Research Of Percutaneous Impulsive Current Stimulation Of Hepatic Region In Exercise-induced Fatigue Rats On The Function Of Hepatic Mitochondria

BackgroundExercise fatigue is common in military training problem, when after the occurrence of the exercise fatigue, if the fatigue of body not corrected in a timely manner and recovery, accumulating over a long period, due to "overwork", and lead to "chronic fatigue syndrome" sub healthy state of emergence, serious influence is often the military physical training. At present, taken by various anti fatigue measures, the effect more ineffective, how to use simple scientific and effective method to relieve sports fatigue, improving training results, and reduce the occurrence of sports injury is a subject of research value and the urgent demand. So, for how to delay the exercise fatigue development and how to speed up the sports fatigue recovery, is one of the hot topics in the studies of molecular biochemistry, military training of sports medicine and sports medicine. Through further study to analyze the pulse current of percutaneous stimulation the close relationship between sports fatigue rat liver and rat liver mitochondria respiratory enzymes activities, and enrich and improve the pulse current transcutaneous stimulation liver area of rats with exercise fatigue related series of research, as the "pulse current transcutaneous stimulator" devoted to the practical application to provide more enrich perfect theory basis.ObjectiveThrough exhaustive exercise of no-load swimming, we can establish an experimental model on exercise-induced fatigue animals. By observing the effects of percutaneous impulsive current stimulation of hepatic region on the hepatic mitochondria, respiratory chain enzyme complex activity, Na+-K+-ATP enzyme and Ca2+-Mg2+-ATP enzyme activity in exercise-induced fatigue rats and further discussing and describing the relationship between the percutaneous impulsive current stimulation of hepatic region and exercise-induced fatigue, and knowing the change of the main enzyme activity in mitochondria after the exercise-induced fatigue and the influence of percutaneous impulsive current stimulation on hepatic energy metabolism, we can improve the relevant mechanism about percutaneous impulsive current stimulation on relieving fatigue.MethodsEighty healthy eight-week-old Wistar male rats with 204±15g body mass which were provided by Experimental Animal Center of Henan Province were chosen. The feeding room temperature was 23±2℃.The humidity was 41±15%.Supplied with natural light, the rats which could eat and drink freely were fed adaptively for one week in separate cages. All of the animals did not do any swimming exercise before the experiment. The experimental animals were randomly divided into 4 groups of 18 each leaving the remaining 8 rats as spare:group A, group B, group C and group D. The rats of different groups were fed separately. During the first 3 days, each rat learned swimming for 30 minutes every day. The water temperature was 32±1℃, and the water depth was 70cm. Then the experiment was conducted for 5weeks in a row. Rats in the group B, group C, and group D received swimming training for 6 days every week and twice a day. Each time they needed to reach exhaustion. When the rats stopped and floated in water, they were forced to keep the state of motion with the help of a stick. If the rats did not break the surface of water 10 seconds after sinking, then they reached exhaustion. Each rat’s exhaustive swimming time was recorded. After swimming of each group, the water was changed. Ketamine Hydrochloride Injection 0.25g/kg was injected into the abdominal cavity to anaesthetize the rats of each group, with the group B and group D after swimming and the group C before swimming.1024Hz impulsive current stimulation was given to group C before swimming and group D after exhaustive swimming. The current intensity was 10mA. For each group it lasted 20min, with the period as 1s.Rats in each group were sacrificed in batches at the weekend of 1st,3rd, and 5th week. The animals going to be put to death were prohibited to eat and drink the night before. And they were put to death by decollation. Immediately after death,1g liver specimen was extracted by laparotomy. The specimen was immediately frozen by liquid nitrogen. After solving, it still kept the temperature of 2-8℃.After being cut into pieces by ophthalmic scissors, it was mixed with 9ml PBS liquid(PH7.4). The specimen then was fully homogenized by an ultrasound homogenizer. After 20 minutes’centrifuge (3000 turns/s), supernatant liquid was collected and packaged into different portions among which one portion was kept for detection and others for cryopreservation. The whole operating process was conducted in a temperature of 0-4℃. The hepatic mitochondria, respiratory chain enzyme complex(I-IV)activity of the rats,the activity level of Na+-K+-ATPase;Ca2+-Mg2+-ATPase were measured by spectro photometry and the protein quantification of the hepatic mitochondria in rats was measured by Bradfood protein quantification in each group at 1st,3rd and 5th week.Results1.Observation of the exhausting time:At the weekend of 1st week, the difference among the three groups in exhaustive swimming time was not statistically significant(P>0.05); at the weekend of 3rd week, the exhausting time of fatigue group (group B)was less than that of stimulating group before fatigue (group C) and stimulating group after fatigue (group D), and the difference between stimulating group before fatigue (group C) and stimulating group after fatigue (group D) in swimming exhausting time was no statistically significant(P>0.05), but the difference among the three groups in swimming exhausting time was statistically significant(P<0.05); at the weekend of 5th week, the exhausting time of fatigue group was less than that of stimulating group before fatigue and stimulating group after fatigue, and the difference between the exhausting time of fatigue group (group B) and stimulating group before fatigue (group C) in swimming exhausting time was no statistically significant(P>0.05), but the difference among the three groups in swimming exhausting time was statistically significant.2.The comparison of the hepatic mitochondrial respiratory chain enzyme complex(Ⅰ～Ⅳ)activity of the rats:At the weekend of 1st week, the difference among the four groups in terms of hepatic mitochondrial respiratory chain, enzyme complex(Ⅰ～Ⅳ)activity of the rats was not statistically significant(P>0.05); the level of activity of hepatic mitochondria, respiratory chain enzyme complex(I-IV)in control group, stimulating group before fatigue and stimulating group after fatigue in the weekend of 3rd week and 5th week was higher than that in fatigue group and the difference was statistically significant(P<0.05). The level of activity of hepatic mitochondria, respiratory chain enzyme complex(I-IV)in stimulating group after fatigue in the weekend of 3rd week and 5th week was higher than that in stimulating group before fatigue and the difference was statistically significant(P<0.05).3.The comparison between Na+-K+-ATP enzyme activity and Ca2+-Mg2+-ATP enzyme activity in hepatic mitochondria:At the weekend of 1st week, the difference among the four groups in terms of hepatic mitochondrial Na+-K+-ATP enzyme activity and Ca2+-Mg2+-ATP enzyme activity of the rats was not statistically significant(P>0.05); the level of activity of hepatic mitochondrial Na+-K+-ATP enzyme and Ca2+-Mg2+-ATP enzyme in control group, stimulating group before fatigue and stimulating group after fatigue in the weekend of 3rd week and 5th week was higher than that in fatigue group and the difference was statistically significant(P<0.05). The level of activity of hepatic mitochondrial Na+-K+-ATP enzyme and Ca2+-Mg2+-ATP enzyme in stimulating group after fatigue in the weekend of 3rd week and 5th week was higher than that in stimulating group before fatigue and the difference was statistically significant(P<0.05).Conclusions1.The exercise-induced fatigue can reduce the level of activity of hepatic mitochondria, respiratory chain, enzyme complex(Ⅰ～Ⅳ), Na+-K+-ATP enzyme and Ca2+-Mg2+-ATP enzyme, which can lead to decline of the exercise score.2. Percutaneous pulsive current stimulation of hepatic region in exercise-induced fatigue rats may improve the activity of the oxidant respiratory chain enzyme complex (Ⅰ～Ⅳ), promote the level of mitochondrial oxidative phosphorylation, which can increase ATP synthesis, promote exercise endurance and score, and relieve the exercise-induced fatigue.3.Percutaneous pulsive current stimulation of hepatic region in exercise-induced fatigue rats may improve the activity of Na+-K+-ATP enzyme and Ca2+-Mg2+-ATP enzyme, reduce the concentration of free calcium and calcium overloading in hepatic mitochondria, stimulate oxidative phosphorylation, accelerate the rate of respiratory chain and the elimination of exercise-induced fatigue.