Genotoxic and antigenotoxic effects of catechin and tannins from the bark of Hamamelis virginiana L. in metabolically competent, human hepatoma cells (Hep G2) using single cell gel electrophoresis.
The genotoxic and antigenotoxic activities of catechin, hamamelitannin and two proanthocyanidin fractions prepared from the bark of Hamamelis virginiana L. were investigated in a human derived, metabolically competent hepatoma cell line (Hep G2) using single cell gel electrophoresis (SCGE) for the detection of DNA-damage. DNA-migration was calculated as Olive tail moment (OTM). Catechin and a low-molecular weight proanthocyandin fraction (W(M)) caused only slight increases of OTM up to concentrations of 166 microg/ml whereas hamamelitannin and the proanthocyandin fraction with higher molecular weight (W(A)) led to a two-fold enhancement of OTM at the same concentrations. These effects were dose-independent. Treatment of the cells with the test compounds in a dose-range of 2-166 microg/ml prior to the exposure to benzo(a)pyrene (B(a)P, 10 microM, 2.5 microg/ml) led to a significant reduction of induced DNA damage which was dose-dependent for all test compounds, except for hamamelitannin. The inhibitory effects of proanthocyanidins were stronger than those of catechin and hamamelitannin; the lowest effective concentrations were about 2 microg/ml. In order to clarify the mechanisms of protection, possible effects of the test compounds on enzymes involved in toxification and detoxification of B(a)P were investigated. While B(a)P toxification by cytochrome P450 was not inhibited by the test compounds, detoxification by glutathion-S-transferase (GST) was induced by catechin and W(M). Combination experiments with the ultimate metabolite of B(a)P, (+/-)-anti-benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide (BPDE; 5 microM, 1.5 microg/ml), revealed strong inhibitory effects, indicating that the observed protective effects were caused by scavenging of the ultimate mutagen by the test compounds. Exposure of Hep G2 cells to the test compounds after B(a)P treatment did not influence B(a)P induced DNA damage, demonstrating that repair mechanisms were not affected.
Dauer
Andreas
A
Hochschule Wädenswil, University of Applied Sciences, Pharmaceutical Biotechnology, Box 335, Gruental, CH-8820 Wädenswil, Switzerland.
Hensel
Andreas
A
Lhoste
Evelyne
E
Knasmüller
Siegfried
S
Mersch-Sundermann
Volker
V
D000872:Anthocyanins / Q000302:isolation & purification / Q000494:pharmacology / Q000633:toxicity; D000972:Antineoplastic Agents, Phytogenic / Q000302:isolation & purification / Q000494:pharmacology / Q000633:toxicity; D001564:Benzo(a)pyrene / Q000378:metabolism / Q000633:toxicity; D006528:Carcinoma, Hepatocellular / Q000235:genetics; D002392:Catechin / Q000302:isolation & purification / Q000494:pharmacology* / Q000633:toxicity; D019363:Cytochrome P-450 CYP1A1 / Q000378:metabolism; D004249:DNA Damage; D004587:Electrophoresis, Agar Gel / Q000379:methods; D005707:Gallic Acid / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000494:pharmacology* / Q000633:toxicity; D029501:Hamamelis / Q000737:chemistry*; D006601:Hexoses / Q000302:isolation & purification / Q000494:pharmacology* / Q000633:toxicity; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics; D009153:Mutagens / Q000302:isolation & purification / Q000494:pharmacology* / Q000633:toxicity; D024301:Plant Bark / Q000737:chemistry; D044945:Proanthocyanidins; D014407:Tumor Cells, Cultured
D000872:Anthocyanins; D000972:Antineoplastic Agents, Phytogenic; D006601:Hexoses; D009153:Mutagens; D044945:Proanthocyanidins; C013221:proanthocyanidin; D001564:Benzo(a)pyrene; D005707:Gallic Acid; D002392:Catechin; D019363:Cytochrome P-450 CYP1A1; C093412:hamamelitannin