Study On Anti-fatigue Effect Of Maca Compound Preparation And Its Probable Mechanism

Maca is a traditional medicinal and edible plant with significant anti-fatigue effects.In this study,Maca Aqueous Extract（ME）was used as the main raw material.According to the concept of“holistic treatment”in traditional Chinese medicine,7 kinds of“medicinal and food homologous”plants such as Polygonatum and Astragalus,were selected.The are compatible with each other in the Maca compound preparation.The compound was optimized by orthogonal design experiment,and the compatibility effect of its water extract（CE,Compound Aqueous Extract）was studied.Based on this,a compound oral liquid with significant anti-fatigue effect was developed,and its possible mechanism of action was explored from the perspective of network pharmacology.The contents of study are as follows:（1）Formula optimization of Maca compound preparation.Based on the theory of traditional Chinese medicine,the plants in the prescription are divided into 4 categories.The viability of C2C12 cells is used as an indicator to explore the optimal ratio of the two water extracts within each factor in the model of H₂O₂-induced oxidative stress cells.Negative prescriptions were set up for each factor,and the synergistic effect of the combination of various factors in the compound prescription was studied.The formula was optimized by L9（3⁴）orthogonal design,and the best ratio in Maca compound preparation was obtained.（2）Evaluation of anti-fatigue potential of Maca compound preparation in vitro.CE significantly inhibited the reduction of C2C12 skeletal muscle cell viability induced by H₂O₂,and reduced muscle cell damage and necrosis detected by Calcein-AM/PI double staining.CE concentration-dependently increased glycogen and adenosine triphosphate（ATP）content,and promoted energy metabolism in C2C12 cells.In addition,CE-induced activation of skeletal muscle energy metabolism might up-regulate the biogenesis and function of mitochondria,thereby alleviating the damage of skeletal muscle cells caused by oxidative stress.（3）Evaluation of anti-fatigue effect of Maca compound preparation in vivo.The Maca compound preparation improved the grip of mice forelimbs and the exercise time on the fatigue rotating rod instrument,and relieved the exercise fatigue caused by weight-loaded forced swimming test.On the one hand,Maca and its compound preparations could clear the accumulation of blood lactic acid（BLA）,blood urea nitrogen（BUN）,and reduced the activity of lactate dehydrogenase（LDH）,thereby promoting glycogen synthesis,ATP production capacity and reactive oxygen species（ROS）removal in skeletal muscle.By strengthening the muscle structure of mice,it protected skeletal muscle fibers from acute exercise damage,and promoted the recovery of exercise-induced peripheral fatigue.On the other hand,due to the significant antioxidant and anti-inflammatory effects,Maca and its compound preparations also regulated the brain fatigue.The central neurotransmitter of the tissue maintained the balance of excitability/inhibition of neurons and delayed the occurrence of central exercise-induced fatigue.（4）Prediction of the anti-fatigue mechanism of Maca compound preparation through network pharmacology.A total of 130 potential active ingredients in Maca compound preparations were screened by ADME parameters,of which 120 were anti-fatigue active ingredients,involving 116 anti-fatigue potential gene targets.A network of"single medicine-active ingredients-targets of action"was constructed.Through protein interaction network analysis,it is found that the proteins with greater degrees are:PIK3CA（Degree=34）,SRC（Degree=26）,HSP90AA1（Degree=18）,VEGFA（Degree=17）,PIK3CB（Degree=15）,indicating that these proteins might play an important anti-fatigue role in maca compound preparations.Biological process enrichment analysis was carried out through Gene Ontology（GO）,and it revealed that Maca compound preparation mainly regulated monoterpenoid metabolism,dopamine uptake in synaptic transmission,and phosphatidylinositol-3,4-diphosphate 5-Kinase activity and other pathways exert anti-fatigue effects.Based on the enrichment results of Kyoto Encyclopedia of Genes and Genomes（KEGG）metabolic pathways,it showed that maca compound preparations played an important role in TNF signaling pathway,phenylalanine metabolism,and AGEs in diabetic complications,RAGE signaling pathway and so on.