强直性脊柱炎滑膜细胞破骨表型及分化因素的研究  

Osteoclast phenotypes and differentiation factors of synoviocyte in ankylosing spondylitis

作　　者：钱奕铭[1] 初同伟[1] 周跃[1] 张峡[1] 刘玉刚[1] 李建明[1] 

机构地区：[1]第三军医大学新桥医院骨科,重庆400037

出　　处：《第三军医大学学报》2008年第23期2207-2210,共4页Journal of Third Military Medical University

摘　　要：目的通过检测破骨细胞(osteoclast,OC)在强直性脊柱炎(ankylosing spondylitis,AS)骶髂关节及髋关节滑膜组织中的表达及分化因素的研究,探讨AS骨与软骨破坏机制。方法酶组织化学染色技术(TRAP染色)检测滑膜细胞破骨表型,原位杂交技术检测滑膜组织中TNF-α mRNA、VEGF mRNA、MMP-3 mRNA的表达;培养正常滑膜成纤维细胞,加入TNF-α、VEGF,培养AS滑膜成纤维细胞,加入抗TNF-α、抗VEGF,钙钴法检测碱性磷酸酶(ALP)表达,放射免疫法检测骨钙素(bone gla protein,BGP)含量,分析其在AS滑膜成纤维细胞成骨中的作用。结果经TRAP染色,TNF-α mRNA、VEGF mRNA和MMP-3 mRNA在AS滑膜组织中阳性表达,而对照组阴性表达,两组比较有统计学差异(P<0.01);AS组滑膜TRAP染色阳性细胞同时表达MMP-3;AS组滑膜成纤维细胞培养ALP染色阳性数、BGP分泌增加值与正常对照组比较有统计学差异(P<0.01);TNF-α、VEGF单克隆抗体可以抑制滑膜成纤维细胞骨钙素的分泌。结论AS滑膜组织中表达TNF-α、VEGF,两种因子可以诱导滑膜成纤维细胞分化为成骨细胞(osteoblast,OB),在这种OB存在的条件下,滑膜组织内破骨细胞前体可以分化为成熟的OC并表达MMP-3。Objective To investigate whether there exists osteoclasts and differentiation factors in the synovial tissue of patients with ankylosing spondylitis （AS） for the mechanism of destruction of bone and cartilage in AS. Methods Enzymohistochemical staining （TRAP staining） was used to detect the expression of osteo-clast phenotype, and hybridization in situ to detect the mRNA expressions of TNF-α, VEGF and MMP-3 in synovial membrane samples of 21 AS patients and 21 fracture subjects. Synovial cells isolated from normal group were cultured in presence of TNF-α and VEGF, while those from AS group were cultivated with anti-TNF-α and anti-VEGF antibodies. Gomori method was used to detect alkaline phosphatase （ALP） and radioimmunoassay to detect bone gla protein （BGP） in these cultured cells for the role of differential factors in osteoblast phenotype expression. Results The synovial membrane in AS were positive to mRNA expressions of TNF-α, VEGF, and MMP-3, and to TRAP staining, but that of control group were negative. Significant difference was seen in the positive cell numbers between these 2 groups （P 〈0.01 ）. In cultivated synovioblast from AS group, ALP staining was positive, and BGP content was elevated compared with control group. Anti-VEGF and anti-TNF-α inhibited synovioblast to secrete BGP. Conclusion An obvious increase of TRAP positive cells in synovium may provide a main source of osteoclastogenesis in AS patients. With induction of TNF-α and VEGF, synovioblast may differentiate into osteoblasts, and express ALP and BGP. In this condition, synovial mononuclear macrophage may differentiate into osteoclasts and express MMP-3, which play important role in bone resorption.

关 键 词：强直性脊柱炎 滑膜细胞 破骨细胞 原位杂交技术 细胞因子 TRAP染色 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] R392.13[医药卫生—基础医学]