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The Model of Skeletal and Cartilage Malformation Induced by the Intraperitoneal Administration of Deoxynivalenol in Mice and the Screening of Relating Distinct Proteins

Author: ZhuXiaoMing
Tutor: YuAiLian;ZhangZhong
School: Taishan Medical College
Course: Pathogen Biology
Keywords: Deoxynivalenol Fusarium enol Mice Skeletal deformities Two-dimensional electrophoresis
CLC: R682
Type: Master's thesis
Year: 2010
Downloads: 25
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Abstract


Objective To establish deoxynivalenol sickle bacteria caused enol (deoxynivalenol, DON) the mouse skeletal deformities model; application of two-dimensional electrophoresis screening due to deformed bones differences protein normal fetal rat bones and DON To further elucidate the molecular DON induced skeletal deformities The mechanism of the foundation. Method 1, intraperitoneal injection of four doses of DON exposure, to obtain fetal rat bone, bone cartilage double staining staining dissecting microscope to observe changes in the fetal rat bone cartilage morphology. 2, application grinding method, ultrasonic lysis method and the lysate was extracted total protein of bone and cartilage in the fetal rat vertebrae. Bradford protein concentration determination kit for determination of protein concentration, was extracted to determine the sample volume. 3, using Ettan IPGphor Isoelectric Focusing System Hoefer SE600 separation of total protein silver staining, to obtain two-dimensional electrophoresis gel maps were analyzed using Image Master software normal the control group 10mg/kgDON processing histone point quantitative changes, screening differentially proteins. Results 1, DON affect toxin treatment group the average weight of the fetal rat fetal weight and litter size were 4 mg / kg group 1.10 ± 0.09 g, 6.4 mg / kg group, 0.98 ± 0.08g, 8.4mg/kg group 0.90 ± 0.10 g and 10 mg / kg group, 0.81 ± 0.09 g. Toxin group, the average litter size of 4 mg / kg group 6.83 ± 3.87, 6.4 mg / kg group 6.00 ± 3.63 and 5.00 ± 3.29 8.4 mg / kg group and 10 mg / kg group 5.17 ± 2.71 rats . Experimental group fetal weight and litter size blank control group, the differences were statistically significant, P lt; 0.05, DON experiments on the effect of fetal rat bone cartilage there were more than 35 kinds of bone cartilage defects main axial skeleton malformations, limb bones cartilage deformity followed by multiple skeletal deformities common. Which the neck total of nine kinds of bone cartilage deformity, cervical vertebra fusion, cervical body missing, cervical arch fusion, cervical rib.. The sternal ribs there were six kinds of malformations, including sternum missing, missing ribs, ribs and sternum asymmetric fusion, fusion ribs, both sides of the ribs and the number of different forked ribs, six kinds of the parts of malformations are more common. Thoracic lumbar total of 15 kinds of deformities, missing thoracic vertebrae, lumbar spondylolysis fusion the thoracic vertebrae fragmentation main. The experiment also malformed limbs, two kinds of tail deformities and a variety of other parts of the deformity. Four doses of 10 mg / kg group deformity caused most species, the most severe deformity. The 10mg/kg group cervical body fusion neck deformity deformity rate of cervical missing malformation rate of cervical arch fusion malformation rate, cervical rib malformations were higher than the control group, the difference was statistically significant, P lt; 0.05. The the sternal rib deformity in 10 mg / kg group sternum missing malformation rate, the ribs missing deformity rate fusion rib deformity rate, the number of both sides of the ribs of varying malformation rate, forked rib deformity rate asymmetry sternum connection malformation rate higher than the two control group, the difference was statistically significant, P lt; 0.05. Thoracic vertebrae, thoracic lumbar deformity of 10 mg / kg group missing malformation rate of fusion and lumbar vertebral arch deformity rates were higher than the control group, the difference was statistically significant, P lt; 0.05. In addition 10 mg / kg group the six toe deformity rate capuchin deformity rate is higher than the other two groups, the difference was statistically significant, P lt; 0.05 the deformed bone cartilage protein caused by two-dimensional electrophoresis of DON experimental results were successfully extracted the total protein of fetal rat spinal bone cartilage to obtain two-dimensional electrophoresis map of 10mg/kgDON induced mice bone cartilage deformity, 22 protein spots were screened by the Image Master software analysis, which raised three spots down the protein spots 19. Conclusion 1, DON lower in pregnant rats per litter size and fetal weight with embryo toxicity; 2, DON induced mouse skeletal deformities role, and induced a variety of bone cartilage deformity, axial skeleton malformations based. The intraperitoneal injection 10mg/kgDON lead the multiple bone cartilage deformities, can be used for the preparation of fetal rat skeletal deformities model; 3, the application of two-dimensional electrophoresis technology DON induced mouse the deformed bone cartilage protein electrophoresis gel maps, screened 22 protein spots, three raised spots, down 19 spots, and lay the foundation for further study the mechanisms of DON cause skeletal deformities.

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CLC: > Medicine, health > Surgery > Orthopaedic Surgery ( movement system diseases,orthopedic surgery ) > Bone deformities
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