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[Research on osteogenesis metabolism of transgenic zebrafish by hepcidin green fluorescent protein]
1Department of Orthopedics, the Wuxi Second Affiliated Hospital of Nanjing Medical University, Wuxi 214000, China.
Objective:
To investigate the effect of hepcidin downregulation on the osteogenesis metabolism and involved mechanism based on the construction of hepcidin green fluorescent protein (GFP) in the transgenic zebrafish.
Methods:
The eggs of zebrafish were transfected by hepcidin GFP plasmid by microinjection in half an hour after fertilization. The expression of hepcidin in the zebrafish was checked every 6 hours for 120 hours.Iron staining change was done in the larvae of zebrafish transfected by ATG morpholine and GFP plasmid compared with wild type in 4 days in the same conditions.Alizarin red staining of bone and cartilage was performed in 9 days and the expression of bone metabolic cDNA differences was analysed with real-time PCR on 4th day.
Results:
Hepcidin expressed begin on 72 hours and strongest in 96 hours on GFP transgenic zebrafish.Iron staining was significantly darker than the GFP-control type on 4th day. Alizarin red staining became shallow and ribs and caudal fin did not change furthermore in zebrafish transfected by ATG morpholine. PCR related metabolic gene expression was expressed down in zebrafish transfected by ATG morpholine.
Conclusion:
Hepcidin expressed strongest on GFP transgenic zebrafish on 96 hours. The inhibition of hepcidin caused iron accumulation and lowered the osteogenesis led to retardation and reduction of bone in zebrafish.
