Comparison of catechin profiles in human plasma and urine after single dosing and regular intake of green tea (Camellia sinensis).

Green tea (Camellia sinensis) catechin profiles in plasma and urine following single dosing and regular ingestion of green tea are not clear. We performed a placebo-controlled intervention study with sixteen healthy volunteers to determine changes in total and free catechins after a single dose and following 1 week of twice-daily green tea. Blood and urine samples were collected before (fasting) and after (60 and 120 min for blood; 90 and 180 min for urine) drinking 200 ml of 1.5% (w/v) green tea or water (n 8 each), and fasting samples were again collected after 7 d of 150 ml of 1% (w/v) supplemental green tea or water twice daily. After a 4-week washout, subjects were crossed onto the other treatment and procedures repeated. Plasma results at 1 h post-ingestion showed elevated (P < 0.05) mean epigallocatechin gallate (EGCG; 310 (SD 117) nmol/l; all in free form), epigallocatechin (EGC; 192 (SD 67) nmol/l; 30% free) and epicatechin gallate (ECG; 134 (SD 51) nmol/l; 75% free). Fasting plasma after 7 d of regular intake showed increased (P < 0.05) EGCG (80 v. 15 nmol/l at baseline) and ECG (120 v. 40 nmol/l), with > or =90% of both in their conjugated forms. Total EGC was < 10 nmol/l. Post-ingestion conjugation and renal loss of EGC and epicatechin were rapid and high, but were negligible for EGCG and ECG. In the green tea consumed, the content was EGCG > EGC > ECG, and the acute plasma response mirrored this. However, after chronic consumption there was almost no EGC found in fasting plasma, some EGCG was present, but a rather high level of ECG was maintained.

Antioxidant properties of drugs used in Type 2 diabetes management: could they contribute to, confound or conceal effects of antioxidant therapy?

OBJECTIVES: This is a narrative review, investigating the antioxidant properties of drugs used in the management of diabetes, and discusses whether these antioxidant effects contribute to, confound, or conceal the effects of antioxidant therapy. METHODS: A systematic search for articles reporting trials, or observational studies on the antioxidant effect of drugs used in the treatment of diabetes in humans or animals was performed using Web of Science, PubMed, and Ovid. Data were extracted, including data on a number of subjects, type of treatment (and duration) received, and primary and secondary outcomes. The primary outcomes were reporting on changes in biomarkers of antioxidants concentrations and secondary outcomes were reporting on changes in biomarkers of oxidative stress. RESULTS: Diabetes Mellitus is a disease characterized by increased oxidative stress. It is often accompanied by a spectrum of other metabolic disturbances, including elevated plasma lipids, elevated uric acid, hypertension, endothelial dysfunction, and central obesity. This review shows evidence that some of the drugs in diabetes management have both in vivo and in vitro antioxidant properties through mechanisms such as scavenging free radicals and upregulating antioxidant gene expression. CONCLUSION: Pharmaceutical agents used in the treatment of type 2 diabetes has been shown to exert an antioxidant effect..

Effects of single dose and regular intake of green tea (Camellia sinensis) on DNA damage, DNA repair, and heme oxygenase-1 expression in a randomized controlled human supplementation study.

Regular intake of green tea (Camellia sinensis) lowers DNA damage in humans, but molecular mechanisms of genoprotection are not clear. Protection could be via direct antioxidant effects of tea catechins, but, paradoxically, catechins have pro-oxidant activity in vitro, and it is hypothesized that mechanisms relate to redox-sensitive cytoprotective adaptations. We investigated this hypothesis, focusing particularly on effects on the DNA repair enzyme human oxoguanine glycosylase 1 (hOGG1), and heme oxygenase-1, a protein that has antioxidant and anti-inflammatory effects. A randomized, placebo-controlled, human supplementation study of crossover design was performed. Subjects (n = 16) took a single dose (200 mL of 1.5%, w/v) and 7-days of (2 × 200 mL 1%, w/v per day) green tea (with water as control treatment). Lymphocytic DNA damage was ∼30% (p < 0.001) lower at 60 and 120 min after the single dose and in fasting samples collected after 7-day tea supplementation. Lymphocytic hOGG1 activity was higher (p < 0.0001) at 60 and 120 min after tea ingestion. Significant increases (p < 0.0005) were seen in hOGG1 activity and heme oxygenase-1 after 7 days. Results indicate that molecular triggering of redox-sensitive cytoprotective adaptations and posttranslational changes affecting hOGG1 occur in vivo in response to both a single dose and regular intake of green tea, and contribute to the observed genoprotective effects of green tea.

Genoprotection and genotoxicity of green tea (Camellia sinensis): Are they two sides of the same redox coin?

OBJECTIVES: Regular intake of green tea associates with lower DNA damage and increased resistance of DNA to oxidant challenge. However, in vitro pro-oxidant effects of green tea have been reported. Both effects could be mediated by hydrogen peroxide (H2O2) which is generated by autoxidation of tea catechins. In large amounts, H2O2 is genotoxic, but low concentrations could activate the redox-sensitive antioxidant response element (ARE) via the Keap-1/Nrf2 redox switch, inducing genoprotective adaptations. Our objective was to test this hypothesis. METHODS: Peripheral lymphocytes from healthy volunteers were incubated for 30 minutes at 37°C in freshly prepared tea solutions (0.005, 0.01, 0.05%w/v (7, 14, 71 µmol/l total catechins) in phosphate buffered saline (PBS), with PBS as control) in the presence and absence of catalase (CAT). H2O2 in tea was measured colorimetrically. Oxidation-induced DNA lesions were measured by the Fpg-assisted comet assay. RESULTS: H2O2 concentrations in 0.005, 0.01, and 0.05% green tea after 30 minutes at 37°C were, respectively, ∼3, ∼7, and ∼52 µmol/l. Cells incubated in 0.005 and 0.01% tea showed less (P < 0.001) DNA damage compared to control cells. Cells treated with 0.05% green tea showed ∼50% (P < 0.001) more DNA damage. The presence of CAT prevented this damage, but did not remove the genoprotective effects of low-dose tea. No significant changes in expression of ARE-associated genes (HMOX1, NRF2, KEAP1, BACH1, and hOGG1) were seen in cells treated with tea or tea + CAT. CONCLUSION: Genoprotection by low-dose green tea could be due to direct antioxidant protection by green tea polyphenols, or to H2O2-independent signalling pathways.

Cryopreservation and storage effects on cell numbers and DNA damage in human lymphocytes.

The comet assay measures DNA damage in individual cells (usually lymphocytes) and is widely used in biomonitoring studies. Lymphocytes are harvested and are usually cryopreserved for batch testing. We investigated cell loss during harvesting, cryopreservation, thawing, and washing of human peripheral lymphocytes and compared DNA damage, using the Fpg-assisted comet assay for oxidation-induced DNA lesions, in freshly harvested cells and cells that were thawed and tested after cryopreservation of 2-3 days and 4 weeks. Lymphocyte numbers were measured in fresh venous blood and after the steps of harvesting, cryopreservation, and washing. Results showed that >50% of lymphocytes in whole blood were harvested, but ∼60% were lost during washing. Loss during washing was not different (P>0.05) between fresh cells and cells thawed and washed after 2-3 days or 4 weeks cryopreservation. No change in DNA damage was seen after cryopreservation and thawing: mean (SD) % DNA in comet tail was 11.2 (1.53) in freshly harvested cells, 12.9 (1.39) in 2-3 days cryopreserved cells, and 12.9 (2.0) in cells tested after 4 weeks cryopreservation (P>0.05). Results indicate that there is no predominant loss of more highly damaged cells during cryopreservation and thawing and there is no induction of oxidation-induced DNA lesions in cryopreserved cells stored for up to 4 weeks.