| [Objective]:The glucose metabolism of liver cell not only maintains the stability of the liver cells and blood glucose~'s internal environment,but also plays a vital role in maintaining the stability of sports performance.Only one endurance exercise training can cause significant changes in glycogen concentration in hepatocytes.In order to maintain stable internal environment of glycogen,this change will inevitably cause the liver cells to increase the glucose stress demand and transport large amounts of glucose to the liver cells,but research on liver cell glucose transporter has been relatively weak.So make sure the body respond to or adjust the exercise-induced stress events caused by the molecular mechanism of liver glycogen concentration changes for the understanding of the physiological function of liver and improving sports physical fitness is of great significance.whether the skeletal muscle or cardiac cells in the body,especially in the process of the movement of skeletal muscle contraction caused by its demand for glucose irritability,their intracellular glucose is regulated by GLUT4 protein.Although there are many members of the transporter family,only GLUT4 has the ability to transport glucose in stressful conditions.In the recovery stage of liver glycogen after vigorous exercise,the body also needs to transport a large amount of glucose to the liver cells in and the expression of GLUT4 in the HepG2 cells in the liver is found.Therefore,I infer that the glucose required by the exercise induced hepatocyte stress liver sugar recovery phase is done by GLUT4 and uses a mechanism similar to the skeletal muscle.This study was the first to give different glucose load stimulation conditions to simulate the large amplitude fluctuation environment of glucose in the body to explore the biological function of GLUT4 in the liver cells and the corresponding situation of liver glycogen and GLUT4 expression in the liver cells and look forward to finding the mechanism of regulating the GLUT4 in the liver cells through a single analog signal,"Caffeine induction."So as to provide a theoretical basis for promoting the rapid recovery of liver glycogen by means of corresponding means so as to maintain stable exercise performance.[method]:.Experiment 1(second parts)verified the stress characteristics of GLUT4 in hepatocytes.24 male SD rats(weight in 165-185g)were divided into 6 groups according to weight pair(control group 18h,fasting 18h group,high sugar 18h group,high sugar 42h group,low sugar 42h group,low sugar 42h and high sugar 24h group).In addition to the control group 18h,the remaining 5 groups of SD rats were trained for3 days,5%days of weight loading and 120min swimming training every day.After training,different diets was given and after the training of 18 h,h,66 h 42 batch materials,western blotting-biological detection technology is used to test each total protein in rat liver cells GLUT4 protein expression level,and by using sulfuric acid green ketone method to test different group's hepatic glycogen concentration.Experiment two(third part),using Caffeine induction,effectively avoided the effect of other signals on the experimental results,in order to study the mechanism of GLUT4gene regulation.The 24 male SD rats were divided into 4 groups of---C2 group,D2group,C24 group and D24 group according to the weight size"S"type.The liver tissues were taken respectively by 2H and 26h after the injection of liquid.The CaMII phosphorylation level in the liver cells was measured by C2 and D2 groups.[results]:(1)The total protein level of GLUT4 in 18h liver cells was 3.23 times as high as that of the high glucose 18h group(p=0.002<0.01);(2)The total GLUT4 protein level in the low sugar 42h group was 5.6 times as high as that of the high sugar 42group(p=0.001<0.01);(3)the total egg white level of the low sugar 42h group was 6.37times of the low sugar 42h after the high sugar 24h group(p=0.001<0.01);(4)high sugar liver fine.The concentration of glycogen was 2.31 times(p=0.004<0.01)in the low sugar 18h group;(5)the glycogen concentration in the liver cell of the high sugar42h group was 1.22 times(p=0.05)in the low sugar 42 group,and(6)the level of hepatocyte glycogen concentration in the 24h group with low sugar 42h was 1.3 times as much as that of the low sugar 42h group(p=0.05).The results showed that there was an inverse relationship between the total protein level of GLUT4 and the concentration of liver glycogen in each group,that is,when the concentration of liver glycogen was low,the expression of GLUT4 was high,while the expression of GLUT4 was low when the concentration of liver glycogen was higher.In experiment two,the total protein level of GLUT4 in C24h hepatocyte was 2.95 times as much as that in group D24(p=0.002<0.01),and the level of phosphorylation of CaMKII in C2h group was 4.58times(p=0.001<0.01)in D2h group.The results showed that the level of GLUT4 total protein and the level of phosphorylation of hepatocyte total protein and CaMKII in group C2h were much lower than that of group D24 and D2 in the control group,and all were statistically significant.[research conclusions]:(1)GLUT4 protein has the ability of stress transport similar to skeletal muscle and myocardium in liver cells.It can be responsible for the transport of glucose in stress state and play an important role in the rapid recovery of liver glycogen(2)sugar has a very strong modification effect on the expression of GLUT4 gene,and the change of glycogen concentration in hepatocytes may be the upstream signal.To regulate the expression of GLUT4 gene protein.(3)CaMK II signaling pathway plays an important role in regulating liver glucose metabolism,and the"Caffeine-Ca2+-CaMKII"signaling pathway exists in the regulation of GLUT4 protein expression in rat liver cells. |
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