摘要
目的:建立关节炎大鼠滑膜成纤维细胞和单核细胞共培养诱生破骨样细胞的方法,研究清络通痹颗粒对破骨样细胞分化、增殖的影响及机制。方法:分离佐剂性关节炎大鼠外周血单核细胞和滑膜组织,将滑膜成纤维细胞和单核细胞共培养诱生破骨样细胞并以TRAP染色鉴定;制备清络通痹颗粒含药血清,加入共培养体系,MTT法检测破骨样细胞增殖、TRAP试剂盒检测培养上清TRAP活性、扫描电镜观察骨吸收作用、ELISA法检测培养上清中IL-1、TNFα-和RANKL水平。结果:关节炎大鼠滑膜成纤维细胞和外周血单核细胞共培养可诱生出破骨样细胞;清络通痹颗粒3.6、7.2、14.4 g/kg给药的大鼠含药血清可不同程度抑制破骨样细胞的增殖、TRAP活性和骨吸收,降低培养上清中IL-1、TNFα-和RANKL的水平。结论:清络通痹颗粒可抑制共培养诱生的破骨细胞的分化和活性,这一作用与抑制共培养体系中细胞因子的过度分泌有关。
Objective：To set up the method for inducing osteo-like cells by co-culturing synovial fibroblasts and monocytes of arthritic rats and investigate the effects of Qingluotongbi granule（QLT） on the differentiation and activity of the osteo-like cells.Methods：The monocytes of peripheral blood and synovial tissue of adjuvant-induced arthritic rats were separated,and synovial fibroblasts and monocytes were co-cultured to induce osteo-like cells.The cells were identified by TRAP staining.QLT-containing sera were obtained and added to the co-cultured system,and the proliferation was detected by MTT assay,TRAP activity was detected by assay kit,bone resorption area of osteo-like cells was detected by scanning electron microscope and the levels of IL-1,TNF-α and RANKL were detected by ELISA.Results：Co-culturing synovial fibroblasts and monocytes could induce osteo-like cells.QLT-containing sera could inhibit the proliferation,TRAP activity and bone resorption area of osteo-like cells,and the sera could also reduce the levels of IL-1,TNF-α and RANKL in the supernatants.Conclusion：Qingluotongbi granule could inhibit the differentiation and activity of the osteo-like cells,and this action has relation to its inhibiting the overproduction of cytokines in the co-culturing system.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2011年第11期988-992,共5页
Chinese Journal of Immunology
基金
江苏省高校自然基金(08KJB360006)
江苏省六大人才高峰课题(2007)
江苏省中药药效与安全性评价重点实验室开放课题资助(T09004)
