| Objective:Based on spleen nourishing muscle theory,the efficacy and molecular mechanism of Jianpi Qiangji Decoction counteracting exercise fatigue were observed,and the pathogenesis of spleen deficiency in exercise fatigue and Chinese Medicine treatment to strengthen the spleen and benefit the qi in improving glycoisogenic resistance to exercise fatigue was elucidate.The MCT1+/-mouse model of exercise fatigue was established by using exhaustive swimming,and Jianpi Qiangji Decoction was running,and the relationship between its dose and size were investigated.The expression of molecular proteins and related genes of gluconeogenic metabolic pathway,CRTC2/CREB signaling pathway were examined to explore the mechanism of gluconeogenesis in liver to maintain the stability of exercise blood glucose,and to delay and avoid exercise fatigue.To investigate the mechanism of MCT1 involvement in gluconeogenic metabolism and analyze the mechanism of Jianpi Qiangji Decoction in exercise fatigue,it could further expand the application of Chinese Medicine in the field of sports medicine.MethodsMCT1+/-mice were randomly divided into control group(N=8),model group(N=8),low-dose group(JPQJD group,N=8),medium-dose group(JPQJZ group,N=8),and high-dose group(JPQJG group,N=8).The model group and JPQJG(low,medium,and high-dose groups)were given daily by gavage with saline and the corresponding drug concentration of JPQJZ.The model group and the JPQJG group(low,medium and high-dose groups)were received daily by swimming exhaustion with 5%body weight on a lead wire.2 hours before the exhaustion swimming exercise,saline and the corresponding drug concentration of JPQJG were administered by gavage in a volume of 0.15 ml once daily for eight weeks.The expression of lactate,blood glucose,MDA,SOD,GSH-PX,liver glycogen and myoglycogen in liver tissues,serum were measured by ELISA,and the expression of MCT1,PEPCK and G-6-Pase mRNA in liver tissues were detected by PCR.Western Blot method was used to detect the protein expression of CRTC2/CREB signaling pathway of liver tissues.Results1.Pathological morphological results:According to HE staining observation of gastrocnemius muscle morphology,exhaustive swimming caused pathological changes in gastrocnemius muscle and liver tissue.Control group had closely arranged muscle tissue structure,orderly arrangement of muscle fibers and clear structure of muscle membrane.Model group showed interstitial swelling,mild deformation of muscle fibers,with obvious inflammatory cell infiltration obviously.low,medium and high-dose groups of Jianpi Qiangji Decoction reduced the infiltration of inflammatory cells in the muscles of mice,resulting in more regular arrangement of skeletal muscle fibers and tighter cell arrangement.By observing the morphology of liver tissue and hepatocytes,the hepatocytes in the blank group were normal,intact,with clear hepatic cords,no infiltration,regular arrangement and clear intercellular boundaries.In the model group and low-dose group,the hepatocytes were intact,regularly arranged and slightly swollen.In the middle and high dose-groups,the morphology of hepatocytes was intact and regularly arranged,and swelling was not observed.2.Behavioral results:Compared with control group,model group successively showed rough and dull fur,no luster,yellowish fur,slight loss of fur on the back,which often bowed back in the cage,curled up and moved little,slow response to external frightening movements,or no response,no obvious struggle when grasping or even no struggle.Low-dose group mice had rougher and dull fur,and the phenomenon of fur loss was improved compared with the model group.The mice in low-dose group had rougher and darker fur,and its phenomenon of fur loss was improved compared with control group,while the mice in middle and high-dose groups had soft fur and normal behavioral activity.3.Exercise performance results:According to swimming exhaustion in mice,Jianpi Qiangji Decoction could prolong the time of swimming exhaustion.Compared with model group,low,middle and high-dose groups of Jianpi Qiangji Decoction could prolong the swimming exhaustion time of mice to different degrees,and their differences were statistically significant(P<0.05;P<0.01;P<0.001).Among which high-dose group of Jianpi Qiangji Decoction delayed the swimming exhaustion time more significantly,the differences were statistically significant(P<0.001).4.body weight and food intake result:There was no significant difference in body weight of mice before modeling.After eight weeks of swimming exhaustion training,the body weight of JPQJD group,JPQJZ group and JPQJG group increased significantly compared with model group,and the difference were statistically significant(P<0.05,P<0.001).According to the amount of mice’s food every week,compared with control group,the amount of food intake in model group decreased significantly,and the difference was statistically significant(P<0.01).The difference was statistically significant(P<0.01),and the amount of food intake in JPQJZ and JPQJG groups were significantly increased compared with model group(P<0.05).5.The content of lactic acid in liver tissue and serum,blood glucose content results:The lactic acid content in serum of mice,the lactic acid content in JPQJD group,JPQJZ group and JPQJG group were reduced compared with model group,and the differences were statistically significant(P<0.05,P<0.01).The lactic acid content in liver tissue of mice was detected,and the lactic acid content in liver of JPQJD group,JPQJZ group and JPQJG group was reduced,P<0.05,and the difference was statistically significant,and the lactic acid concentration in liver of mice was lower than that in serum.The blood glucose content in the serum of mice was measured,and compared with the model group,the blood glucose content in the serum of JPQJZ group and JPQJG group increased significantly,P<0.05,P<0.001,the difference was statistically significant.The blood glucose content in the serum of JPQJD group was elevated compared with the model group,P>0.05,but there was no significant difference.6.The content of liver glycogen and muscle glycogen results:Compared with control group,the content of liver glycogen and muscle glycogen in mice in model group was significantly decreased,P<0.001,and the differences were statistically significant.Compared with the model group,the content of liver glycogen and muscle glycogen in mice in the JPQJZ group and JPQJG group were significantly increased,P<0.001,the difference was statistically significant.7.The levels of MDA,SOD and GSH-PX in the serum of mice:Compared with model group,the levels of MDA in the serum of mice in JPQJD group,JPQJZ group and JPQJG group were significantly lower,(P<0.001),and the differences were statistically significant.The levels of SOD in the serum of mice in JPQJD group,JPQJZ group and JPQJG group were significantly higher compared with the model group,(P<0.01,P<0.001),and differences were statistically significant.The levels of GSH-PX in the serum of mice in JPQJD,JPQJZ and JPQJG groups were significantly higher compared with the model group,(P<0.05,P<0.001),and the differences were statistically.8.Expression of MCT1,PEPCK,and G-6-Pase mRNA in liver:Compared with control group,the levels of MCT1 mRA in liver were all increased,and although the levels of MCT1 mRNA in liver in model group were increased,P>0.05,and the differences were not statistically significant.Compared with model group,JPQJZ group and JPQJG group MCT1 mRNA levels in liver were significantly elevated,(P<0.05,P<0.001),and the differences were statistically significant.Compared with model group,PEPCK mRNA levels in JPQJZ group and JPQJG group were significantly elevated,(P<0.05,P<0.01),and the differences were statistically significant exhaustive swimming elevated G-6Pase mRNA levels were elevated.G-6-Pase mRNA levels in model group were higher than those in control group,P<0.05,and the differences were statistically significant.Compared with model group,G-6-Pase mRNA levels in model group were elevated compared with control group,P>0.05,bu tthe difference was not statistically significant.Compared with model group,level of G-6-Pase mRNA in JPQJZ and JPQJG group was significantly increased compared with that of model group,and the difference was statistically significant.9.Expression of CRTC2/CREB signaling pathway-related proteins:Compared with control group,CRTC2 and CREB in model group were elevated,P<0.05,and the differences were statistically significant.Compared with model group,the protein expression of CRTC2 and CREB were increased after the intervention of low,medium and high doses of Jianshuqiangmu Formula,and the difference was statistically significant P<0.05.Conclusion1.Jianpi Qiangji Decoction can effectively prolong the time of swimming exhaustion,enhance appetite,increase food intake and body weight in exercise fatigue mice,and slow down the damage of liver and gastrocnemius muscle caused by exhaustive swimming,thus enhancing the exercise ability of fatigue mice.2.Jianpi Qiangji Decoction can enhance the conversion rate of lactic acid in fatigue mice,increase blood glucose and enhance glycogen reserv.Exercise fatigue may also be related to oxidative stress.The MDA decreased and the SOD and GSH-PX content can be increased,by protecting the integrity of cell membrane,preventing lipid peroxidation,supplying energy to the body.It can protect the integrity of cell membrane,prevent lipid peroxidation,supply energy to the body and enhance the anti-fatigue ability of mice.3.The mechanism of Jianpi Qiangji Decoction improving anti-fatigue ability of mice may be through the upregulation of MCT1 activation of CRTC2/CREB pathway,and enhancing the expression of PEPCK and G-6-pase and accelerating the rate of gluconeogenesis,increasing expression of MCT1 accelerating the lactate transport to the liver,providing the original material for gluconeogenesis,increasing the lactate isomerization into glucose,supplying energy to the organism,thus delaying the production of fatigue to delaying exercise fatigue. |
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