Is pycnogenol a double-edged sword? Cataractogenic in vitro, but reduces cataract risk in diabetic rats.

PURPOSE: Pycnogenol was used (a) to study its antioxidant activity, (b) to study its effects on lens integrity in organ culture and (c) in vivo to determine whether it could reduce the damage in model diabetic cataract. METHODS: For (a) our luminescent antioxidant assay was used, (b) lenses were incubated in medium 199, with 55.6 mM glucose. Lenses were stained with 0.014 mM rhodamine 123 for 15 min to stain mitochondria, immobilized in 1% agarose in M199, and the equatorial region examined by a Zeiss confocal microscope. For (c) cataract grades of streptozotocin diabetic rats fed 1% pycnogenol were followed for 12 weeks. RESULTS: (a) Pycnogenol in vitro was an antioxidant when challenged with peroxide. (b) In vitro, when [570 mg/L] pycnogenol in dimethyl sulfoxide (DMSO) was used, lenses turned opaque after 3 d of incubation, in both pycnogenol controls and glucose + pycnogenol. Normal controls (DMSO, n = 4) and controls (n = 4) remained clear after 8 d of incubation. After 3 d of incubation with pycnogenol, cumulative protein leakage was greater than 0.28 mg/mL versus 8 d controls (0.018 mg/mL). Similar damage occurred at pycnogenol concentrations as low as 20 mg/L. The 20 mg/L pycnogenol control showed mitochondrial death, and calcium concentration in the lens equatorial differentiating fiber cells increased. (c) In vivo feeding pycnogenol resulted in similar growth and body condition for diabetic rats, and lower cataract grades at 9 and 11 weeks: final serum glucose levels were not significantly different, but glycohemoglobin A1 levels were significantly lower (83.9% of normal, p < 0.05) in pycnogenol-fed diabetic rats. CONCLUSIONS: Although it appears that pycnogenol has a potential toxic effect on incubated lenses, it appears in vivo to have a marginal protective effect, and also significantly reduces glycation of proteins. Supported by Cognis US (formerly Henkel Chemical Co.) and Horphag Research.

Focal length variability and protein leakage as tools for measuring photooxidative damage to the lens.

Hypericin is the ingredient used to standardize the popular over-the-counter antidepressant medication St. John's Wort. Because hypericin readily produces singlet oxygen and other excited state intermediates, it is a very efficient phototoxic agent in the eye that can potentially induce the development of the cataract photooxidative mechanism. Hypericin absorbs in the UV and visible ranges, binds to the lens crystallins (alpha, beta and gamma) and damages these proteins through a photooxidative mechanism. Effects were measured previously using fluorescence, UV and mass spectrometry. We report here two additional methods to monitor lens damage: (1) measuring focal length variability using a ScanTox instrument and (2) measuring protein leakage from the damaged lens. Because nonenzymic glycation results in free radical production, we chose to use elevated glucose concentrations as a convenient model for studying oxidative stress. To compare and contrast photooxidative damage against oxidative damage to the lens, we also measured the focal length variability and protein leakage induced by the presence of elevated glucose concentrations. We found that the total accumulated protein leakage was positively correlated (r = 0.9) with variability in focal length. Lenses treated with hypericin and irradiated with UVB had an increase in focal length variability as compared with the lenses that were only UVB-irradiated. Lenses without UVB irradiation had much lower focal length variability than irradiated lenses. For non-hypericin-treated lenses, UVB-irradiated lenses had a larger variability (4.58 mm) than the unirradiated lenses (1.78 mm). The lenses incubated in elevated glucose concentrations had a focal length variability (3.23 mm) equivalent to that of the unirradiated hypericin-treated lenses (3.54 mm). We conclude that photooxidative damage by hypericin results in changes in the optical properties of the lens, protein leakage and finally cataract formation. In contrast to this, high concentrations of glucose induced protein leakage but not changes in optical properties or the opacity associated with a cataract. This work provides further evidence that people should protect their eyes from intense sunlight when taking St. John's Wort.

Phenol antioxidant quantity and quality in foods: beers and the effect of two types of beer on an animal model of atherosclerosis.

The free phenols have been measured in 15 lagers, 6 porters and ales, and 11 light and nonalcoholic beers. Phenols were measured colorimetrically using an oxidation-reduction reaction with Folin-Ciocalteu reagent and catechin as the standard. The order of phenol concentration was ales > lagers > low calorie > nonalcoholic. The quality of antioxidants of the major phenols in beers and the quality of beer antioxidants were measured by (1) dose-response inhibition of lower density lipoprotein oxidation and (2) concentration of phenols in the beers at which 50% of the peroxide was destroyed in a luminescent assay for antioxidant activity. The beers' lipoprotein antioxidant quality was clearly superior to that of vitamin antioxidants and to that of the phenol ingredients, suggesting synergism among the antioxidants in the mixture. The average per capita consumption of beer in the United States in 2000 was 225 mL/day, equivalent to 42 mg/day of catechin equivalents. Beer provides more antioxidants per day than wine in the U.S. diet. A dark beer and a lager beer were given at two concentrations to cholesterol-fed hamsters, an animal model of atherosclerosis. At the high dose ((1)/(2)-diluted beer) both lager and dark beer significantly inhibited atherosclerosis compared to a control of 2% alcohol. At the high dose, lager significantly decreased cholesterol and triglycerides, and both beers acted as in vivo antioxidants by decreasing the oxidizability of lower density lipoproteins. At the low dose ((1)/(10)-diluted beer) only the lager beer significantly decreased atherosclerosis compared to the 0.4% alcohol control. The polyphenols in the beers appear to be responsible for the benefits of beer in this model. Lager beer inhibited atherosclerosis at a human equivalent dose in this hamster model of atherosclerosis.