The Research On The Relationship Between Mitochondrial Biogenesis And Inflammatory Response By Electrical Stimulation In C2C12 Cells

Objective: At present,it is not clear that the relationship between exercise-induced mitochondrial biogenesis and inflammation.To observe the regulation mechanism of mitochondrial biogenesis and inflammatory factors,this study simulated aerobic endurance exercise by electrical stimulation of medium intensity in C2C12 cells,and gene silencing PGC-1? to observe the mechanism between mitochondrial biogenesis and myokines.Methods: After C2C12 cells were differentiated into myotubes,here we applied to intervene electrical stimulation and PGC-1? gene silencing in two ways.The groups were divided into control group(Group C),electrical stimulation group(E),PGC-1? gene silencing group(Group R)and PGC-1? gene silencing plus electrical stimulation group(RE).RT-q PCR technology for silencing group of cells,ROS levels were detected by fluorospectrophotometer and laser scanning confocal microscope,mitochondrial membrane potential changes were detected by JC-1 assay kit,mitochondrial respiratory function tests were detected by Oxygraph-2k,cell proliferation and cell toxicity were detected by MTT assay kit,and Western blot detection of PGC-1?,NF-k B,Nrf2,TNF-?,IL-6,COXIV protein expression.Results: 1.In the E group,ROS production were decreased,while mitochondrial membrane potential were increased,oxygen consumption rate was increased and cell respiratory function was improvement.Expression of PGC-1??Nrf2? NF-k B and COXIV expression were increased,while IL-6 and TNF-? basal levers significantly lower than the control group.2.When PGC-1? expression was inhibited,ROS production were increased,mitochondrial membrane potential were decreased,cell respiratory function and oxygen consumption rate were decreased,NF-k B expression were decreased slightly,IL-6 and TNF-? expression rise,COXIV expression was lower than control group protein expression.3.In the RE group,ROS production were increased,but not as obvious as R group,mitochondrial membrane potential were increased,not as obvious as E group.Compared with the R group,there was no difference in cell respiration function.The expression of PGC-1? was lower than E group but higher than R group,which no difference in C group.Compared with E group,the expression of NF-k B protein decreased,and no difference in R group.Nrf2 protein expression were increased,there were significantly different between E group and R group.IL-6 protein and TNF-? protein expression were significantly increased.The expression of COXIV protein was not significantly different in R group.Conclusion: 1.In the moderate intensity endurance training by simulation of electrical stimulation,on the one hand,which can up regulate the PGC-1?expression and improve mitochondrial biogenesis,on the other hand,it can inhibited the inflammation of the IL-6 and TNF-? production,which have a good effect on inflammation and slow down the occurrence of chronic inflammation.2.Though RNA interference reduced mitochondrial biogenesis,IL-6 and TNF-? expression was not significantly increased in the R group but significantly increased in the RE group,which shows that exercise can play a critical role in muscle inflammation,from the side that mitochondrial biogenesis might be related to the level of inflammation.