Gordon M. Lowe

Garlic and Cardiovascular Disease: A Critical Review

Epidemiologic studies show an inverse correlation between garlic consumption and progression of cardiovascular disease. Cardiovascular disease is associated with multiple factors such as raised serum total cholesterol, raised LDL and an increase in LDL oxidation, increased platelet aggregation, hypertension, and smoking. Numerous in vitro studies have confirmed the ability of garlic to reduce these parameters. Thus, garlic has been shown to inhibit enzymes involved in lipid synthesis, decrease platelet aggregation, prevent lipid peroxidation of oxidized erythrocytes and LDL, increase antioxidant status, and inhibit angiotension-converting enzyme. These findings have also been addressed in clinical trials. The studies point to the fact that garlic reduces cholesterol, inhibits platelet aggregation, reduces blood pressure, and increases antioxidant status. Since 1993, 44% of clinical trials have indicated a reduction in total cholesterol, and the most profound effect has been observed in garlics ability to reduce the ability of platelets to aggregate. Mixed results have been obtained in the area of blood pressure and oxidative-stress reduction. The findings are limited because very few trials have addressed these issues. The negative results obtained in some clinical trials may also have resulted from usage of different garlic preparations, unknown active constituents and their bioavalability, inadequate randomization, selection of inappropriate subjects, and short duration of trials. This review analyzes in vitro and in vivo studies published since 1993 and concludes that although garlic appears to hold promise in reducing parameters associated with cardiovascular disease, more in-depth and appropriate studies are required.

Lycopene and β-carotene protect against oxidative damage in HT29 cells at low concentrations but rapidly lose this capacity at higher doses

Epidemiological studies have clearly demonstrated a link between dietary carotenoids and the reduced incidence of certain diseases, including some cancers. However recent intervention studies (e.g. ATBC, CARET and others) have shown that beta-carotene supplementation has little or no beneficial effect and may, in fact, increase the incidence of lung cancers in smokers. This presents a serious dilemma for the scientific community - are carotenoids at high concentrations actually harmful in certain circumstances? Currently, a significant number of intervention studies are on-going throughout the world involving carotenoids (of both natural and synthetic origin). Our approach has been to study the ability of supplementary carotenoids in protecting cells against oxidatively-induced DNA damage (as measured by the comet assay), and membrane integrity (as measured by ethidium bromide uptake). Both lycopene and beta-carotene only afforded protection against DNA damage (induced by xanthine/xanthine oxidase) at relatively low concentrations (1-3 microM). These levels are comparable with those seen in the plasma of individuals who consume a carotenoid-rich diet. However, at higher concentrations (4-10 microM), the ability to protect the cell against such oxidative damage was rapidly lost and, indeed, the presence of carotenoids may actually serve to increase the extent of DNA damage. Similar data were obtained when protection against membrane damage was studied. This would suggest that supplementation with individual carotenoids to significantly elevate blood and tissue levels is of little benefit and, may, in fact, be deleterious. This in vitro data presented maybe significant in the light of recent intervention trials.

Antioxidant properties of aged garlic extract: an in vitro study incorporating human low density lipoprotein.

Oxidation of low-density lipoprotein (LDL) has been recognized as playing an important role in the development and progression of atherosclerotic heart disease. Human LDL was isolated and challenged with a range of oxidants either in the presence or absence of AGE or its diethyl ether extract. Oxidative modification of the LDL fraction using CuSO(4), 5-lipoxygenase and xanthine/xanthine oxidase was monitored by both the appearance of thiobarbituric-acid substances (TBA-RS) and an increase in electrophoretic mobility. This study indicates that AGE is an effective antioxidant as it scavenged superoxide ions and reduced lipid peroxide formation in cell free assays. Superoxide production was completely inhibited in the presence of a 10% (v/v) aqueous preparation of AGE and reduced by 34% in the presence of a 10% (v/v) diethyl ether extract of AGE. The presence of 10% (v/v) diethyl ether extract of AGE significantly reduced Cu(2+) and 15-lipoxygenase-mediated lipid peroxidation of isolated LDL by 81% and 37%, respectively. In addition, it was found that AGE also had the capacity to chelate copper ions. In contrast, the diethyl ether extract of AGE displayed no copper binding capacity, but demonstrated distinct antioxidant properties. These results support the view that AGE inhibits the in vitro oxidation of isolated LDL by scavenging superoxide and inhibiting the formation of lipid peroxides. AGE was also shown to reduce LDL oxidation by the chelation of Cu(2+). Thus, AGE may have a role to play in preventing the development and progression of atherosclerotic disease.

Lycopene and Its Antioxidant Role in the Prevention of Cardiovascular Diseases—A Critical Review

The present review is based mainly on papers published between 2000 and 2011 and gives information about the properties of the carotenoid lycopene in chemical and biological systems and its possible role in preventing cardiovascular diseases (CVD). The main aim of this report is to highlight its role as an antioxidant, also reported are bioactive properties that may influence the development of foam cells and protection against endothelial cell damage. The paper will also examine recent observations that lycopene may improve blood flow and reduce inflammatory responses. Lycopene possesses antioxidant properties in vitro, and some epidemiological studies have reported protective effects against the progression of CVD. The oxidation of human low density lipoproteins (LDL) is a fundamental mechanism in the initiation of atherosclerosis. A beneficial role of lycopene as antioxidant in the prevention of CVD is suggested but the data are still controversial. Lycopene is believed to be the most potent carotenoid antioxidant in vitro. Tissue culture experiments and animal studies support potential cardioprotective effects for lycopene and other carotenoids in the blood. Most studies showed beneficial effects of lycopene to individuals who are antioxidant-deficient like elderly patients, or humans exposed to higher levels of oxidative stress like smokers, diabetics, hemodialysis patients and acute myocardial infarction patients. By defining the right population and combining antioxidant potentials of lycopene with vitamins and other bioactive plant compounds, the beneficial role of lycopene in CVD can be clarified in future studies.

Carotenoid Composition and Antioxidant Potential in Subfractions of Human Low-Density Lipoprotein

Carotenoids and vitamin E are transported in human plasma complexed with lipoproteins. The bulk of them are associated with low-density lipoprotein (LDL), in which form they may act as antioxidants and thus delay the onset of atherosclerosis. We used a simple, rapid, ultracentrifugation technique to fractionate plasma lipoproteins in self-generating gradients of iodixanol (Optiprep™), a non-ionic iodinated density gradient medium. The carotenoid content and composition of a number of LDL subfractions was determined by reversed-phase high-performance liquid chromatography. Lycopene, β-carotene and β-cryptoxanthin were mainly located in the larger, less-dense LDL particles whereas lutein and zeaxanthin were found preferentially in the smaller, more dense LDL particles. When the antioxidant content of these fractions was expressed per milligram of LDL protein, significantly lower concentrations of carotenoid and vitamin E were found to be associated with the smaller, protein-rich fractions of LDL. Strong positive correlations were found between total carotenoid and vitamin E plasma concentrations and the lag-time of Cu2+-mediated oxidation of LDL subfractions. The more dense LDL subfractions, which had lower levels of these antioxidants, were more readily oxidized, highlighting their possible role in atherosclerotic events.

Aged Garlic Extract and Its Constituents Inhibit Platelet Aggregation through Multiple Mechanisms

Aged garlic extract (AGE) is a complex mixture. Its constituents include allin, cycloalliin, S-allyl-L-cysteine, S-methyl-L-cysteine, S-ethylcysteine, S-1-proponyl-L-cysteine, S-allylmercapto-L-cysteine, fructosyl-arginine, and beta-chlorogenin. It also contains L-arginine, L-cysteine, and L-methionine. AGE reduces the parameters associated with cardiovascular disease, including the inhibition of platelet aggregation. However, the underlying mechanisms have yet to be established and are the subject of this study. AGE inhibited platelet aggregation in a concentration-dependent manner, and this achieved significance between concentrations of 1.56-25% (v:v). The constituents of AGE, when tested as a mixture, displayed a biphasic pattern of inhibition, although this was not significant. L-Methionine displayed anti-aggregatory properties at concentrations of 0.00078, 0.00625, and 0.1 mmol/L, whereas L-arginine and L-cysteine were effective at 9 mmol/L. However, when the constituents and the amino acids were tested together, significant inhibition of platelet aggregation was observed only at a concentration of 1 mmol/L. AGE also displayed disaggregatory properties at concentrations of 12.5 and 25% (v:v). The constituents and the amino acids, when tested as a mixture, displayed disaggregatory properties at concentrations of 0.25 and 1 mmol/L. In contrast, a diethyl-ether extract of AGE had no effect on platelet aggregation. When platelets were stimulated with either A23187 or ADP, an increase in intraplatelet Ca2+ accompanied platelet aggregation. This increase in Ca2+ was abolished in the presence of AGE. It is likely that AGE works in a synergistic manner and exerts multiple effects on the biochemical pathways involved in platelet aggregation.

Circulating ghrelin exists in both lipoprotein bound and free forms.

Introduction Ghrelin is a gastric peptide that has been implicated in the development of obesity and cardiovascular disease. It has been reported that ghrelin binds to lipoproteins, although the different binding patterns of acylated ghrelin (AG) and unacylated ghrelin (UAG) are still to be determined. Methods Lipoprotein fractions were generated using a self-generating iodixanol gradient. AG and UAG were measured using specific enzyme immunoassays. Results AG bound to all lipoproteins in approximately equal concentrations (VLDL 26%, LDL 22%, HDL 23%) and was present as a plasma protein (27%). UAG bound more specifically to HDL (49%) and was present as a plasma protein (48%). Conclusions The different binding patterns of AG and UAG may have significant implications for their biological effects, including roles in energy metabolism, the development of obesity and potentially in the modulation of cardiovascular disease.

Aged garlic extract inhibits platelet activation by increasing intracellular cAMP and reducing the interaction of GPIIb/IIIa receptor with fibrinogen.

AIMS Increased platelet aggregation plays an important role in the etiology of cardiovascular disease. Garlic inhibits platelet aggregation; however, the mechanisms involved have not clearly been defined. This study was undertaken to investigate the mechanisms by which an aged garlic extract (AGE) inhibits both the activation and aggregation of human platelets. MAIN METHODS Isolated human platelets were stimulated with ADP and their adhesion to fibrinogen was assessed using Rose Bengal or (51)Cr uptake. Activation of platelets was assessed using fluorescence activated cell sorting (FACS) analysis along with measurement of intracellular cAMP. KEY FINDINGS AGE at concentrations in the range of 3.12 to 12.5% (v/v) inhibited the binding of platelets to fibrinogen by approximately 40% when compared to control values in the Rose Bengal assay (P<0.05). In the (51)Cr experiments AGE significantly inhibited the binding of ADP-activated platelets to immobilized fibrinogen by 61.5% at 1.56% and 6.25% (v/v) of AGE respectively. At a concentration of 12.5% (v/v) the inhibition was 70.4% and at 25% (v/v) it was 64.5% respectively (P<0.05). In the fluorescence activated cell sorting (FACS) analysis, AGE significantly decreased the amount of PAC-1 binding to GPIIb/IIIa by approximately 72% compared with PBS control. In conjunction to these observations, AGE also increased platelet cAMP (P<0.01) levels. SIGNIFICANCE These findings suggest that AGE inhibits platelet aggregation via inhibition of the GPIIb/IIIa receptor and an increase in cAMP.

Characterisation of a purified phospholipase A2 from the venom of the Papuan black snake (Pseudechis papuanus).

A neutral phospholipase A2 (PLA2) was separated from Pseudechis papuanus venom by a two-stage FPLC procedure of cation exhange and phenyl-Superose chromatography. It had a molecular mass of 15 kDa and a lower LD50 value than a co-separated haemorrhagic fraction, indicating a higher lethal potency. In vitro tests confirmed the powerful inhibition of platelet aggregation by the PLA2 and strong anticoagulant activity initially observed with whole venom. Ultrastructural studies showed that platelets lost their discoid shape and developed membranous projections with a general decrease in electron-density of the cytosol and disruption of the microfilaments following incubation with the enzyme. Amino acid sequence analysis of the N-terminus and some internal peptides demonstrated a high degree of homology with PLA2s from other Pseudechis venoms. Our results indicate that this fraction is the main agent responsible for the haemostatic disorders in envenomed patients.

Aged Garlic Extract May Inhibit Aggregation in Human Platelets by Suppressing Calcium Mobilization

Cardiovascular disease is associated with multiple factors including the increased ability of platelets to aggregate. Aged garlic extract (AGE) was shown to inhibit platelet aggregation; however, the underlying mechanisms have yet to be established. Because calcium mobilization plays an important role in platelet aggregation, the effect of AGE was investigated in this preliminary study. ADP and the calcium ionophore A23187 both stimulated platelet aggregation with a concomitant increase in intracellular calcium ion concentration. When these experiments were repeated in the presence of AGE, both platelet aggregation and calcium mobilization were suppressed. In addition, when platelets were preincubated with AGE, the initial concentration of intracellular calcium was significantly reduced compared with platelets without AGE, confirming the metal-chelating properties of AGE. Platelets loaded with fura-2 acetoxymethyl ester (fura-2 AM) also displayed a reduction in platelet aggregation, and the addition of external calcium did not alter this observation. Although variable data were obtained in this study, these results taken together imply that AGE probably exerts its inhibitory effect on platelet aggregation either by suppressing the influx of calcium ions by chelating calcium within platelet cytosol or by altering other intracellular second messengers within the platelets.