改良型激素性股骨头坏死小鼠模型的建立与评价Est ablishment and Evaluation of a Modified Mouse Model of Steroid Induced Osteonecrosis of the Femoral Head
王继涛,阮红峰,付方达,吴承亮
摘要(Abstract)：
目的为建立有效的动物模型,更好研究激素性股骨头坏死的发病机制,科学阐明中药方剂治疗该疾病的分子作用机制。方法取45只2月龄雄性C57BL/6小鼠随机平均分为对照组、传统组、改良组。传统组给予脂多糖联合甲强龙激素造模,改良组小鼠则在传统组基础上调整饲料槽的高度,迫使小鼠主动站立取食,并予跑步机上跑步2 h/d;对照组给予等量的生理盐水。定时观察小鼠一般状况,分别于造模前、造模后第4周、第8周抽血检测血液中三酰甘油的含量,并于造模后8周测量小鼠股骨头骨密度,次日处死取双侧股骨头,体视显微镜下观察股骨头外观及色泽。采用苏木精-伊红染色法(HE)及阿尔新蓝染色(ABH)等技术观察小鼠股骨头病理变化。结果传统组和改良组小鼠精神状态明显差于对照组,且改良组差于传统组(P<0.05);传统组和改良组血液中三酰甘油的含量高于对照组,改良组高于传统组;骨密度检测显示,传统组和改良组骨密度低于对照组,且改良组低于传统组(P<0.05);股骨头形态学观察发现改良组股骨头软骨下出血坏死的面积较传统组严重,HE、ABH结果显示,传统组有9只出现(空骨陷窝数量增多,骨小梁变细甚至断裂)等骨坏死表现,造模成功率达60%,改良组有11只出现(空骨陷窝数量增多,骨小梁变细甚至断裂)股骨头坏死的表现,造模成功率高达73.3%(P<0.05)。结论传统造模方法和改良造模方法皆可造模成功,但改良造模方法具有更高成模率。
关键词(KeyWords)： 激素性股骨头坏死;动物模型;甲强龙;脂多糖;跑步机
基金项目(Foundation): 国家自然科学基金项目(81273770,8157150451);; 浙江省自然科学基金项目(LY15H270012,Q17H270023);; 浙江省中医药科技计划项目(2017ZB026)
作者(Author): 王继涛,阮红峰,付方达,吴承亮
参考文献(References)：
- [1]Fan SH,Wang YY,Lu J,et al.CERS2 suppresses tumor cell invasion and is associated with decreased V-ATPase and MMP-2/MMP-9 activities in breast cancer[J].J Cell Biochem,2015,116(4):502-513.
- [2]Xie M,Hu A,Luo Y,et al.Interleukin-4 and melatonin ameliorate high glucose and interleukin-1βstimulated inflammatory reaction in human retinal endothelial cells and retinal pigment epithelial cells[J].Mol Vis,2014(20):921-928.
- [3]Lieberman JR,Engstrom SM,Meneghini RM,et al.Which factors influence preservation of the osteonecrotic femoral head[J].Clin Orthop Relat Res,2012,470(2):525-534.
- [4]李子荣.股骨头坏死:早期诊断与个体化治疗[J].中国矫形外科杂志,2013,21(19):1909-1911.
- [5]Tang C,Chen L,Gu W,et al.Cyclosporin A enhances the ability of trophoblasts to displace the activated human umbilical vein endothelial cell monolayers[J].Int J Clin Exp Pathol,2013,6(11):2441-2450.
- [6]Ryoo S,Lee S,Jo S,et al.Effect of lipopolysaccharide(LPS)on mouse model of steroid-induced avascular necrosis in the femoral head(ANFH)[J].J Microbiol Biotechnol,2014,24(3):394-400.
- [7]王坤正,王春生,武永刚,等.激素性骨坏死血管变化的实验研究[J].中华医学杂志,2006,92(29):2024-2027.
- [8]Saito S,Ohzono K,Ono K.Early arteriopathy and postulated pathogenesis of osteonecrosis of the femoral head.The intracapital arterioles[J].Clin Orthop Relat Res,1992(277):98-110.
- [9]边焱焱.大剂量糖皮质激素对骨髓间充质干细胞成骨分化的Miro RNA调节的实验研究[D].北京:清华大学医学部,2012.
- [10]张国桥,易诚青,滕松松,等.早期激素性股骨头坏死模型的建立[J].现代生物医学进展,2012,12(7):1223-1224,1235.
- [11]Bejar J,Peled E,Boss JH.Vasculature deprivation-induced osteonecrosis of the rat femoral head as a model for therapeutic trials[J].Theor Biol Med Model,2005(2):24.
- [12]Bekler H,Uygur AM,Gke A,et al.The effect of steroid use on the pathogenesis of avascular necrosis of the femoral head:an animal model[J].Acta Orthop Traumatol Turc,2007,41(1):58-63.
- [13]Okazaki S,Nishitani Y,Nagoya S,et al.Femoral head osteonecrosis can be caused by disruption of the systemic immune response via the toll-like receptor 4 signalling pathway[J].Rheumatology(Oxford),2009,48(3):227-232.
- [14]黄思俊,白志强,张弛,等.采用激素联合内毒素建立兔中期股骨头坏死模型[J].中华关节外科杂志:电子版,2016,10(3):331-336.
- [15]朱嘉,方以群,金晶,等.激素性股骨头坏死动物模型建立方法初探[J].海军医学杂志,2009,30(2):97-101.