| ObjectiveFANGZHENGXIANGYING is the theory about the relationship between prescript:ion and syndrome,emp hasizing the adaptation of the prescription to the syndrome.It is an important principle and method to guide clinical medication,and has lively features of traditional Chinese medicine.And YIFANGCEZHENG is one of the important methods of FANGZHENGXIANGYING,through the establishment of complex animal model,it carries out the corresponding experimental study of pesticide effect.It is not only intensive study of FANGZHENGXIANGYING,but also an exploration of syndrome model and the essence of syndrome.Studies have shown that YINXU,QIXU,XUEYU and FEIQIBUXUAN are all common syndromes at the time of sports fatigue,so we take it as the breakthrough point to explore YIFANGCEZHENG in sports fatigue.In this experiment,three adding and subtracting formulas were established on the basis ofthe Chinese medicine protection experience and clinical accumulation of the National short track speed skating team.The model of exercise fatigue induced by the combined stress stimulation of running and swimming in mice was selected to study the changes of the relevant pharmacodynamic indicators in four formulas during exercise fatigue,and to observe the differences of the corresponding syndromes,In order to explore and verify the method of YIFANGCEZHENG.Methods110 SPF grade Kunming mice were fed foradaptive 3 days.They were randomly divided into 11 groups according to the random number table method,eachgroup 10,weighed,recorded.Groips:normal group(N),model group(M),positive testosterone group(T),Yiguanjian high dose group(YG),Yiguanjian low dose group(YD),and high dose I group(1G),low dose 1 group(1D),high dose 2 group(2G),low dose 2 group(2D),highdose 3 group(3G)and low dose 3 group(3D).The normal group was fed normally,The rest of the mice started the experiment by running 15min and then immediately swimming 30min for 5 days.After gavage of 1h,Continue the above training,at the beginning of the sixth day Until the experiment is over.Condition:running speed:25 r/min,swimming temperature:22 ± 10C.On the sixth day of experiment,mice in each group were weighed.The normal group and model group were given equal distilled water.The other groups were given the medicine according to the weight,Conversion of the dosage according to the body surface area.YG group 20.80 g/K g/d,YD group 8.32 g/Kg/d,1G group 27.56 g/Kg/d,1D group 11.02 g/Kg/d,2Ggroup 25.33 g/Kg/d,2D group 10.13 g/Kg/d,3G group 22.10 g/Kg/d,3D group 8.84 g/Kg/d。T group 41.6 mg/Kg/d.On the 15th day of the experiment,mice were randomly selected to gavage and exercise for 1h or so.The blood collection from the orbit was about 0.6ml-1ml.After the blood was taken,the mice were decapitated and the cerebral cortex(frontal lobe and parietal lobe,>30mg)and hippocampus(>20mg)tissues were separated under ice bath.The blood samples were centrifuged(40℃ 15min,10000r/min),and the serum was absorbed.The cortical and hippocampal tissues were weighed,and the samples were stored at-78 ℃ cryogenic refrigerator.CK,BUN,T and C were quantitatively determined by ELISA.Main experimental steps:preparation of reagents,sample and standard products→fill sample(96 hole plate)→ adding enzyme,heating→washing,adding chromo genic agent→terminating color→immediately select 450nm condition to measure OD.by ELISA instrument.Quantitative analysis of MAO central neurotransmitters in the cerebral cortex and hippocampus of mice by the Coularray-EDC Ⅱ HPLC instrument of ESA company of the United States.The steps are shown in the text.The statistical data were analyzed by SPSS 22.0.The data were expressed by X±S.The data were compared by single factor variance analysis and Q test.inspection level a=0.05、0.01.Results1 Changes in weight of mice in each groupWeight change can objectively reflect the fatigue state of the animal body and is an important index to evaluate the fatigue degree of exercise fatigue.The results show that:There was no significant difference in preexercise weight between N,M and T groups.After 5 days of running and swimming exercise,N group did not exercise,the weight of the most significant increase;There was no significant difference in weight between M and T groups,but both of them lost weight.On the tenth day of administration the weight of group N continued to rise,and the weight of the three groups was the heaviest.The weight of group M slowly increased and the weight was the lightest.The difference between group N and group N was significant(P<0· 05);the weight of T Group continued to rise,which was larger than that ofM group,lower than that ofN group.2 Results of Determination of serum indexesThe M group compared with N group CK content increased,the T group compared to the M group significantly decreased(P<0.05),consistent with the literature report;3D group compared to the T group reduced CK content effect,consistent with the role of testosterone group(P<0.05).Compared with the N group,the BUN content in the M group increased significantly(P<0.01),and the T group had a significant decrease compared with the M group(P<0.01),indicating that the model animals were in fatigue state,the deconposition of muscle protein was accelerated,and the excretion slowed down.The 3G and 3D groups had the same effect with testosterone group.Compared with the M group,they all showed different degrees of reducing the effect of BUN(P<0.05),but all the YG、YD、1G and 1 Dgroup had no reduction effect,and there was no dose effect relationship.The testosterone content of the body decreased after the fatigue caused by the high amount of exercise.The level of blood testosterone in the M group was significantly lower than that in the N group(P<0.05),and the blood testosterone in T group increased significantly after oral testosterone(P<0.05).All of the fomula had the effect of maintaining T content,which was close to normal content,and increased significantly in relation to the M group,except for the YD group and the 1G group.The 2G、2D、3G and 3Dgroup had the effect of maintaining testosterone level(P<0.01 or P<0.05),and the high and low dose relationship was shownExercise stimulates the C value increased(compared with the N group)T group than the M grotup,C content was higher,compared with the N group increased significantly(P<0.05),with exercise stress and excessive compensatory performance;the 3G group and 3D group increased significantly(compared withthe M grovp,P<0.01)1Gand 1D group showed decreased(compared with the M group).The T/C ratio reflects the effect of body hormone level on physical ability in sportsfatigue.The T/C ratio of the M group was significantly lower than that of the N group(P<0.05),which proved that the exercise stress fatigue model was consistent with the literature.1G group and 1D group were significantly higher than that in the M group(P<0.05).3 Results of determination of central fatigue transmitterThe NE content in the mouse cerebral cortex was significantly higher than that in the normal group(P<0.05),which showed that the central neurotransmitter of the body was active when sports fatigue occurred.The NE content of T group was significantly lower than that of the model group(P<0.01).The content ofNE in the cortex of each group was consistent with that of T group,but the performance of each group was different.Content of DA、DOPAC and HVA in mouse cerebral cortex.Compared with the N group,the content of DA decreased significantly(P<0.05),the T group increased significantly,and the value was the same with N group(P<0.05).Compared with the M group,there were significant differences(P<0.01,P<0.05)between the 2G and 2D groups,but there was no significant difference between the two groups.The metabolic products DOPAC and HVA were similar to DA.Because 5-HT is an inhibitory neurotransmitter,its effect is closely related to the location and different state ofbody,so the content of5-HT in cerebral cortex is varied.The experimental results showed that the M group had a decreasing trend,and the T group had a significant increase(P<0.05).Both the YG and the YD group were reduced.1G and 1D group performance was consistent with T group,and the 5-HT level increased;The change of the 2G and 2Dgroups is not obvious.There were no statistical differences between the 3 G and 3 D grops.The content of the metabolite 5-HIA is basically the same as that of 5-HT.The fnction of hippocampal neurons is closely related to memory,learning and part of emotional activity,but the mechanism of change during fatigue is not clear.The experimental M group was significantly higher than the N group(P<0.01).The contents of NE in the T group increased as well as in the M group,which indicated that the related neural activity increased during sports fatigue.The contents of NE(compared with the N group)were consistently increased in all the groups,but the contents were different and no statistical difference.The content of DA,DOPAC and HVA in the hippocampus of mice.The DA level of the N group was higher than that of the M group,which was significantly different from that of the T group(P<0.05),indicating that sports fatigue also caused the change of DA,but testosterone had no direct effect on DA content in hippocampus of sports fatigue(P>0.05).YG、YD、1G、1D、2G and 2D groups were all lower than those in the M group,the difference was significant(P<0.05).However,the 3G、3D groups all showed the increase of DA level,and the 3D group was more obvious(P<0.01).If the changes in the content of DOPAC and HVA are taken into consideration,the uncertainty of the DA neuro transmitter can be analyzed.According to data,hippocampal 5-HT level in M group was significantly lower than that in N group(P<0.01),T group was also significantly lower than that in N group,and there was no difference between Ml group and T group,indicating testosterone had no effect on 5-HT content during sports fatigue(P>0.05).The 3G and 3D groups showed increased 5-HT content,similar to normal group(P<0.01);There are no significant difference YG YD group,1G、1D group,2G、2D group with the M group 5-HT level,but the combination of metabolite levels of 5-HIAA,there are mechanisms influencing the metabolic rate,different potential ways.Conclusion:1 In this study,testosterone was used as the contrast of the study.It was reasonable and observed that it affected the regulation of central neurotransmitter secretio-2 Both Yi-Guan-Jian and changing prescription showed different anti-fatigue effects,which showed that the study of new anti-fatigue drugs or health products based on the Yi-Guan-Jian had a solid foundation3 Combined with the main syndrome and rules of treatment of four formulas,the comparativte analysis shows that they have obvious differences in pharmacodynamic characteristics,along with the different principles and treatment,reflecting different emphasis on anti-fatigue effects and different regulation mechanisms.Reflects under the same pharmacodynamics model,different main syndromes of the objective degree.4 This paper provides an example of the research experiment of Traditional Chinese medical Although the experiment is not enough,it can be used as a way to deepen the theoretical research of formula corresponding to syndromes. |
PDF Full Text Request