Effect of the Magnetized Water Supplementation on Lymphocyte DNA Damage in Mice Treated with Diethylnitrosamine

Water gets magnetically charged when it is contacted with a magnet. Although magnetic water products have been promoted since the 1930's, they have received very little recognition due to questionable effectiveness. Diethylnitrosamine (DEN) is a widely occurring nitrosamine that is one of the most important environmental carcinogens primarily inducing tumors of liver. In this study, the effect of magnetized water supplementation on lymphocyte DNA damage in ICR mice treated with DEN was evaluated using the Comet assay. Mice were divided into 3 groups: control, DEN, and DEN + magnetized water group. Fifteen mice were maintained in each group for the entire experimental period of 6, 12 and 18 weeks. Five mice in each group were sacrificed at 6, 12, and 18th weeks, followed by the Comet assay using the blood obtained from heart puncture of the mice. The level of lymphocyte DNA damage reflected by tail moment and other DNA damage indices of tail DNA (%) or tail length of the magnetized water group were significantly decreased after the 6th, 12th and 18th weeks of supplementation compared with the positive control, the DEN group. The relative DNA damage of the magnetized water groups compared to the DEN control group after 6th, 12th, and 18th weeks of supplementation were 42.2%, 40.8%, and 32.9% for DNA in tail, 31.2%, 32.6%, and 21.3% for tail length, and 33.8%, 33.8%, and 24.6% for tail moment, respectively. This is the first report demonstrating that magnetized water may be involved in the lowering effect of the DNA damage in DEN-treated ICR mice. This result suggests that the magnetized water might have minimized the DNA damage by improving the antioxidant status of the mice. However, further studies are needed to characterize the condition of the magnetization and examine the long-term effect of the water product.

The Improvement of Chaga Mushroom (Inonotus Obliquus) Extract Supplementation on the Blood Glucose and Cellular DNA Damage in Streptozotocin-Induced Diabetic Rats

Mushrooms have become a largely untapped source of powerful new pharmaceutical products that poses anti-inflammatory, and antimutagenic, and antioxidant activities. The antioxidant effects of the mushroom may be partly explained by protecting cellular components against free radical. The aim of this study was to investigate the protective effect of chaga mushroom against diabetes, via the mitigation of oxidative stress and reduction of blood glucose, in streptozotocininduced diabetic rats. Rats were rendered diabetic by intravenous administration of STZ through tail at a dose of 50 mg/kg. Animals were allocated into four groups with 8 rats each. The control and diabetic control group were fed withstandard rat feed. The other diabeic groups, the low chaga extract group and the high chaga extract group were fed ad libitum using 0.5 g/kg and 5 g/kg of chaga mushroom extract, respectively, for 4 weeks. The blood glucose levels in the two chaga extract groups showed a tendency to decrease but did not reach statistical significance after the supplementation. Leukocyte DNA damage, expressed as tail length, was found to be significantly lower in the high chaga extract group than in the diabetic control group (p > 0.05). Plasma level of total radical-trapping antioxidant potential (TRAP) was tend to be higher in the high chaga extract group compared with the diabetic control group. Erythrocyte antioxidant enzyme activities of two groups did not differ. Although we did not obtain beneficial effect on lowering blood glucose levels in the STZ-induced diabetic rats, this results suggest that the chaga mushroom extracts may initially act on protecting endogenous DNA damage in the short-term experiment.

Effect of Deer Antler Drink Supplementation on Blood Pressure, Blood Glucose and Lymphocyte DNA Damage in Type 2 Diabetic Patients

Deer Antler has been known for its traditional oriental medicinal properties and has been widely used to promote growth, boost immune function, treat blood loss and chronic joint pain. Recent study showed imported (New Zealand) Deer Antler was beneficial in reducing the side effects of cancer treatments. However, there was no intervention study conducted on the effect of Korean Deer Antler on reducing the oxidative stress to patients with diabetes. One of the sensitive ways to measure endogenous oxidative stress is by measuring cellular DNA damage using single cell gel electrophoresis (COMET assay). This study was conducted to investigate the possible beneficial effect of commercial Deer Antler drink (provided by Chung-yang Deer Farm) on lymphocyte DNA damage and blood glucose of diabetic patients. Ten patients (4 men, 6 women) participated in the study and consumed 2 pouches of Deer Antler drink every day for 20 days. Blood was collected on the morning before and after the intervention for lymphocyte isolation and blood glucose analysis. Both systolic and diastolic blood pressure showed a tendency to decrease but did not reach statistical significance after the trial. Blood glucose level was not affected by the supplementation. After the intervention, over 50% reduction were noted in the cellular DNA damage, expressed as tail length (TL) and tail moment (TM; tail length x percent tail DNA). Although we did not obtain beneficial effect on lowering blood glucose levels in the patients, this results suggest that Deer Antler may initially act in protecting endogenous DNA damage in short-term experiment.

Comparison of the Protective Effect of Antioxidant Vitamins and Fruits or Vegetable Juices on DNA Damage in Human Lymphocyte Cells Using the Comet Assay

In this study the in vitro protective effects of several antioxidant vitamins (vitamin C, alpha-tocopherol, beta-carotene), fruits and vegetables (strawberry, tangerine, orange and 100% orange juice, carrot juice), on the levels of isolated human lymphocyte DNA damage was measured using Comet assay. Comet assay has been used widely to assess the level of the DNA damage in the individual cells. Lymphocytes were pre-treated for 30 minutes with antioxidant vitamins (10, 50, 100, 500 micrometer) or fruits. vegetables (10, 100, 500, 1000 microgram/ml), an4 then oxidatively challenged with 100 micrometer H2O2 for 5 min at 4degrees C. The protective effect of antioxidant vitamins against DNA damage at a concentration of 50 micrometer were 50% in vitamin C, 32% in alpha-tocopherol, whereas, beta-carotene showed a 55% protection at a dose as low as 10micrometer. The inhibitory effects of DNA damage by strawberry, tangerine, orange, orange juices, carrot juices were 50 - 60% with wide ranges of doses. The results of the present study indicate that most the antioxidant vitamins and fruits.vegetables juices produced a significant reduction in oxidative DNA damage.

The Effects of Purple Grape Juice Supplementation on Improvement of Antioxidant Status and Lymphocyte DNA Damage in Korean Smokers

The purpose of this project was to evaluate whether daily fruit juice consumption could reduce the DNA damage in healthy subjects. The study was performed using 67 healthy volunteers (29 smokers, 38 nonsmokers) who were supplemented with 480 ml of grape juice for 8 weeks. Eight weeks of grape juice consumption did not change any anthropometric parameters. Lymphocyte DNA damage before the study was significantly greater (p < 0.05) in smoker than nonsmoker, but, grape juice consumption significantly reduced DNA damage in both smoker (26%) and nonsmoker (17%) to the level where there was no difference remained between the two groups after the intervention trial. This preventive effect of grape juice against DNA damage was not affected by sex of the subjects in non-smokers. Plasma alpha-carotene, lycopene and gamma-totopherol was significantly increased after the trial in smokers, while erythrocyte catalase was significantly increased in both smokers and nonsmokers. Total radical-trapping antioxidant potential (TRAP) level in all subjects was significantly reduced after the intervention, while GSH-Px activity was increased only in nonsmokers. These results suggests that daily consumption of grape juice may protect DNA damage in peripheral lymphocytes, and supports the hypothesis that grape juice might exert their effect partially via a decrease in oxidative damage to DNA in humans partly by improving their antioxidative defense system.

Protective Effect of Yellow-Green Vegetable Juices on DNA Damage in Chinese Hamster Lung Cell Using Comet Assay

The present study was attempted to investigate the antioxidant capacity of popular yellow-green vegetable juices (kale, Angelica keishei, carrot, small water dropwort) and to investigate the effect of vegetable juices on protecting oxidative damage to DNA in cultured Chinese hamster lung (CHL) cells. Antioxidant capacity was analyzed by TRAP assay (Total radical-trapping antioxidant potential). Cellular DNA dmamage was measured by SCGE (single-cell gel electrophoresis, also known as comet assay. Cells incubated in medium with PBS (negative control) or with various concentration of the freeze dried green juices (25, 50, 100, 250 ug/mL) resuspended in PBS were treated with H2O2 (200 micrometer) as an oxidative stimulus for 5 min at 4 degrees C. The physiological function of each vegetable juice on oxidative DNA damage was analyzed and expressed as tail moment (tail length X percentage migrated DNA in tail). Kale juice had the highest TRAP value suggesting that kale has the highest antioxidant capacity followed by Angelica keishei, small water dropwort and carrot. Cells treated with H2O2 had extensive DNA damage compared with cells treated with PBS or pre-treated with vegetable juice extracts. All green juices inhibited H2O2-induced DNA damage with kale being the most effective juice among the tested juices. These results indicate that green juice supplementation to CHL cells followed by oxidative stimulus inhibited damage to cellular DNA, supporting a protective effect against oxidative damage induced by reactive oxygen species.

Protective Effect of Flavonoids on Lymphocyte DNA Damage Using Comet Assay

The present study was attempted to investigate and compare the antioxidant potency of several well-know flavonoids, antioxidant vitamin and commercially available popular beverages. The antioxidant potency was assessed by the effect on reducing oxidative DNA damage of human lymphocytes. Cellular oxidative DNA damage was measured by SCGE (single-cell gel electrophoresis), also known as comet assay. Lymphocytes were pre-treated for 30 minutes with wide ranges of doses of apigenin, kaempferol, luteolin, myricetin, rutin, quercetin, alpha-tocopherol (10,25,50,100,200,500,1000 micrometer) ,green tea extract or grape juice (10,50,100,250,500,1000 microgram/mL) followed by a H2O2(100 micrometer) treatment for 5 min as an oxidative stimulus. The physiological function of each antioxidant substance on oxidative DNA damage was analyzed as tail moment (tail length X percentage migrated DNA in tail) and expressed as relative DNA damage score after adjusting by the level of control treatment. Cells treated with H2O2 alone (positive control) had an extensive DNA damage compared with cells treated with phosphate buffered saline (PBS, negative control) or pre-treated with all the tested samples. Of all the six flavonoids, quercetin was the most potent antioxidant showing the lowest ED50 of 8.5 microgram/mL (concentration to produce 50% protection of relative DNA damage). The antoxidant potency of individual flavonoids were ranked as follows in a decreasing order; luteolin (18.4 microgram/mL), myricetin (19.0 microgram/mL) , rutin (22.2 microgram/mL) , apigenin (24,3 microgram/mL) , kaempferol (25.5 microgram/mL). The protective effect of alpha-tocopherol was substantially lower (highest ED50 value of 55.0 microgram/mL) than all the other flavonoids, while the protective effect was highest in green tea and grape juice with low ED5O value of 7.6 and 5.3, respectively. These results suggest that flavonoids, especially quercetin, and natural compounds from food product, green tea and grape juice, produced powerful anti-oxidative activities, even stronger than alpha-tocopherol. Taken together, supplementation of antioxidants to lymphocytes followed by oxidative stimulus inhibited damage to cellular DNA, supporting a protective effect against oxidative damage induced by reactive oxygen species.