脂联素对小鼠股骨骨折修复的促进作用及其机制探索
发布时间：2018-11-26 17:54
【摘要】：目的：建立小鼠股骨线性骨折模型,观察小鼠骨折愈合过程观察脂联素对小鼠股骨骨折修复的促进作用,初步探讨脂联素对小鼠股骨骨折修复的机制。方法：研究对象选取C57BL/6J小鼠,建立小鼠股骨骨折模型,根据药品、用药时间及用药剂量的不同分为12个组。观察各组小鼠松质骨的体积及皮质骨的厚度,生物力学强度及骨折断端愈合情况的变化。应用EL ISA检测血指标,利用Western Blotting表达分析骨折部位BMP-2的变化,同时观察Masson三色染色中骨性骨痂的形成过程。结果：1.骨折部位大体观察、HE染色及Micro-CT三维重建图显示：脂联素组及BMP-2组2周骨折线已开始模糊,4周骨折线已几乎不可见,6周塑形基本完成,优于对照组的骨折修复；2.骨折部位生物力学检测显示：最大载荷、最大桡度、弹性桡度、刚度这四个指标在脂联素组及BMP-2组均高于阴性对照组(p0.05);3. Micro-CT三维兴趣区与指标BMD、BV/TV结果显示：与对照组相比,脂联素组及BMP-2组的骨皮质连续性及骨松质结构优于阴性对照组,BMD及BV/TV在脂联素用药组及BMP-2组的水平也高于对照组(p0.05)。4.血检测指标显示：BMP-2、脂联素、OCN、IL-1、IL-6、IL-17、ROS检测差异无统计学意义(p0.05)。OPG在手术2周及4周后各组的表达水平随时间的推移而升高,在术后4周时脂联素2mg/kg组的OPG水平高于其他各组,OPG的表达在6周时降低(p0.05)。5Western Blotting表达分析：手术2周后,BMP-2组的BMP-2表达量高于其他各组,手术4周和6周后脂联素2mg/kg组的BMP-2表达量高于其他各组;6Masson三色染色：脂联素组及BMP-2组在2周时骨性骨痴明显多于阴性对照组,4周时脂联素组及BMP-2组骨痂明显增多且已开始塑形,6周时用药组骨痂改建基本完成。结论：脂联素可明显增加小鼠骨折断端的骨痴形成和骨皮质连续性;提高骨折修复后股骨的强度、弹性：改善了骨折部位骨皮质及骨松质的骨微结构。局部BMP-2的表达增加,可能是脂联素局部作用促进骨折修复的机制之一。
[Abstract]:Objective: to establish a linear fracture model of femur in mice and observe the promoting effect of adiponectin on the repair of femur fracture in mice during the course of fracture healing, and to explore the mechanism of adiponectin in repairing fracture of femur in mice. Methods: C57BL/6J mice were selected to establish femur fracture model and divided into 12 groups according to drug, time and dosage. The volume of cancellous bone, the thickness of cortical bone, the biomechanical strength and the healing of fracture were observed. EL ISA was used to detect blood indexes, Western Blotting expression was used to analyze the changes of BMP-2 in fracture site, and the formation process of bone callus in Masson trichrome staining was observed. Results: 1. Gross observation of fracture site, HE staining and Micro-CT 3D reconstruction showed that the fracture line of adiponectin group and BMP-2 group had begun to blur at 2 weeks, the fracture line at 4 weeks was almost invisible, and the plastic shape was basically completed at 6 weeks, which was superior to the fracture repair in control group. 2. Biomechanical examination of fracture site showed that the maximum load, maximum radial, elastic radial and stiffness were higher in adiponectin group and BMP-2 group than in negative control group (p0.05). Compared with the control group, the continuity and cancellous structure of bone cortex in adiponectin group and BMP-2 group were superior to those in negative control group. The levels of BMD and BV/TV in adiponectin group and BMP-2 group were also higher than those in control group (p0.05). There was no significant difference in adiponectin (BMP-2,) and OCN,IL-1,IL-6,IL-17,ROS (p0. 05). OPG expression level increased with time after 2 and 4 weeks of operation. The level of OPG in adiponectin 2mg/kg group was higher than that in other groups at 4 weeks after operation, and the expression of OPG decreased at 6 weeks (p0.05). Analysis of 5Western Blotting expression: after 2 weeks of operation, the expression of BMP-2 in BMP-2 group was higher than that in other groups. The expression of BMP-2 in adiponectin 2mg/kg group was higher than that in other groups after 4 and 6 weeks of operation. 6Masson trichromatic staining: the osteosclast in adiponectin group and BMP-2 group was significantly higher than that in negative control group at 2 weeks, the callus in adiponectin group and BMP-2 group increased significantly at 4 weeks, and the callus remodeling was basically completed at 6 weeks. Conclusion: adiponectin can significantly increase the formation of bone eccentricity and the continuity of bone cortex, increase the strength and elasticity of femur after fracture repair, and improve the bone microstructure of bone cortex and cancellous substance in fracture site. The increased expression of local BMP-2 may be one of the mechanisms of adiponectin in promoting fracture repair.
【学位授予单位】：中国人民解放军医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R683.4
本文编号：2359229
[Abstract]:Objective: to establish a linear fracture model of femur in mice and observe the promoting effect of adiponectin on the repair of femur fracture in mice during the course of fracture healing, and to explore the mechanism of adiponectin in repairing fracture of femur in mice. Methods: C57BL/6J mice were selected to establish femur fracture model and divided into 12 groups according to drug, time and dosage. The volume of cancellous bone, the thickness of cortical bone, the biomechanical strength and the healing of fracture were observed. EL ISA was used to detect blood indexes, Western Blotting expression was used to analyze the changes of BMP-2 in fracture site, and the formation process of bone callus in Masson trichrome staining was observed. Results: 1. Gross observation of fracture site, HE staining and Micro-CT 3D reconstruction showed that the fracture line of adiponectin group and BMP-2 group had begun to blur at 2 weeks, the fracture line at 4 weeks was almost invisible, and the plastic shape was basically completed at 6 weeks, which was superior to the fracture repair in control group. 2. Biomechanical examination of fracture site showed that the maximum load, maximum radial, elastic radial and stiffness were higher in adiponectin group and BMP-2 group than in negative control group (p0.05). Compared with the control group, the continuity and cancellous structure of bone cortex in adiponectin group and BMP-2 group were superior to those in negative control group. The levels of BMD and BV/TV in adiponectin group and BMP-2 group were also higher than those in control group (p0.05). There was no significant difference in adiponectin (BMP-2,) and OCN,IL-1,IL-6,IL-17,ROS (p0. 05). OPG expression level increased with time after 2 and 4 weeks of operation. The level of OPG in adiponectin 2mg/kg group was higher than that in other groups at 4 weeks after operation, and the expression of OPG decreased at 6 weeks (p0.05). Analysis of 5Western Blotting expression: after 2 weeks of operation, the expression of BMP-2 in BMP-2 group was higher than that in other groups. The expression of BMP-2 in adiponectin 2mg/kg group was higher than that in other groups after 4 and 6 weeks of operation. 6Masson trichromatic staining: the osteosclast in adiponectin group and BMP-2 group was significantly higher than that in negative control group at 2 weeks, the callus in adiponectin group and BMP-2 group increased significantly at 4 weeks, and the callus remodeling was basically completed at 6 weeks. Conclusion: adiponectin can significantly increase the formation of bone eccentricity and the continuity of bone cortex, increase the strength and elasticity of femur after fracture repair, and improve the bone microstructure of bone cortex and cancellous substance in fracture site. The increased expression of local BMP-2 may be one of the mechanisms of adiponectin in promoting fracture repair.
【学位授予单位】：中国人民解放军医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R683.4
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