Effects Of You Gui Yin On The Pharmacodynamic Evaluation Of Rat Of Steroid-induced Avascular Necrosis Of The Femoral Head And Preliminary Study On The Proteomics

Objective:(1)through repeated bolus injection of endotoxin and methyl prednisolone injection induced the establishment of wistar rats Early SANFH model ideal animal model for the subsequent comparative proteomics research.(2)Providing the behavioral evidence for YouGuiYin prevention and treatment of femoral head necrosis(3)the use of high-throughput LC coupled with identification and screening of a normal control group,model group Youguiyin Treated animals bone tissue proteome comparative analysis to determine the femoral head necrosis in the pathogenesis of the development process of the potential biological markers,and thus to study the treatment of the role of the differentially expressed proteins Youguiyin explore the mechanism of action of Chinese medicine prevention and treatment of femoral head nerosis.Methods:(1)30 male Wistar rats were randomly divided into blank control group,the model group and Youguiyin treatment group.Endotoxin and methyl prednisolone injection induced cause necrosis of the femoral head,the Youguiyin treatment group received 10ml/kg gavage,blank control group and model group to the same amount of normal saline treatment,were killed after a large mice,the animals were observed for general condition,femoral appearance,pathological changes(including fat cells of the bone marrow cavity,trabecular bone,and bone cells)changes.(2)With 30 healthy adult wistar rats cause of femoral head necrosis model using the above method,gavage six weeks with the normal group and model group Youguiyin 10ml/kg grip,spontaneous activity,tail suspension,Kuang field experiments and water maze test behavior observed.(3)Get SANFH Early pathological changes and normal group Yougui drinking intervention group femoral head specimens,the use of high-throughput liquid chromatography-mass spectrometry(HPLC-MS)to create early SANFH with normal femoral head,the Youguiyin group protein Spectrum,at its points of difference will be identified.Results:(1)the model group 6W light microscope visible femoral head necrosis performance,trabecular bone begins to shrink,even thinner fracture the bone karyopyknosis dissolved,disappeared perinuclear empty halo larger increase in the number of empty lacunae osteoblasts decreased bone cell cytosol full of lipid droplets the bone nucleus pushed to the edge,decrease in bone marrow stem cells,or even disappear,the accumulation of a large number of fat cells increase in size,and even fusion,the number increased;the Youguiyin bone marrow cavity the relative number of fat cells is significantly less morphologically normal bone marrow stem cells abundant.(2)model group grip is reduced,reduced autonomic activity,the tail suspension Activity fewer,significant reduction in the total distance of the open field test,lOmin movement.Water maze test,the model group were significantly longer escape latency,reduced total distance of activities.The Youguiyin normal group,the difference was statistically significant behavioral indicators above each finger.(3)To obtain good resolution and repeatability early SANFH and normal femoral head,the You Gui Yin group of protein expression profiles.BSA standard measurements,protein concentration is 0.857 ug/ul model group,with total protein 102.9 ug,protein concentration is 1.407 ug/ul blank group,with total protein 168.8 ug,the right to drink group of protein concentration is 1.444 ug/ul,with total protein 173.3 ug.Identified protein for 1164,a total of 178246 secondary spectra match to the spectra of the number 9487,match to the spectra characteristic of peptides number 8327,identification of peptides to 3959,the number of identification to the number of unique peptides sequence of 3598.Early SANFH Youguiyin treatment group,the protein expression profiles of normal femoral head found blank with the model group,84 proteins were upregulated,67 downregulated,the blank group Youguiyin group,there are 90 kinds of expression raised,48 the the downregulation of,model group Youguiyin the group,of which 58 proteins were up-regulated in the early SANFH,another 34 proteins were down-regulated.In blank group and model group differences in protein,Lgals1,BSP OPN and Mgp,col higher multiples.Conclusions:(1)repeated high-dose injections injected toxins and methyl prednisolone injection method can be induced in Wistar rats the early SANFH model.Youguiyin to improve the role the group SANFH pathology pathology evidence for its prevention and treatment of femoral head necrosis(2)early femoral head necrosis can be more obvious to improve their behavior to learn relevant indicators Youguiyin to Youguiyin prevention of femoral head necrosis related behavioral science evidence of animal experiments.(3)high-throughput LC MS reproducible protein expression profiling.Through the analysis of differentially expressed proteins between identification the early SANFH and normal femoral head and Youguiyin-treatment rich in disease-related differences in proteomics data,preliminary screening out potential You Gui Yin prevention and early disease pathogenesis and SANFH important proteins.(4)In the early days of SANFH,Lgals1,BSP)OPN and Mgp、col higher multiples,that can be used as potential biomarkers of SANFH early,provide theoretical and experimental basis for the further study SANFH pathogenesis and prevention.