Chemopreventive Effects of β‐Carotene, α‐Tocopherol and Five Naturally Occurring Antioxidants on Initiation of Hepatocarcinogenesis by 2‐Amino‐3‐methylimidazo[4,5‐f] qumoline in the Rat

Inhibitory effects of naturally occurring antioxidants on the initiation stage of hepatocarcinogenesis were studied. Group 1 rats were given a diet containing β‐carotene (β8‐CT, 0.02%), α‐tocopherol (α‐TP, 1.5%), glutathione (GLT, 5%), vanillin (VNL, 1%), quercetin (QCT, 1%) or ellagic acid (ELA, 1%), or 3 doses of diallyl sulfide (DAS, 200 mg/kg, i.g) over an 8‐day period. On day 7, the animals received a single dose of 2‐amino‐3‐methylimidazo[4,5‐f]jumoline (IQ, 100 mg/kg, i.g.), 12 h after two‐thirds partial hepatectomy for initiation and 2 weeks thereafter, were placed on promotion regimen comprising phenobarbital (0.05% in diet) and a single dose of D‐galactosamine (100 mg/kg, i.p.). Groups 2 and 3 were treated as described for Group 1, but without test material or IQ, respectively. Survivors were killed at week 11 and antioxidant influence was assessed by comparing values for preneoplastic glutathione S‐transferase placental form‐positive (GST‐P(+)) foci between Groups 1 and 2. All lesions larger than 70 βm in diameter consisting of approximately 5 cells in cross section were counted. Numbers of GST‐P+ foci/cm(2) in Group 1 were: β ‐CT, 7.99; α‐TP, 8.21; GLT, 9.71; DAS, 10.37; VNL, 10.57; QCT, 11.1; ELA, 12.5 (n = 11‐15). All, except ELA, showed a significant decrease as compared with the Group 2 value of 14.54 (n=15). Only β ‐CT showed a significant decrease for the area value. This is the first report to show that β ‐CT, α‐TP, GLT, DAS, VNL, QCT exert inhibitory effects on initiation of hepatocarcinogenesis by the food carcinogen IQ, suggesting that these antioxidants might find application as chemopreventive agents. Furthermore, the current protocol proved practical for the assessment of chemopreventive agents within 11 weeks, a relatively Short period.