| Fatigue is the result of the combined effects by multiple factors. The status of fatigue,drowsiness, weakness, headache, insomnia, etc. which resulted from long time exercise oremotional labour usually belongs to physiological fatigue, and is thought to be protectivesignals issued by the body. These phenomena don’t hurt the body and can be restoredthrough rest and recuperation. However, over-fatigue caused of the accumulation of fatiguecan affect the function of the body and cause organ lesions that have serious harm tohuman health. Fatigue crowd is expanding with compact social rhythm, busy work and life.Moreover, for special populations such as athletes with frequent high intensity training,workers under hypoxia, high temperature, and high humidity environment as well as closedenvironment for long time, officers accepting long-term military training and so on, fatiguecontinue to produce but can’t recover in time resulting in accumulation, which caused greatdamage. Therefore, to find effective drugs to delay the occurrence of fatigue or quicklyeliminate fatigue are of great significance for social medicine, sports medicine, andmilitary medicine.At present, the clinical medicine for fatigue is mostly chemical drug with the functionof being excited central. These drugs have a significant effect, but also have large sideeffects especially for the central nervous system, and most have addiction and dependence.Relatively speaking, due to multiple medicinal ingredients, multiple targets and ease role,Chinese medicine and its compound preparations attracted wide attention recently.Compound Acanthopanacis Senticosi Radix consisting of Acanthopanax senticosus,Astragalus membranaceus and Changium smyrnioides have been used for treating fatigueas a hospital preparation in clinic with significant effect, and we carried out a systematicstudy on the anti-fatigue of Acanthopanax senticosus in previous, thus the project studiesthe above compound, in order to lay the foundation for the development of anti-fatiguedrugs.In this work, we optimized and validated the ratio of compound by Uniform Designcombined with weight-loaded swimming test, and swimming exhaustion time wasregarded as index to evaluate the anti-fatigue effect. The experimental data was analyzedby Uniform Design U3.00, and obtained the regression equation Y=8.46+0.184X1+0.739X2-0.194X3. The equation was significant and further optimizationas well as validation experiments showed that the compound with the ratio3:2:1(Acanthopanax senticosus: Astragalus membranaceus: Changium smyrnioides) are the bestchoice with obvious anti-fatigue. We used the weight-loaded swimming experiment withmice to study the anti-fatigue effect of different extractions and found water extraction andalcohol precipitation method has better effect. The extract was separated and purified byD101macroporous resin, and then each elution fraction was to mice. The swimmingexhaustion time and the levels of BUN, TG, LDH and CK in serum were regarded as indexto evaluate the anti-fatigue effect of different parts in weight-loaded swimming test. As aresult,30%and50%-70%fractions showed obvious anti-fatigue effect by enhancing theability of body to adapt to load, clearing up BUN quickly, enhancing the activity of LDH,removing lactic acid in serum quickly, decreasing the levels of TG, and pretecting themuscle cell membrane integrity. The constituents’ databases of three Chinese traditionalmedicines were established through searching literatures, and the chemical compositions ofeffective fractions were analyzed by UHPLC-Q-TOF-MS. The matching results showedthose30%ethanol eluates mainly included lignans and its glycosides from AcanthopanacisSenticosus, while50%-70%ethanol eluates mainly contained saponin compounds fromAstragalus membranaceus. In addition, the two fractions all included flavonoids. It can beconcluded that lignans, saponins and flavonoids are the active ingredients of thecompound.Then, we study the effect of two fractions on fatigue produced under closedcompartment with high temperature, high humidity, nosie, ionizing radiation and so on.The changes of cell ultramicrostructure in hippocampus, thymus, testis and small intestinewere observed by Transmission Electron Microscope. The result showed that apoptosisphenomena were obvious in different tissues of mice in model group, while the apoptosisphenomena were improved both in groups gavaged with30%and50%-70%ethanoleluates. Compared with30%ethanol eluates,50%-70%ethanol eluates have moresignificant improvement. The morphologies of neurons, lymphocytes, intestinal villi andepithelial cells were closed to normal. However, the ameliorative effect on apoptosis intestis is not obvious. It can be concluded that50%-70%ethanol eluates could significantlyrelieve fatigue caused by multiple factors in special circumstances by inhibiting apoptosis,protecting hippocampus, thymus and intestinal tissues from injury and promoting the repair after injury. In acute toxicity test, total extract and30%ethanol eluates were gavaged bymaximum volume and peak concentration, and there was no significant acute toxicity.Meanwhile,50%-70%ethanol eluates also had no obvious acute toxicity at the dose ofMTD. The doses are equivalent to301,943and685times of clinical dose, so the clinicaltherapeutic dose is safe.The experiment defined the best ratio and extraction method with pharmacologicalactivity as a guide. The30%and50%-70%ethanol eluates were the effective fractions forfatigue caused by continuous movement and confirmed that lignans, saponins andflavonoids are the main active ingredient. In addition, we found that50%-70%ethanoleluates have good effect on fatigue produced under special conditions, and have no toxicreaction. Therefore, the compound Acanthopanacis Senticosi Radix is worth to bedeveloped. |
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