C-phycocyanin及其亚基对H2O2诱导GC-1 spg细胞程序性坏死的保护作用

Abstract

C-phycocyanin (C-PC) is an effective antioxidant and has an important value in medical research. Oxidative stress is considered to be one of the main underlying mechanisms of cell death, and reducing oxidative stress is one of the strategies to enhance germ cell viability. Herein, we investigated the protective effect and the mechanism of C-PC and apo-phycocyanin subunit on oxidative stress damage induced by H 2 O 2 in GC-1 spg cells. C-PC genes were cloned into the pGEX-4T-1 vectorand transformed into Escherichia coli BL21 to achieve the efficient expression of C-PC subunit. GC-1 spg cells were treated with 600 μM H 2 O 2 for 24 h to establish the oxidative stress damage model. Cell viability was detected by CCK-8. The degree of oxidative stress was detected by testing Superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities and glutathione (GSH) and Malondialdehyde (MDA) levels. Reactive oxygen species (ROS) was evaluated utilizingby 2', 7'-dichlorofluorescent-diacetate (DCFH-DA). Mitochondrial membrane potential was determined by JC-1. Cell necrosis rate was detected by Annexin V-FITC/PI. Expression of protein was detected by western blot. We found that C-PC and GST-CPC β significantly inhibited H 2 O 2 -induced oxidative damage of GC-1 spg cells, improved the ability of antioxidation, reduced ROS overproduction, and mitochondrial membrane potential loss, and inhibited the RIP-1/RIP-3/ p-MLKL signaling pathway to reduce the necrosis rate. The results demonstrated that C-PC played a protective role against H 2 O 2 -induced cell damage, especially its β subunit. This study provides a theoretical basis for C-PC as a potential protective agent of reproductive system.