Proton-Induced X-ray Emission (PIXE) Analysis and DNA-chain Break study in rat hepatocarcinogenesis: A possible chemopreventive role by combined supplementation of vanadium and beta-carotene.

Combined effect of vanadium and beta-carotene on rat liver DNA-chain break and Proton induced X-ray emission (PIXE) analysis was studied during a necrogenic dose (200 mg/kg of body weight) of Diethyl Nitrosamine (DENA) induced rat liver carcinogenesis. Morphological and histopathological changes were observed as an end point biomarker. Supplementation of vanadium (0.5 ppm ad libitum) in drinking water and beta-carotene in the basal diet (120 mg/Kg of body weight) were performed four weeks before DENA treatment and continued till the end of the experiment (16 weeks). PIXE analysis revealed the restoration of near normal value of zinc, copper, and iron, which were substantially altered when compared to carcinogen treated groups. Supplementation of both vanadium and beta-carotene four weeks before DENA injection was found to offer significant (64.73%, P < 0.001) protection against generation of single-strand breaks when compared with the carcinogen control counter parts. A significant stabilization of hepatic architecture of the cells was observed as compared to carcinogen control in vanadium plus beta-carotene treated group. This study thus suggests that vanadium, a prooxidant but potential therapeutic agent yield safe and effective pharmacological formulation with beta-carotene, an antioxidant, in the inhibition of experimental rat hepatocarcinogenesis.

Combined supplementation of vanadium and beta-carotene suppresses placental glutathione S-transferase-positive foci and enhances antioxidant functions during the inhibition of diethylnitrosamine-induced rat liver carcinogenesis.

BACKGROUND AND AIM: The present study was designed to investigate the chemopreventive effects of combined vanadium (V; 0.5 p.p.m.) and beta-carotene (BC; 120 mg/kg of basal diet) on diethylnitrosoamine (DEN)-induced and phenobarbital (PB)-promoted rat hepatocarcinogenesis. METHODS: All rats were subjected to two-thirds partial hepatectomy (PH) at the fourth week. After PH they were administered either trioctanoin alone (groups A', B', C' and D') or a single injection of DEN in trioctanoin at a dose of 10 mg/kg of body weight (groups A, B, C and D). Two weeks after the DEN treatment PB was administered (0.05% in basal diet) to all the DEN-treated rats and continued until the end of the experiment. Supplementation of V (groups B and B'), BC (groups C and C') or both V and BC (groups D and D') at the doses stated previously were started 4 weeks before DEN administration (at week 0) and continued until the 16th week. RESULTS: It was observed that in the DEN-treated and PB-promoted group (group A) the expression of the numbers and areas of the placental form of glutathione S-transferase (GST-P)-positive altered hepatic foci (AHF) was maximum. Treatment with V (group B) and BC (group C) significantly reduced the expression of GST-P-positive AHF by 29.5% and 42.8%, respectively. An additive protection action (65.7%) was noticed in group D, which received both V and BC for the entire period of the experiment. It was also observed that supplementation of V and BC for the entire period of the experiment significantly reduced the number and size of the hyperplastic nodules, while the combination treatment worked as an additive effect, reducing the number and size of the hyperplastic nodules to 22% from 89%. Moreover, a significantly reduced level of cytosolic glutathione (P < 0.001) and glutathione-S-transferase (P < 0.001) activity and stabilization of aerobic metabolism and hepatic architecture of the cells as compared with carcinogen control were observed in the V + BC-treated group. CONCLUSION: The present study suggests that V, an essential trace element, may be useful in combination with BC, an antioxidant, in the inhibition of experimentally induced rat hepatocarcinogenesis.

1alpha, 25-dihydroxyvitamin D3 prevents DNA damage and restores antioxidant enzymes in rat hepatocarcinogenesis induced by diethylnitrosamine and promoted by phenobarbital.

AIM: To investigate the chemopreventive effects of 1alpha, 25-dihydroxyvitamin D(3) in diethylnitrosamine induced, phenobarbital promoted rat hepatocarcinogenesis. METHODS: The rats were randomly divided into 6 different groups (A-F). Groups A, C and E rats received a single intraperitoneal (i.p.) injection of diethylnitrosamine (DEN) at a dose of 200 mg/kg body mass in 9 g/L NaCl solution at 4 wk of age, while group B served as normal vehicle control received normal saline once. After a brief recovery of 2 wk, all the DEN treated rats were given phenobarbital (PB) at 0.5 g/L daily in the basal diet till wk 20. Group A was DEN control. Treatment of 1alpha, 25-(OH)2D3 in group C was started 4 wk prior to DEN injection and continued thereafter till wk 20 at a dose of 0.3 microg/100 microL propylene glycol per one single dose (os) twice a week. Group E received the treatment of 1alpha, 25-(OH)2D3 at the same dose mentioned as above for 15 wk. The rats in group D and F received 1alpha, 25-(OH)2D3 alone as in group C without DEN injection. RESULTS: The comet assay showed statistically higher mean values for length to width ratios (L: W) of DNA mass and tailed cells (P<0.01; P<0.01 respectively) in DEN treated rats as compared to their normal controls. Continuous supplementation of 1alpha, 25-dihydroxyvitaminD2 showed a significant (P<0.01) decrease in L:W ratio of DNA mass tailed cells. Furthermore, 1alpha, 25-(OH)2D3 supplementations elevated the super oxide dismutase (SOD) activity, hepatic malondialdehyde (MDA) level, reduced glutathione (GSH) and glutathione S-transferase (GST) activity (P<0.01, P<0.05, P<0.05 and P<0.05 respectively). As an endpoint marker histological changes were observed to establish the chemopreventive effects of 1alpha, 25-dihydroxyvitaminD2. CONCLUSION: Supplementations of 1alpha, 25-(OH)2D3 has a marked protection against hepatic nodulogenesis, antioxidant enzymes and DNA damages in DEN induced rat hepatocarcinogenesis promoted by phenobarbital.