Study On Anti-osteoporosis Mechanism Of Inflorescence Of Echinops Latifolius Based On Network Pharmacology

Objective The clinical practice of traditional Mongolian medicine has proved that Echinops latifolius has the pharmacological effects of osteoporosis,however,its active ingredient and mechanism are not clear yet.The main ingredients of Echinops latifolius were extracted and quantified based on the previous studies.In addition,the key active components,biological pathways and targets of Echinops latifolius in the treatment of osteoporosis were predicted using network pharmacology.Moreover,the pharmacodynamic effects and targets were verified,and the mechanisms of anti-osteoporosis function of Echinops latifolius were preliminary explore as well.Methods(1)The extraction of phenolic acids in Echinops latifolius was optimized by single factor test combined with orthogonal experiment with the content of chlorogenic acid and extract yield as index.(2)The total content of phenolic acids in Echinops latifolius were determined by ultraviolet spectrophotometry with content of chlorogenic acid as index.(3)The target bank of whole component in Echinops latifolius was established based on the chemical components of Echinops latifolius in CNKI and Pubmed,Swiss and Superpred databases,and previous research results of the our group.After the disease target banks built by searching OMIM database,compound target libraries and disease target libraries were compared to establish the correlative network model couple with Cytoscape software,and topology analysis of network model was carried out.The key active components,biological pathways and targets of Echinops latifolius in the treatment of osteoporosis were predicted by the DAVID online.(4)Replicated the op model of ovariectomized rats and the op model induced by D-galactose,divided into blank group,model group and drug-administered group.The drug-administered group included low,medium and high dose groups of Echinops latifolius and positive control group,the treatment lasted for 8 weeks.After the last treatment,samples were taken.The bone micro-structure was detected by CT;The serum bone metabolism index was determined by ELISA;The anti-osteoporosis targets HSD11B1,CYP191 and ALOX15 were verified by ELISA.Results(1)Extraction optimization results: Distilled water was used as extraction solvent,extraction time was 30 min,extracted two times,and the ratio of material to liquid was 1:12.(2)The total content of phenolic acid results: The average content of total phenolic acid in the drug was 61.33 mg/g according to ultraviolet spectrophotometry with content of chlorogenic acid as index.(3)The network pharmacological analysis results: The“ prescriptions—compound—targets-pathway network model” were including 28 compounds in Echinops latifolius and 371 targets,in which the key active components could be quercetin,apigenin,caffeic acid and n-hexadecanoic acid,and mainly regulating the expression of HSD11B1,CYP19A1,and ALOX15 in oxidation reduction pathway.(4)Pharmacodynamics test results: The level of serum E2 was increased,while the levels of serum DPD,TRAP,OC-BGP and PICP were decreased in middle and high dose groups compared with those in ovariectomized rats.Also,the levels of serum MDA and HYP were decreased and the level of SOD and TAOC were increased in low,middle and high dose groups than those of osteoporosis model rats induced by D-galactose,while the levels of ALP were decreased in the middle and high dose groups,which indicated that Echinops latifolius could increase the antioxidant capacity.At the same time,the connection of trabecular bone becomes better,and the cavity without bone connection is restored,which implied that Echinops latifolius could improve the pathological changes of bone microstructure of two kinds of osteoporosis rats.The changes of bone microstructure parameters could increase bone volume ratio to total volume,trabecular thickness and trabecular number,decrease bone surface area ratio to bone volume and bone trabecular spacing,and increase bone mineral density.In addition,the expression of HSD11B1,CYP19A1,ALOX15 could be down-regulated in low,middle and high dose groups.Conclusion(1)Echinops latifolius has therapeutic effect on both postmenopausal osteoporosis model rats and senile osteoporosis model rats.It could inhibit bone resorption,improve anti-oxidation ability,bone pathological changes,bone microstructure and increase bone mineral density.(2)Phenylpropanoic acids(total phenolic acids)in Echinops latifolius are an important substances for its anti-osteoporosis effect.(3)Total phenolic acid and down-regulation of the expression of HSD11B1,CYP19A1 and ALOX15 in the oxidative reduction pathway may be the most important substances and mechanisms for the treatment of osteoporosis in Echinops latifolius.