龟鹿益神方改善慢性疲劳综合征作用机制的网络药理学分析与验证OA

Aim:To analyze and verify the mechanism of Guilu Yishen Formula in improving chronic fatigue syndrom(CFS)via network pharmacology.Methods:Network pharmacology was applied to predict the main active components and core targets of Guilu Yishen Formula for CFS treatment,followed by molecular docking verification.Thirty SD rats were suc-cessfully established as CFS models and randomly divided into the model group(distilled water),positive control group(tri-sodium fructose,0.09 g/kg),and Guilu Yishen group(5.85 g/kg Guilu Yishen Formula decoction),with 10 rats in each group.Intragastric administration was started on day 7 after modeling,once daily for 14 consecutive days.Another 10 normal rats were set as the normal control group.After the last administration,the exhaustive time was measured in each group.Blood samples were collected from the abdominal aorta to detect serum levels of creatine kinase(CK),lactate dehydrogenase(LDH),5-hydroxytryptamine(5-HT)and dopamine(DA).Skeletal muscle tissues were harvested for pathological observa-tion under light microscope.Immunohistochemistry and PCR were used to detect the protein and mRNA expression levels of monoamine oxidase A(MAOA),monoamine oxidase B(MAOB),5-hydroxytryptamine receptor 2 A(HTR2A)and 5-hydroxytryptamine receptor 3A(HTR3A).Results:Network pharmacology and molecular docking results indicated that the main active components of Guilu Yishen Formula against CFS were 3-[3-hydroxymethylene tanshinone,tenuifolin A,luteolin,cycloartenol and ellagic acid,and the core targets were MAOA,MAOB,HTR2A and HTR3A.Compared with the normal con-trol group,the model group presented shortened exhaustive time,elevated serum CK and LDH levels,decreased serum 5-HT and DA levels(P＜0.05),skeletal muscle fiber degeneration and partial loss of muscle fiber structure.The mRNA and pro-tein expressions of MAOA and MAOB in skeletal muscle were increased,while those of HTR2A and HTR3A were decreased(P＜0.05).Compared with the model group,the Guilu Yishen group showed alleviated skeletal muscle fiber degeneration without loss of muscle fiber structure,and all the indicators were improved(P＜0.05).Conclusion:Guilu Yishen Formula may ameliorate CFS by down-regulating MAOA and MAOB and up-regulating HTR2A and HTR3A in skeletal muscle,there-by modulating the 5HT-ergic synaptic pathway.