OPG基因敲除小鼠骨质疏松情况的研究  

Study on osteoporosis in offspring of OPG knockout mice

作　　者：程少丹[1] 王拥军[1] 唐德志[1] 周泉[1] 李晨光[1] 周重建[1] 施杞[1] 

机构地区：[1]上海中医药大学脊柱病研究所骨代谢疾病研究室

出　　处：《中国骨质疏松杂志》2008年第1期16-19,共4页Chinese Journal of Osteoporosis

基　　金：国家科技部国际合作重点资助项目(2006DFA32670);国家杰出青年科学基金资助项目(30625043);国家教育部"新世纪优秀人才支持计划"资助项目(NCET-05-0418);国家人事部留学回国人员科技活动择优资助项目(20053010);上海市医学领军人才支持计划资助项目(05YLJ018);上海市国际合作重点资助项目(55407070);上海市重点学科建设资助项目(T0303);上海市医学重点学科建设资助项目(05Ⅲ027)

摘　　要：目的研究护骨素(Osteoprotegerin,OPG)基因敲除纯合子小鼠(homozygous OPG knockout mice,OPG-/-mice)骨质疏松发生情况,为OPG-/-小鼠骨质疏松模型的应用提供依据。方法非频密繁殖法获得OPG-/-小鼠,PCR技术进行OPG基因表型鉴定。16周龄OPG-/-子代小鼠和16周OPG野生型小鼠各10只,采用双能X线骨密度测量仪(DEXA)测定全身骨密度(Bone mineral density,BMD)、万能材料试验机测定股骨生物力学强度;骨组织形态计量学分析L5椎体骨小梁结构;实时荧光定量PCR检测L1椎体骨组织中BMP-2、Runx2 mRNA表达水平。结果OPG-/-子代小鼠和亲代纯合子小鼠表现出相同的OPG基因缺失表型。与同龄野生型小鼠比较,16周龄OPG-/-小鼠全身骨密度、股骨承受最大载荷、股骨结构刚度、腰椎椎体骨小梁数目、腰椎椎体骨小梁厚度显著下降(t=5.740,6.069,6.859,6.891,3.558,P<0.01);股骨承受破裂载荷、腰椎椎体骨小梁体积分数下降(t=3.157,3.329,P<0.05);股骨承受载荷后的最大位移、破裂位移显著增加(t=-3.868,-3.276,P<0.01);腰椎椎体骨小梁分离度增加(t=-2.575,P<0.05),腰椎椎体Runx2 mRNA表达升高(t=-3.738,P<0.05);腰椎椎体中BMP-2 mRNA表达升高不明显,差异无统计学意义(t=-1.589,P>0.05)。结论OPG-/-小鼠表现出明显的骨质疏松,是一种理想的骨质疏松模式动物。Objective To study osteoporosis generating in the offspring of homozygous （^-/-） OPG knockout mice. Methods OPG^-1- knockout mice were used to mate. Conditions of parental generation propagating and offspring developing were observed. Gene phenotype was identified with PCR.Total bone mineral density（BMD） were determined using dual energey X-ray absorprion（ DEXA）, femur mechanical properties were evaluated using a three-point test, the sclerous tissues slices of lumbar vertebral body stained with picric acid-fuchsin were used to assess bone tissue morphometry static anylysis,the gene expression of BMP-2 ,Runx2 in the Lj vertebral body were assayed by Real time RT PCR in 10 16-week-old offspring of OPG^-1- mice and 10 16-week-old wild type（ ＋ / ＋ ） offspring of heterozygous（ - / ＋ ） OPG knockout mice. Results OPG^-1 - mice were viable and fertial, the offspring could grow and develop normally and revealed the same OPG deletion with parental generation. Compared with OPG ^＋ /＋ mice, in OPG^-1-mice, Total BMD, femur maximum load, stiffness, lumbar vertebral body Tb. N, Tb. Th decreased obviously（ t = 5.740,6.069,6. 859,6.891, 3.558, P 〈 0.01 ） ; femur break load, lumbar vertebral body Tb. Ar% decreased（ t = 3.157, 3.329, P 〈 0.05） ; femur maximum elong, break elong obviously increased （ t = - 3.868, - 3.276, P 〈 0.01 ） ; lumbar vertebral body Tb. Sp increased（ t = - 2.575, P 〈 0.05） ; lumbar vertebral body gene expression of Runx2 increased（ t = -3.738, P 〈 0.05）; lumbar vertebral body gene expression of BMP-2 increased, but the difference had not statistical significance （t = - 1.589, P 〉 0.05 ）.Conclusions OPG^-1- mice appears severe osteoporosis, and it can serve as an excellent osteoporosis animal model.

关 键 词：骨保护素 基因敲除 小鼠 骨质疏松 动物模型 

分 类 号：R31[医药卫生—基础医学]