Effects of various plant polyphenols on bladder carcinogen benzidine-induced mutagenicity

Benzidine (Bz), a human bladder carcinogen, was strongly mutagenic to Salmonella TA102 tester strain in the Ames Salmonella microsome/mutagenicity assay in the presence of rat liver S9 mix. Various non-mutagenic plant polyphenols were included in the assay to test their inhibitory effects on the Bz-induced mutations. Coumestrol, ellagic acid (EA), (-)-epicatechin (EC), (-)-epichatechingallate (ECG), gallic acid (GA), (-)-gallocatechin (GC), plumbagin, propyl gallate (PG), taxifolin, and 2,2',4'-trihydroxychalcone were found to have a strong inhibitory effect on Bz-induced mutations. (-)-Epigallo-catechingallate (EGCG), fisetin, (-)-gallocatechingallate (GCG), and piceatannol were moderately inhibitory to the mutations; whereas, (-)-catechin, (-)-catechingallate (CG), and reseveratrol were weakly inhibitory to the mutations. (-)-Epigallocatechin (EGC) and 7,3',4'-trihydroxy isoflavon were not inhibitory to the Bz-induced mutations. Isoliquirtigenin, quercetin dihydrate, and rhein were found to be mutagenic in tester strain TA102. Benzidine mediated lipid peroxidation was conducted employing the thiobarbituric acid reactive substances (TBARS) assay using linoleic acid as a substrate. In the presence of rat liver S9 mix, Bz could cause lipid peroxidation as an outcome of production of oxygen free radicals. Incorporation of the above mentioned non-mutagenic plant polyphenols significantly inhibited benzidine mediated lipid peroxidation in a time dependent manner. These polyphenols also effectively reduced the iron mediated lipid peroxidation. Thus, it is concluded that the inhibition of oxidative mutagenicity of Bz by plant polyphenols could be due to an inhibitory effect of plant polyphenols on the bioactivating enzymes such as cytochrome P-450 and peroxidase and the chelation of iron present in the cytochrome P-450 in the S9 mix. Thus, these plant polyphenols play a significant inhibitory role on Bz-induced mutagenicity.