Early-stage Proteomics Of Steroid-induced Avascular Necrosis Of Femoral Head

There is an increasing tendency in the incidence of SANFH(steroid-induced avascular necrosis of femoral head). However,The pathogenesis of SANFH has not been totally understood until now.Every function in the living cell depends on proteins, proteins play an important role in the occurrence of disease, development, and implementation of the vesting. Post-genomic era as a prelude to the opening up of the climax of proteomics. Although in its infancy, Proteomics has been widely used in orthopedic diseases, which can be applied to a new SANFH research methods and a new model. And it has not yet been reported about the dynamic study on the relationship of protein molecules and intake of steroid leading to SANFH.First, we establish a stable animal model for steroid-induced avascular necrosis of femoral head in rabbits :①"LPS+MP";②"MP";③"HS+MP". The goal of this study is to construct a dynamic comparative proteome database of SANFH and normal femoral head tissues in different time points. The obtained results will provide a theoretical and experimental basis for further exploration on the SANFH.Part 1 the animal model Constructions of steroid-induced avascular necrosis of femoral headTo establish a stable animal model for steroid-induced avascular necrosis of femoral head in rabbits. The obtained results will provide a theoretical and experimental basis for further exploration on the SANFH.Fourty-four adult New Zealand rabbits were randomly divided into four groups: (1)group A :"LPS+MPS"(n=12) (;2)group B :"MPS"(n=12) ; (3)group C :"HS+MPS"(n=12) ; (4)group D :"NS+NS"(n=8) .。The percentage of trabecular bone area and empty lacunae and microvascular density were measured。methylprednisolone.Histopathological changes in the femoral heads were observed by light microscope and transmission electron microscope at the end of six weeks after the injection Methylprednisolone combined with lipopolysaccharide can induce typical rabbit model for early avascular necrosis of femoral head.Part 2 early-stage Proteome study of steroid-induced avascular necrosis of femoral headTo establish a stable protein extraction method for steroid-induced avascular necrosis of femoral head in rabbits.For achieving a high quality on resolution and a steady result, effort was made to establish and optimize the 2-DE technique that should be adaptive for proteomic analysis on bone tissue. Establishment of this technical platform provided the foundation for following research.For achieving a high quality on resolution and a steady result, effort was made to establish and optimize the 2-DE technique that should be adaptive for proteomic analysis on bone tissue samples. The conventional protocol of protein extraction was modified, especially on some critical steps such as sample processing (grinding,dialysising and acetone precipitation), quantitative methods of protein (Lowery method and modified Bradford method), loading modes (cup-loading and swelling-loading), IEF procedure, and etc.The protein spots from same sample that run repeatedly three times of 2-DE were 423 and 468 respectively. Establishment of this technical platform provided the foundation for following research. Conclusion1. Methylprednisolone combined with lipopolysaccharide can induce typical rabbit model for early avascular necrosis of femoral head.2. acetone precipitation of proteins can be used in this study .3. The modified 2-DE technical platform for proteomic analysis on bone tissues provides the stable foundation for research.