Study On The Interaction Between Antiproliferative Protein Prohibitin1 And ATP Synthase Under Exercise Fatigue

Objective:To study the possible interaction between PHB1 and F₀F₁-ATP synthase in the mitochondria of skeletal muscle,and the specific subunits of F₀F₁-ATP synthase.Methods:The animal treadmill was used in the experiment.80 healthy male SD rats?8 years?were perchased which were randomly divided into two groups after the first two adaptive feeding days,and their body weights and other relevant status were recorded?weigh before training every day;mental status observation;food intake:a day before training metering?.1.Acute exercise group:?1?Control group:20,regular feeding,without intervention.?2?Exhaustive exercise group:20,regular feeding,made the rats have exhaustive exercise and 8 weeks later;criteria for judging the state of exhaustion:the running form of rats turned from the formula stomp into prostrate type,no longer stuck in the end of the runway to continue running,and electrical stimulation cannot continue to drive rats running;2.Long-term exercise group:?1?Control group:20,regular feeding,without intervention.?2?Moderate intensity training group:20,6 training days a week,the first week in slope of 10%,run for10min at a speed of 10m/min;the second week in a gradient of 10%,speed up to15m/min for 10min more per day until the end of the second week,maintain this speed from the third week until the eight week;?3?Overtraining group:20,weekly training 6d,rest 1d,an incremental intensity treadmill exercise.From the first week of training,gradually increasing speed,slope and running duration.The treadmill speed started with 15m/min,then increase 5m/min per week,and increase no further at30m/min;slope started with 0°and then increase 5°per week until it was 15°;training duration started with 30min and increased 20min per week until it was110min.Gradually increased the number of weeks of training until 8 weeks.The animal models of skeletal muscle tissue samples were collected which were partly remained as samples and partly went through mitochondrial protein extraction kit to extract and isolate mitochondrial protein,then through Mass Spectrometry to find out the proteins combine with the PHB1 in F₀F₁-ATP synthase.Then the Co-Immunoprecipitation was adopted to verify the results of the Mass Spectrometry.Results:1. Mass spectrometry detected the subunits of ATP synthase,among four different movement patterns,finding the common proteins which combine with PHB1 in mitochondria of skeletal muscle:?and?subunit of ATP synthase.As the variety,exhaustive exercise group is the most,control group is the least.Compared with control group,moderate intensity training group had been detected with d subunit,overtraining group had been detected with B1 subunit.Compared with moderate intensity training group,overtraining group had been detected with B1 subunit,?subunit,O subunit;Compared with overtraining group.Compared with control group,exhaustive exercise group had been detected with?subunit2.FindingtheinteractionbetweenPHB1and?,?subunitsthrough co-immunoprecipitation.Conclusions:1. The interaction exists between PHB1 and F₀F₁-ATP synthase2. PHB1 interacts with?,?and subunits of F₀F₁-ATP synthase...