Correlation Between Kidney And The Development Of Skeletal System At The Embryonic Stage

Objective:Take advantage of the high specific kidney injury characteristic of cisplatin,to build the model of chicken embryo kidney injury. Analyze the relationship between the development of kidney and the development of skeletal system, use chicken embryo as experimental model to elucidate correlation between kidney and the development of skeletal systemat the embryonic stage.Methods:48 fresh eggs were hatched by thermostatic incubator for 4 days, make small holes at eggshells and eggs were randomly allocated into four groups, one blank control group and three groups treated with cisplatin in different dosage: 25μg/egg, 50μg/egg, 100μg/egg, continue to hatch the eggs and analyze the embryos at the 12 th day. On the fourth and a half day of the development of chicken embryos, embryo limb bud tissues were dissected,and embryo bonecells were cultured in vitro. The bone morphology was observed with HE staining and Alcian blue- Alizarin Red staining.The levels of serum calcium, 25(OH) D, tissue section of alkaline phosphatase(ALP), tartrate-resistant acid phosphatase(TRAP) were measured.Results: The development of kidney has significant differences between the cisplatin treatment group and the PBS control group, the differences are dose-related(P<0.01). The RT-PCR results of Col 2al, ALP, Runx2, SOX9, MMP13, GADPH demonstrate that the expression of genes related to bone were attenuated(p <0.05). The levels of 25(OH) D, tissue sections of alkaline phosphatase(ALP) and tartrate resistant acid phosphatase(TRAP) were decreased graduallywith the increasing dosage of cisplatin(P <0.05).Conclusions:1) The use of cisplatin to establish chicken embryo kidney injury model is feasible, the severity of kidney injury is positivelycorrelated with the dosage;2) Treatment with cisplatin has no influence on the skeletal development at the embryonic stage.3) This projectconfirms that the kidney injury impacts the development of skeletal system at the embryonic stage, mainly impacts the function of osteoblasts and osteoclasts.4)It can provide the theoretical support for the occurrence of congenital bone dysplasia.