NO), improves endothelium-dependent vasomotion and NO activity in atherosclerosis. Endothelial dysfunction and reduced NO activity are associated with atherosclerosis and its clinical manifestations such as unstable angina. Thiol supplementation with GSH selectively improves human endothelial dysfunction by enhancing NO activity.

Macrophage foam cell formation during early atherogenesis is determined by the balance between pro-oxidants and anti-oxidants in arterial cells and blood lipoproteins
Aviram M. [Antioxid Redox Signal 1999 Winter;1(4):585-94] Atherosclerosis is a multifactorial disease, where more than one mechanism, along more than one step, contributes to macrophage cholesterol accumulation and foam cell formation, the hallmark of early atherogenesis. Intervention to inhibit LDL oxidation can affect the above additional LDL modifications. The balance between pro-oxidants and anti-oxidants in the LDL particle, as well as in arterial wall macrophages (such as NADPH oxidase vs. glutathione), determines the extent of LDL oxidation. Antioxidants can protect LDL from oxidation not only by their binding to the lipoprotein, but also following their accumulation in cells of the arterial wall.....the combination of antioxidants together with active paraoxonase decreases the formation of Ox-LDL and preserves PON1's ability to hydrolyze this atherogenic lipoprotein and hence, to attenuate atherosclerosis.

Hyperglycemia in diabetic rats reduces the glutathione content in the aortic tissue
Tachi Y, Okuda Y, Bannai C, Bannai S, Shinohara M, Shimpuku H, Yamashita K, Ohura K. [Life Sci 2001 Jul 20;69(9):1039-47] The glutathione redox cycle plays a major role in scavenging hydrogen peroxide (H2O2) under physiological conditions. Recently, we demonstrated that a high glucose concentration in the culture medium reduced the level of H2O2 scavenging activity of human vascular smooth muscle cells (hVSMCs). We also showed that a high glucose concentration reduced the intracellular glutathione (GSH) content and the rate of uptake of cystine, which itself is a rate-limiting factor that maintains the GSH level. In the present study, we investigated whether the hyperglycemic condition in diabetic rats impairs the glutathione content in the aortic tissue in vivo. We demonstrated in vivo that the hyperglycemic condition in STZ-induced diabetic Wistar rats and OLETF rats reduced the GSH content in aortic tissue. This suggested reduced glutathione redox cycle function of aorta.

Effect of administration of fermented milk containing whey protein concentrate to rats and healthy men on serum lipids and blood pressure
Kawase M, Hashimoto H, Hosoda M, Morita H, Hosono A. [J Dairy Sci 2000 Feb;83(2):255-63] The effect of fermented milk supplemented with whey protein concentrate on the serum lipid level of rats was investigated. The serum total cholesterol level for the group fed fermented milk was significantly lower than that of the control group in rats. After 8 wk, the high density lipoprotein cholesterol level for the fermented milk group showed a significant rise after 4 wk , whereas that of the placebo group showed no change even after 4 wk. The triglyceride level for the fermented milk group lowered significantly after 4 wk, whereas that of the placebo group showed no change even after 4 wk. The atherogenic index [(total cholesterol - high density lipoprotein cholesterol)/high-density lipoprotein cholesterol] for the fermented milk group decreased significantly from 4.24 to 3.52. The systolic blood pressure lowered significantly by the intake of fermented milk. On the other hand, such effect was not observed in the placebo group. These results indicate potential of the development of fermented milk with multiple therapeutic effects.

Effect of milk protein and fat intake on blood pressure and the incidence of cerebrovascular diseases in stroke-prone spontaneously hypertensive rats (SHRSP)
Ikeda K, Mochizuki S, Nara Y, Horie R, Yamori Y.
[J Nutr Sci Vitaminol (Tokyo) 1987 Feb;33(1):31-6] The intake of two milk protein-rich diets containing casein and whey protein attenuated the development of severe hypertension in stroke-prone spontaneously hypertensive rats (SHRSP), and extended their life span in comparison with SHRSP on a regular stock diet. Milk fat-rich diet intake reduced the incidence of cerebrovascular disease in SHRSP without a significant fall in blood pressure. These results suggest that certain milk components have a preventive effect on hypertension and cerebrovascular disease in SHRSP.

Angiotensin I-converting enzyme inhibitory properties of whey protein digests: concentration and characterization of active peptides
Pihlanto-Leppala A, Koskinen P, Piilola K, Tupasela T, Korhonen H
. [J Dairy Res 2000 Feb;67(1):53-64] The aim of this study was to identify whey-derived peptides with angiotensin I-converting enzyme (ACE) inhibitory activity. The bovine whey proteins alpha-lactalbumin and beta-lactoglobulin were hydrolysed with pepsin, trypsin, chymotrypsin, pancreatin, elastase or carboxypeptidase alone and in combination. Whey protein digests gave a 50% inhibition (IC50) of ACE activity at concentration ranges within 345-1733 micrograms/ml. The IC50 values for the 1-30 kDa fractions ranged from 485 to 1134 micrograms/ml and for the < 1 kDa fraction from 109 to 837 mg/ml. Several ACE-inhibitory peptides were isolated from the hydrolysates by reversed-phase chromatography, and the potencies of the purified peptide fractions had IC50 values of 77-1062 microM. The ACE-inhibitory peptides identified were alpha-lactalbumin fractions (50-52), (99-108) and (104-108) and beta-lactoglobulin fractions (22-25), (32-40), (81-83), (94-100), (106-111) and (142-146).

Lactokinins: whey protein-derived ACE inhibitory peptides
FitzGerald RJ, Meisel H.
[Nahrung 1999 Jun;43(3):165-7] Angiotensin-I-converting enzyme (ACE) has been classically associated with the renin-angiotensin system which regulates peripheral blood pressure. Peptides derived from the major whey proteins, i.e. alpha-lactalbumin (alpha-la) and beta-lactoglobulin (beta-lg) in addition to bovine serum albumin (BSA), inhibit ACE. While they do not have the inhibitory potency of synthetic drugs commonly used in the treatment of hypertension, these naturally occurring peptides may represent nutraceutical /functional food ingredients for the prevention/treatment of high blood pressure. Studies with gastric and pancreatic proteinase digests of whey proteins indicate that enzyme specificity rather than extent of hydrolysis dictates the ACE inhibitory potency of whey hydrolysates.

Glutathione deficiencies exacerbate response to stroke
Phyllis G. Paterson, University of Saskatchewan - Saskatoon. Canada [Invited Symposium: The Therapeutic Potential of Phase II Enzyme Induction] The cascade of events responsible for the death of neural cells following a stroke include depletion of ATP, glutamate excitotoxicity, calcium overload, and production of strong oxidants that can overwhelm antioxidant defense. Glutathione (GSH) has a central role within the finely tuned network of antioxidant systems that can respond to the oxidative insult through its functions in peroxide scavenging via glutathione-S-transferase and the family of glutathione peroxidases, regeneration of alpha-tocopherol, and inhibition of NFkB which is required for the expression of pro-inflammatory genes. Our laboratory has used a nutritional approach to study the effects of GSH depletion in a rat model of stroke. We have found that a deficiency of sulfur amino acids used as a model of reduced cysteine supply for synthesis depresses GSH concentration in a number of brain regions. A second study demonstrated that acute sulfur amino acid deficiency exacerbates brain damage in a rat model of global hemispheric hypoxic ischemia. Approaches described in the literature for maintaining GSH under ischemia conditions have also been targeted towards increasing its synthesis. Administering a GSH ester immediately after an ischemic insult offered neuroprotection in one study as did the delivery of N-acetylcysteine, a compound that supports GSH synthesis by acting as a cysteine precursor. The series of studies reviewed suggests that GSH is an important determinant of the extent of secondary tissue damage in animal models of stroke. Strategies to enhance GSH in brain should be tested for their therapeutic efficacy in the human condition of stroke. (Funded by the Heart and Stroke Foundation of Saskatchewan)

Milk whey protein decreases oxygen free radical production in a murine model of chronic iron-overload cardiomyopathy
Bartfay WJ, Davis MT, Medves JM, Lugowski S. [Can J Cardiol. 2003 Sep;19(10):1163-8.]
Chronic iron overload is a major cause of organ failure worldwide, but its pathogenesis remains to be elucidated. Mice receiving iron treatments with whey supplementation had significantly lower concentrations of cytotoxic aldehydes and significantly higher cardiac levels of GPx and GSH activity than did iron-only treated mice. Additional basic research is warranted to examine the exact mechanisms by which milk whey protein protects the heart.


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