Hydroxytyrosol, the phenolic compound of olive oil protects human PBMC against oxidative stress and DNA damage mediated by 2,3,7,8-TCDD

The protective effect of hydroxytyrosol (HT), a strong antioxidant compound from extra virgin olive oil, against TCDD induced toxicity was investigated in human peripheral blood mononuclear cells (PBMC). PBMC (1 × 10⁶ cells mL⁻¹) were divided into four groups and were incubated in a CO₂ incubator (5% CO₂) for 12 h with vehicle, TCDD (10 nM), TCDD + HT (10 nM + 100 μM) and HT alone (100 μM) respectively. To clarify the role of HT against TCDD induced cytotoxicity, oxidative stress and the levels of antioxidant enzymes were assessed. Incubation of PBMC with TCDD significantly decreased cell viability, catalase (CAT) and glutathione peroxidase (GPx) and increased the levels of superoxide dismutase (SOD), glutathione reductase (GR) and oxidative stress markers such as lipid peroxidation products (LPO), protein carbonyl content (PCC) and reactive oxygen species (ROS). Whereas, HT had an effective antioxidant property as observed by the increased cell viability, normalization of antioxidant enzymes and decreased levels of LPO, PCC and ROS in PBMC co-treated with HT and TCDD. Apoptosis detection and comet assay results shows that HT, by acting as an antioxidant, prevents the damage to DNA induced by TCDD. In addition light microscopic and histopathological observations revealed that the cells are apoptotic and degenerated during TCDD treatment, whereas cells showed intact morphology during co-treatment with HT. On the whole, the results reveal that HT exerts a promising antioxidant potential in protecting the PBMC against TCDD induced oxidative stress, which might be due to the presence of catechol moiety in its structure.     

Protective effect of silymarin on erythrocyte haemolysate against benzo(a)pyrene and exogenous reactive oxygen species (H2O2) induced oxidative stress

The present study was carried out to evaluate the in vitro antioxidant properties and protective effects of silymarin (milk thistle) in human erythrocyte haemolysates against benzo(a)pyrene [B(a)P], a potent carcinogenic chemical. Protective effect of silymarin was assessed in vitro by monitoring the antioxidant enzymes and malondialdehyde in three groups of haemolysates-(I) vehicle control (II) B(a)P incubated group and (III) B(a)P co incubated with silymarin. The effects of silymarin on lipid peroxidation (LPO) and antioxidant enzymes [superoxide dismutase; SOD, catalase; CAT, glutathione peroxidase; GPx, glutathione reductase; GR and glutathione-S-transferases; GST] were assessed on haemolysates. It was observed that specific activity of antioxidant enzymes were significantly decreased and the malondialdehyde levels were elevated when haemolysates were incubated with B(a)P. The protective effect of silymarin is elucidated by the significant reversal of the antioxidant enzymes and reduction in the levels of malondialdehyde. In addition, haemolysates were incubated with B(a)P for 45 min and the B(a)P metabolite, 3-hydroxy benzo(a)pyrene (3-OH-B(a)P) was detected using HPLC. An increased level of the metabolite was detected in group II. Whereas, when haemolysates were co-incubated with silymarin, the reactive metabolite 3-OH-B(a)P was not detectable which further confirms the protective role of silymarin. Generation of 3-OH-B(a)P in group II implicates the possibility of reactive oxygen species (O2- and H2O2) production in haemolysates during cytochrome P4501A1 (CYP1A1) mediated Phase-I-metabolism. Hence, we incubated the haemolysates with exogenous reactive oxygen species H2O2 and assessed the protective role of silymarin against H2O2. From the results of our study, it was suggested that silymarin possess substantial protective effect and free radical scavenging mechanism against environmental contaminants induced oxidative stress damages.