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Direct inhibition of MTX on NADPH (NADPH is a cofactor for enzyme GSH reductase to make reduced GSH) together with overproduction of ROS, will cause a reduction in GSH cellular availability.[ , , , ] Furthermore, the influence of MTX on Nrf2 expression might also reduce GSH tissue level.[] SOD is an essential endogenous antioxidant enzyme, induces the conversion of mitochondrial generated ROS, mainly superoxide (O2), into less toxic hydrogen peroxide (H2O2) or molecular oxygen (O2).[] On the other hand, MTX causes mitochondrial membrane damage, and more superoxide (O2) free radical production will lower SOD activity.[] Overproduction of ROS and loss of endogenous antioxidants will shift the oxidative balance toward oxidative stress