Mechanism Of Action Of Shengyang Sanhuo Decoction In The Treatment Of Chronic Fatigue Syndrome

Research backgroundChronic fatigue syndrome is a group of complex symptoms.Patients have intestinal flora structure and function disorder,metabolic dysfunction,immune dysfunction,cognitive impairment and other pathological manifestations.With the acceleration of social rhythm,the incidence rate of chronic fatigue syndrome is also increasing,and the prevalence rate of the whole world is 3.5%.CFS is a gradual process,the cause of the disease has not been clear,but it is mostly related to work stress,overwork,lack of sleep and psychological stress.With the deepening of scientific research,most scholars can accept the view that chronic fatigue syndrome is caused by a variety of complex factors,and most patients are accompanied with different degrees of neuropsychiatric symptoms.As the energy factory of cells,mitochondria are closely related to energy metabolism.In recent years,many studies have confirmed that intestinal flora,as the second genome of human body,is not only directly involved in the normal physiological and metabolic activities of human body,but also closely related to the occurrence and development of chronic fatigue syndrome.Sheng yang san huo tang is a classic prescription,which originated from the chapter of"stomach qi due to heat injury" in LI Dongyuan's differentiation of internal and external injuries.It is a masterpiece of "fire stagnation".The combination of Qi tonifying medicine and wind medicine can not only supplement without stagnation,but also disperse without injury.It is considered to be the representative prescription of "ascending Yang and dispersing fire" in traditional Chinese medicine.According to the research idea of "according to the pathogenesis,according to the law",Sheng yang san huo tang can be divided into two groups,one is the tonifying deficiency group(ginseng,white peony,raw licorice,fried licorice);the other is the Sheng yang San yu group(Shengma,Gegen,bupleurum,notopterygium,Fangfeng,Duhuo).At present,some clinical studies have shown that Sheng yang san huo tang can improve the fatigue symptoms of patients with chronic fatigue syndrome,but the mechanism of Sheng yang san huo Tang in the treatment of chronic fatigue syndrome is not clear,and the specific mechanism of its anti fatigue and the role of different prescriptions under the compatibility principle of "Sheng yang sanh uo method" are still worthy of further discussion.Therefore,the therapeutic effect and mechanism of Sheng yang san huo Tang on chronic fatigue syndrome were studied.research objectiveThis study combined with the network pharmacology research method,studied the mechanism of Sheng yang san huo tang and its removing prescription in the treatment of chronic fatigue syndrome.research contentsThis subject is divided into two parts:network pharmacology prediction experiment,pharmacodynamic evaluation and animal mechanism exploration.The active components of Sheng yang sanh uotang were extracted by tcmsp database.The official gene name of the target target was corrected using the UniProt database.The aim of chronic fatigue syndrome was obtained by OMIM,TTD,Gen eCards and dispenet gene database,and the effective target of the treatment of chronic fatigue syndrome was obtained.The above targets were input into string database to predict protein protein interaction network,and the enrichment analysis of go and KEGG was carried out by David database.In order to evaluate the effect of Sheng yang san huo tang on chronic fatigue model mice,the changes of behavior,morphology of skeletal muscle and myocardial cells,and ultrastructure of mitochondria were observed.Furthermore,the possible mechanism was explored,and the effects of drugs on AMPK/PGC-1?signaling pathway,mitochondrial dynamics related proteins,inflammatory factors,serum metabonomics and intestinal flora were observed.research resultsNetwork pharmacology prediction results show that Sheng yang san huo tang has 128 active components with anti fatigue effect,involving 274 targets,of which 65 genes share the same target with diseases,mainly enriched in 145 biological processes and 41 signaling pathways,playing a role in the treatment of chronic fatigue syndrome.STAT3,AKT1,TNF,mapk1,TP53,VEGFA,EGFR,IL1B,CCND1,myc were predicted.These 10 target proteins may play an important role in the anti fatigue process of Sheng yang san huo tang.The main antifatigue active component is flavonoids,which has the characteristics of "multi-target,multi-channel,collaborative" anti chronic fatigue.In vivo pharmacodynamic evaluation experiment,behavioral experiment results showed that Sheng yang san huo tang and its disassembled formula could not increase the weight of chronic fatigue mice.The exhaustive swimming test showed that the whole formula ofSheng yang san huo tang and its disassembled formula could prolong the swimming time of mice.In addition,in the short term(15 days of administration),the exhaustive swimming time of mice in Buxu group was longer and the effect was better.The effect of Sheng yang san huo tang is better for long-term(30 days)chronic fatigue.The pathological results showed that the mitochondria of skeletal muscle and myocardium of chronic fatigue mice were seriously damaged and difficult to recover.Sheng yang san huo tang and its disassembled formula could better protect the stability of mitochondrial membrane structure of skeletal muscle and myocardium.ELISA results also showed that Sheng yang san huo tang and its disassembled formula could effectively reduce the serum inflammatory factors of chronic fatigue mice and further affect the mitochondria Sheng yang san huo tang has the best effect on body energy metabolism.Western blot results showed that Sheng yang san huo tang could significantly up regulate the protein expression of AMPK and PGC-1?,up regulate the protein expression of mitochondrial fusion related proteins OPA1,mfnl and mitochondrial division related protein drp1.It is suggested that chronic fatigue can inhibit the energy metabolism disorder caused by abnormal mitochondrial function in mice,indicating that Sheng yang san huo tang and its disassembled formula can affect mitochondrial division and fusion,and enhance the function of mitochondria to meet the body's energy demand.ELISA results showed that Sheng yang san huo tang and its components could significantly reduce IL-6.According to literature studies,IL-6 could up regulate the expression of FIS1 and down regulate PGC-1,which made the mitochondrial dynamic balance tend to split.It is concluded that Sheng yang san huo tang and its disassembled prescriptions may affect mitochondrial division by reducing the expression of IL-6 inflammatory factors.The results of intestinal flora showed that the diversity and species richness of intestinal flora of mice in Sheng yang san huo tang and its dismantling groups were significantly increased,among which the whole recipe of Sheng yang san huo tang had the most obvious effect.Sheng yang san huo tang and its removing prescription can have effect on 4 species of bacteria and 25 species of bacteria.By increasing the probiotics ratio of chronic fatigue mice,the inflammatory response of the body will be affected and energy metabolism will be affected.The results of NMR showed that chronic fatigue could lead to the change of metabolites formic acid,a glucose,?glucose,lactate,creatine and phosphorylated creatine,citrate,acetate,valine,isoleucine,leucine and dimethylamine.Sheng yang san huo tang and its removing prescription can interfere with it in different degrees and affect the metabolism of sugar and amino acids.Joint research showsAt the genus level,Lachnospiraceae_NK4A136_group is strongly positively related to lactate,and atoposides is strongly positively related to lactatea glucose,alanine,NAG and acetate was negatively correlated with the Jeotgalicoccus.Conclusion(1)The main antifatigue active components of Sheng yang san huo tang are flavonoids.The antifatigue pathway involves inflammation,immunity,energy metabolism and other pathways,involving multi protein targets.(2)Sheng yang san huo tang and its disassembled formula can improve the behavior of model mice,increase the exhaustive swimming time of model mice,shorten the tail suspension static time,and the effect of Buxu group is better after 15 days of administration.After 30 days of administration,the effect of Sheng yang san huo tang was better.(3)Sheng yang san huo tang may further regulate mitochondrial division and fusion by activating AMPK/PGC-1? signal.(4)Sheng yang san huo tang and its disassembled formula can regulate mitochondrial division and fusion,protect skeletal muscle and myocardial mitochondrial structure and improve mitochondrial function by affecting inflammatory factors IL-6,mitochondrial fusion proteins OPA1,mfnl and mitochondrial mitotic protein drp1.(5)Sheng yang san huo tang and its disassembled formula can change fecal flora and small molecule serum metabolites of fatigue mice in varying degrees,enhance immunity,protect mitochondrial function,and play the role of anti fatigue and anti depression.In conclusionSheng yang san huo tang and its removing prescription can enhance the immunity of the body,protect the mitochondrial function,and play the anti fatigue and antidepressant functions.In the above process,the fecal bacteria group and small molecule serum metabolites of mice changed.In conclusion,Sheng yang san huo tang may have an effect on mitochondrial dynamics through a variety of ways,improve energy metabolism and alleviate fatigue and depression symptoms.