Aqueous extract of Protaetia brevitarsis larvae increases mTOR-mediated growth rate in zebrafish larvae

Objective:To evaluate the effects of an aqueous extract of Protaetia brevitarsis(AEPB)on the growth of zebrafish and preosteoblast MC3T3-E1 cells.Methods:The effects of AEPB on the linear growth and the expression of growth-related genes in zebrafish and MC3T3-E1 cells were assessed using various molecular techniques.Furthermore,the involvement of the mammalian target of rapamycin(mTOR)pathway in AEPB-induced growth was investigated by employing the mTOR inhibitor rapamycin.Results:AEPB administration led to a significant and dose-dependent increase in zebrafish larvae growth over time.Additionally,AEPB treatment upregulated the expression of growth hormone-1(GH-1),insulin-like growth factor-1(IGF-1),growth hormone receptor-1(GHR-1),and cholecystokinin-a(CCKA)in zebrafish.Similarly,AEPB stimulated the expression and release of IGF-1 and accelerated mTOR expression in MC3T3-E1 cells.In addition,rapamycin hindered AEPB-induced linear growth in zebrafish larvae and suppressed the expression of growth-promoting genes by inhibiting mTOR activation.Conclusions:AEPB shows growth-promoting effects by upregulating growth-related genes and activating the mTOR signaling pathway.Further investigations are warranted to elucidate its mechanisms of action and explore its potential application in the development of growth-enhancing supplements for various purposes.

Aqueous extract of freeze-dried Protaetia brevitarsis larvae promotes osteogenesis by activating β-catenin signaling

Objective:To investigate the effect of an aqueous extract of Protaetia brevitarsis(AEPB)on osteogenesis using preosteoblast MC3T3-E1 cells and zebrafish larvae.Methods:Flow cytometric analysis was used to measure the cytotoxicy.Alkaline phosphatase activity was detetmined using p-nitrophenyl phosphate as a substrate.Calcium deposition was detected using alizarin red staining along with osteogenic marker expression in preosteoblast MC3T3E1 cells.In addition,vertebral formation in zebrafish larvae was detected using calcein staining and osteogenic gene expression.Results:AEPB highly promoted the expression of osteogenic markers including runt-related transcription factor 2,osterix,and alkaline phosphatase,along with elevated levels of mineralization in MC3T3-E1 cells.Moreover,AEPB accelerated vertebral formation in zebrafish larvae accompanied by upregulated expression of osteogenic genes.FH535,an inhibitor of Wnt/β-catenin,suppressed AEPB-induced osteogenic gene expression and vertebral formation,indicating that AEPB stimulates osteogenesis by activating the Wnt/β-catenin signaling pathway.Conclusions:AEPB stimulates osteoblast differentiation and bone formation by activatingβ-catenin.Therefore,AEPB is a promising material that induces osteogenesis,and is useful for the treatment of bone resorption diseases.