The Function Study Of Glucosamine Hydrochloride On Skeletal Development And Repair Using Zebrafish Model

Glucosamine hydrochloride(GAH)is one of the most basic and important derivatives of chitin,which is obtained by hydrolysis of chitin inconcentrated hydrochloric acid.In this research,we use zebrafish model to study the effects and underlying molecular mechanisms of GAH,extracted from the cell walls of Agaricus bisporus,on skeletal development and injury repair in zebrafish larvae and adult.In the first,we determine three concentration gradient of GAH by fatality rate.Our results show that GAH not only promotes cartilage and bone development in larval,but also promotes the caudal fin regeneration in adult,in a dose-dependent manner.We found that GAH treatment significantly cause an increase of bone-related marker genes by Whole-mount in situ hybridization,such as,col2a1a、col10a1a、BGP、sp7、sox9a、sox9b,indicating that GAH plays an important role in promoting bone cartilage development.In addition,we set up an osteoporosis model by treating ferric ammonium citrate(FAC)with zebrafish.And then we determine the function of GAH on this zebrafish FAC-induced osteoporosis model.Our data display that GAH has an ameliorative effect specially on bone cartilage defects,and that GAH treatment can rescue the expression of a series of bone related-gene in juvenile fish and adult fish by performing cartilage and bony staining,in situ hybridization and semi-quantitative PCR analyses.Furthermore,we explore whether TGF β/Bmp signaling is involved in GAH-mediated bone development in zebrafish.Our results show that the components of TGFβ/Bmp(such as bmp2 b and bmp4)can be induced within GAH treatment by Whole-mount in situ hybridization.Meanwhile,GAH not only promotes Bmp2 b expression but also increases the activities of phosphor-Smad1/5/8 in zebrafish embryos by immuno-fluorescence,therefore enhancing Bmp signal transduction.In the last,we compare the effect of GAH and other carbohydrates on zebrafish bone development.Our data show that GAH is better for bone injury repair and prevention than others.Taken together,our results reveal that GAH not only promotes bone growth,but also efficiently benefits the treatment and prevention for bone injury,therefore providing a potential drug for the therapy of bone disease.