Evaluation of antioxidant properties of n,n n′-dipyridoxyl -(o-phenylenediamine) schiff base and its effect catalase gene expression

In modern societies exposure to contaminated environments, radiation, unhealthy foods, result in formation of free radicals which cause various diseases, including cancer. Antioxidants are group ofcompounds which scavenge free radicals and return damages tissues and cells to healthy state. Schiff base is known to have biological activities such as antibacterial, antifungal, antitumor and antioxidantactivities. the goal of this study is to evaluate antioxidant properties of Schiff base and also investigate its effect on the expression of Catalase as antioxidant enzyme. ABTS ( 2,2 -azino-bis(3- ethylbenzothiazoline-6-sulphonic acid)) solved in potassium per sulfate in order to form free radicals. This solution has the peak absorbance at 734 nm, which has green color. Antioxidant materials result indecreasing optical absorbance. In addition, HepG2 cells were treated and the RNA was isolated using high pure RNA isolation kit. Complementary DNA (cDNA) was synthesized using Oligo (dT), Primer,5X Reaction Buffer, RiboLock RNase Inhibitor, dNTP, RevertAid RT, water nuclease-free and incubation for 60 min at 42°C and 70°C for 5 min in Biotech Thermal Cycler (London, England). Then, Real time PCR was performed with reaction solution for containing CDNA, SYBR Green Master Mix, primer, and water. In this way, evaluated changes in level expression of Catalase (CAT) gene in treated cells with the different concentrations of Schiff base (250, 500, 1000 μg/ml) compared to untreated cells. GAPDH was used as housekeeping gene.The antioxidant effects of Schiff base in scavenging ABTS free radicals show that the IC50 values are 120 μg/ml.The Real time PCR result showed that the level expression of CAT in exposed cells compared to untreated cells considerably increased to 3.2, 3.01, 6 fold in 250, 500 and 1000 μg/ml concentrations of Schiff base. Conclusion: Antioxidant effects of of N,N′-dipyridoxyl (O-phenylenediamine) schiff base along with increasing the expression level of Catalase enzyme indicated this schiff base has antioxidant properties which further studies should be done in order to its usage in clinic. These results show that N,N′-dipyridoxyl (O- phenylenediamine) Schiff base can be considered as an antioxidant agent

Keywords:

Gene and Cancer , Cell and Cancer , Lifestyle and Cancer , Drugs and Cancer , Chemotherapy