Objective:In traditional Chinese medicine,the herbal pair,Radix Achyranthis Bidentatae (RAB) and Eucommiae Cortex (EC),is widely used to treat osteoporosis.Herein,we determined whether this herbal pair can be used to ameliorate glucocorticoid (GC)-induced osteoporosis (GIOP) and find its optimal dosage in zebrafish.Methods:The characteristics of the aqueous extract of RAB and EC were separately characterized using high-performance liquid chromatography.Osteoporosis was induced in 5-day post-fertilization zebrafish larvae by exposing them to 10 μmol/L dexamethasone (Dex) for 96 h.Seven combinations of different ratios of RAB and EC were co-administered.Treatment efficacy was determined by calculating zebrafish vertebral area and sum brightness,via alizarin red staining,and by detecting alkaline phosphatase (ALP)activity.Multiple regression analysis was conducted to test the optimal dosage ratio.Results:According to the Chinese Pharmacopoeia (2015),β-ecdysone (β-Ecd) is a major bioactive marker in RAB extract,while pinoresinol diglucoside (PDG) is the major marker in EC extract.Both of β-Ecd and PDG content values aligned with the Chinese Pharmacopoeia standards.Treatment with 10 μmol/L Dex reduced zebrafish vertebral area,sum brightness,and ALP activity,but RAB and EC attenuated these effects.Combining 50 μg/mL RAB and 50 μg/mL EC was optimal for preventing GIOP in zebrafish.Reverse transcription-quantitative polymerase chain reaction was used to evaluate the mRNA expression of osteogenesis-related genes.A treatment of 10 μmol/L Dex decreased runt-related transcription factor 2(Runx2),osteogenic protein-1 (OP-1),bone γ-carboxyglutamic acid-containing protein (BGLAP),andβ-catenin levels.This effect was counteracted by RAB and EC co-treatment (P ＜ 0.05).Additionally,the effect of using the two herbal extracts together was better than single-herb treatments separately.These results demonstrated that RAB and EC preserve osteoblast function in the presence of GC.The best mass ratio was 1∶1.Conclusion:RAB and EC herbal pair could ameliorate GC-induced effects in zebrafish,with 1∶1 as the opti-mal dosage ratio.

Joon-ho Lee;Yuan-ji Wei;Zhong-yan Zhou;Yu-ming Hou;Cheng-long Wang;Li-bo Wang;Hong-jin Wu;Yu Zhang;Wei-wei Dai

Central Laboratory of Science and Technology Department,Longhua Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China;Cardiovascular Department,Longhua Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China

结合医学学报（英文版）

[1]Joon-ho Lee;Yuan-ji Wei;Zhong-yan Zhou;Yu-ming Hou;Cheng-long Wang;Li-bo Wang;Hong-jin Wu;Yu Zhang;Wei-wei Dai-.Efficacy of the herbal pair,Radix Achyranthis Bidentatae and Eucommiae Cortex,in preventing glucocorticoid-induced osteoporosis in the zebrafish model)[J].结合医学学报（英文版）,2022(01):83-90

Achyranthis,Bidentatae,Eucommiae

Efficacy,herbal,pair,Radix,Cortex,preventing,glucocorticoid,induced,osteoporosis,zebrafish,model,Objective,In,traditional,Chinese,medicine,RAB,EC,widely,used,Herein,determined,whether,can,ameliorate,GIOP,find,its,optimal,dosage,Methods,characteristics,aqueous,were,separately,characterized,using,high,performance,liquid,chromatography,Osteoporosis,was,day,post,fertilization,larvae,by,exposing,them,dexamethasone,Dex,Seven,combinations,different,ratios,administered,Treatment,efficacy,calculating,vertebral,area,sum,brightness,via,alizarin,staining,detecting,alkaline,phosphatase,ALP,activity,Multiple,regression,analysis,conducted,test,Results,According,Pharmacopoeia,ecdysone,Ecd,major,bioactive,marker,while,pinoresinol,diglucoside,PDG,Both,content,values,aligned,standards,reduced,but,attenuated,these,effects,Combining,Reverse,transcription,quantitative,polymerase,chain,reaction,evaluate,expression,osteogenesis,related,decreased,runt,Runx2,osteogenic,protein,bone,carboxyglutamic,acid,containing,BGLAP,catenin,levels,This,counteracted,Additionally,two,extracts,together,better,than,single,treatments,These,results,demonstrated,that,preserve,osteoblast,function,presence,best,mass,Conclusion,could

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