Investigation Effect of N, N′-Dipyridoxyl (Ophenylenediamine) Schiff Base on GlutathionePeroxidase Gene Expression in HepG2 Gells

Schiff base with donors (N,O,S) have structural similarities with natural biological systemsand imports in elucidating the mechanism of transformations and reseminations reactions inbiological systems due to presence of imine (-N=CH-) group. Schiff base are known to havebiological activities such as antibacterial, antifungal, antitumor and antioxidant activities. Thisstudy evaluated the role schiff base on the expression of glutathione peroxidase antioxidantenzyme. RNA was isolated from HepG2 cells (treated and untreated) using high pure RNAisolation kit (Roche, Germany) agreeing to the company’s instruction and this RNA wasused to synthesize of Complementary DNA. CDNA was synthesized using Oligo (dT),Primer, 5X Reaction Buffer, RiboLock RNase Inhibitor, dNTP, RevertAid RT, water nucleasefreeand incubation for 60 min at 42°C and 70°C for 5 min in Biotech Thermal Cycler(London, England). To perform the Real time PCR used Bio-Rad CFX96 detection system.Reaction solution for Real time PCR was containing CDNA, SYBR Green Master Mix,primer, and water. In this way, evaluated changes in level expression of glutathioneperoxidase (GPx) gene in treated cells with the different concentrations of Schiff base (250,500, 1000 Âμg/ml) as compared to untreated cells. GAPDH was used as housekeepinggene. The Real time PCR result showed that the level expression of GPx in exposed cellscompared to untreated cells considerably increased to 2.6, 5.1, 6.3 fold in 250, 500 and 1000μg/ml concentrations of schiff base. Results show that N, N′-dipyridoxyl(Ophenylenediamineschiff base can be considered as an antioxidant agent.

کلیدواژه ها:

Schiff Base ، Real Time PCR ، Glutathione Peroxidase ، Expression ، HepG2 Cells