Lampreys Phb2 Enhance Cellular Oxidative Stress Tolerance And Mechanism Exploration

Lampreys are unique fish animals in the northeast of China, it’s also a bridge which connecting invertebrates and vertebrates.PHB is widely distributed in a variety of biological cells and mainly anchored in the inner mitochondrial membrane as the form of heterodimers, dynamically maintain mitochondrial structure and function stable. When mitochondrial oxidative respiratory chain dysfunction, leading to excessive accumulation of oxygen free radicals, it’s easy to form oxidative stress, which is considered to be an important cause of neurodegenerative diseases, cancer and many other chronic diseases. Thanks to PHB stably regulate mitochondrial function is closely related with cellular oxidative stress, it’s gradually becoming a hotspot protein in researching cellular oxidative stress tolerance. However as a member of PHB family, no one has studied the relationship between PHB2 and the lampreys adaptability under oxidative stress environment. This paper focus on research of tissue and cell oxidative stress action and mechanism and of lampreys PHB2 proteins, which is helpful for revealing lamprey PHB2 protein function, at the same time providing clues for study mechanisms of human evolution PHB2 in oxidative stress.In this study, immunofluorescence assay indicated the position of lamprey Lm-PHB2 protein in gill and kidney cells, doxorubicin injection and H2O2-stimulated were used to construct lampreys oxidative stress animal models and human normal cells oxidative stress model, QPCR and western blot detected Lm-PHB2 distribution and expression of each organization in response to oxidative stress stimulation. With the addition of protein was incubated with cells and construct eukaryotic expression vector and cell transient transfection experiments, MTT was used to assay role of recombinant protein rLm-PHB2 and eukaryotic protein Lm-PHB2 in Chang liver cells and human lung epithelial cells under oxidative stress, and confocal microscopy was used to detect position and migration of Lm-PHB2 in human Chang liver cells under oxidative stress conditions, QPCR analyzed the relationship between of Lm-PHB2, VDAC3 and TP53 uder oxidative stress conditions. The results show that, Lm-PHB2 mainly localized in the mitochondria and nucleus; Lm-PHB2 were expressed in the main organization and tissue of lampreys, which is closely related to oxidative stress; rLm-PHB2 can increase oxidative stress tolerance of human Chang liver cells and lung epithelial cells; Lm-PHB2 enhance Chang liver cells from oxidative stress tolerance is closely related to the migration from nucleus to mitochondrial; Lm-PHB2, VDAC3 and TP53 expression showed a different change in different tissues under conditions of oxidative stress. In summary, lamprey Lm-PHB2 can enhance cellular oxidative stress tolerance in human Chang liver cells, which is likely to be achieved by positioning and migration of the protein, while in lamprey tissues and cells may be related to interactions of VDAC3 and TP53 through multiple signaling pathways.