Construction method for zebra fish model using hepcidin to treat high-iron inhibition of bone formation

Abstract

The invention discloses a construction method for a zebra fish model using hepcidin to treat high-iron inhibition of bone formation. The method comprises the following steps: with zebra fish oosperms as a model, carrying out intervention by using ferric ammonium citrate (FAC) of different concentrations, carrying out alizarin red staining and calculating bone mineralization areas at the 4th day and selecting the optimal intervention concentration of FAC; choosing zebra fish oosperms which are in cell stage 1 to 2 under a microscope, injecting hepcidin mRNA with Flag labels and of different concentrations, placing injected oosperms in an FAC culture solution with a concentration of 100 mu g / mL, carrying out intervention and then carrying out alizarin red staining at the 4th day so as to obtain the zebra fish model using hepcidin to treat high-iron inhibition of bone formation. In recent years, considerable research shows that iron metabolism is related to bone metabolism; a zebra fish is advantageous as a model since the zebra fish develops osteoclasts in 20 d after fertilization, and the zebra fish model can be used for effective research on influence of iron metabolism on bone formation. Meanwhile, commercially available hepcidin has the disadvantages of a high price, few manufacturers and uncertain biological activity at present; hepcidin used in the model is originated from a zebra fish sequence and has certain biological activity.

Description

technical field [0001] The invention relates to the field of medical technology, in particular to a method for constructing an animal experiment model, in particular to a method for constructing a zebrafish model in which hepcidin treats high iron to inhibit bone formation. Background technique [0002] Iron ions are essential trace elements for the human body and have important biological functions. "Iron overload" occurs when iron ions in the blood exceed the binding capacity of transferrin to form free iron that is not bound by transferrin. Clinically, in order to facilitate the study of the correlation between iron overload and disease, serum ferritin greater than 1000 μg / L is defined as "iron overload". Iron overload is common in conditions such as hemochromatosis, thalassemia, and sickle cell anemia. In recent years, studies have found that iron accumulation exists in some postmenopausal women, that is, the serum ferritin level is greater than the normal high value o...

AI Technical Summary

Problems solved by technology

However, currently commercially available hepcidin is only produced by a Japanese bio-company. It is expensive, has controversial biological activity, and is not suitable for storage. Once opened, it is very easy to deteriorate. Therefore, this commercially available hepcidin cannot be widely used in basic and clinical research.

[0004] There is no ideal zebrafish model in which hepcidin treatment of high iron inhibits bone formation

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