摘要
目的：从成骨细胞（osteoblast，0B）水平探讨核网子κB受体活化子配体（receptor activator of NFκB ligand.RANKL）、骨保护蛋白（osteoprotegerin，OPG）与青少年特发性脊柱侧凸（adolescent idiopathic scoliosis，AIS）患者骨量降低的关系。方法：AIS患者（AIS组）20例，男5例，女15例，年龄11-19岁，平均14.75岁，Cobb角40°-96°，平均59.65°；同年龄非脊柱畸形患者（对照组）8例，男6例，女2例，年龄10-19岁，平均15.25岁。两组均采用双能x线吸收测量仪（dual-energy X-ray absorptiometry，DEXA）测量骨密度（bone mineral density，BMD），测量部位包括非优势侧股骨近端及腰椎。所有受试者术中取适量髂前上嵴的松质骨，运用植块法培养成骨细胞。培养过程巾观察细胞形态学变化，并用碱性磷酸酶染色法、钙结节染色法和RT-PCR检测骨钙素表达，并进行表型鉴定。取P2代细胞应用RT-PCR和Western blotting检测RANKL和OPG的mRNA及蛋白表达情况。结果：植块法培养可以获得较多的原代细胞。碱性磷酸酶染色、钙结节染色及RT-PCR检测骨钙素表达证实所培养的细胞表现成骨细胞特性。AIS组腰椎（L2-L4）及股骨近端的BMD值明显低于对照组；RANKL的核酸及蛋白水平的表达量均较对照组岛（P〈0.01）；OPG的核酸及蛋白水平的表达量与对照组比较无统计学筹异（P〉0.05）；RANKL／OPG比值明显高于对照组（P〈0.01）。结论：成骨细胞中RANKL及OPG在核酸及蛋白水平的表达异常可能与AIS患者骨量降低的分子机制相关。
Objective：To investigate the relationship between receptor activator of nuclear factor κB ligand （RANKL） or osteoprotegerin（OPG） and the decreased bone mass of the patients with adolescent idiopathic seoliosis at the osteoblast level.Method：This study covered 28 subjects （aged from 11 to 19 years）,including patients with AIS and age-matched patients without AIS who underwent spinal surgery.AIS group was 20 patients,control group was 8 cases.The average Cobb angle of AIS group was 59.65°（range,from 40°to 96°）. Bone mineral density （BMD） of the lumbar spine and proximal femur was measured by dual energy X-ray absorptiometry （DEXA） for both groups.The cancellous bone was harvested from the posterior superior iliac spine of each subject,which then were cultured to obtain the osteoblasts by explant culture method.The mor- phology of the cells and formation of calcium nodules were observed,the osteocalcin was determined by staining method of alkaline phosphatase （AKP） and calcium nodules,and RT-PCR.The cell phenotype was then confirmed.Expression of mRNA and proteins of RANKL and OPG was detected by RT-PCR and Western blotting using P2 generation osteoblasts from two groups.Result：More primary cells could be obtained by the explant culture.The expressed osteocalcin dertmined by staining of AKP and calcium nodules and RT-PCR confirmed that phenotype of the cultured cells showed osteoblastic property.BMD of the lumbar spine （L2-IA）and proximal femur was obviously lower in AIS group than that in the control group.RT-PCR and Western blotting results showed that expression of RANKL of osteoblasts increased obviously in AIS group as compared with the control group （P〈0.01）.However, expression of OPG did not have any significant difference between two groups （P〉0.05）.The ratio of RANKL and OPG in patients with AIS was hihger than that in control subjects （P〈0.01）.Conclusion：The imbalance and the disturbed interaction of RANKL and OPG in osteoblasts at nucleic acid
出处
《中国脊柱脊髓杂志》
CAS
CSCD
北大核心
2009年第3期216-221,I0001,共7页
Chinese Journal of Spine and Spinal Cord
基金
江苏省卫生厅重点项目(项目编号:K200610)
