LLDT-8对类风湿关节炎单核细胞向破骨样细胞转化的作用机制研究  

Study of the mechanism of Leitengshu on the transformation of rheumatoid arthritis monocytes into osteoclast like cells

作　　者：严雅 何东仪 聂红[3] 冯伟[4] 沈逸 姜婷 连艳玲 丁琴 YAN Ya;HE Dongyi;NIE Hong;FENG Wei;SHEN Yi;JIANG Ting;LIAN Yanling;DING Qin(General Practice Department of Wujing Community Health Service Center of Minhang District,Shanghai 200241,China;Department of Arthritis of Guanghua Hospital of Integrated Traditional Chinese and Western Medicine Hospital,Shanghai 200052,China;Shanghai Institute of Immunology of Medical School of Shanghai Jiao Tong University,Shanghai 200025,China;Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Tianlin Community Health Service Center of Xiuhui District,Shanghai 200032,China)

机构地区：[1]上海闵行区吴泾社区卫生服务中心全科,上海200241 [2]上海市光华中西医结合医院关节内科,上海200052 [3]上海交通大学医学院上海市免疫研究所,上海200025 [4]上海中医药大学,上海201203 [5]上海市徐汇区田林街道社区卫生服务中心,上海200032

出　　处：《上海医药》2022年第8期19-24,41,共7页Shanghai Medical & Pharmaceutical Journal

摘　　要：目的:探讨LLDT-8对类风湿关节炎骨破坏的作用机制。方法:收集取类风湿关节炎患者的外周血和关节滑液,分离外周血单个核细胞(PBMC)和关节滑液单个核细胞(SFMC)。PBMC用RANKL和M-CSF分别诱导后加入不同浓度LLDT-8干预,于24、48、72 h和14 d进行抗酒石酸酸性磷酸酶染色,显微镜下对Trap(+)细胞计数,同时收集上清检测RANKL、RANK、OPG,比较组间差异。PBMC、SFMC用不同浓度LLDT-8干预后,用ELISA方法检测不同标本中RANKL、OPG、VEGF的分泌,另外用实时PCR检测不同标本PBMC中RANKL、OPG、VEGF、MMP-9的表达,以了解LLDT-8对骨破坏相关因子的影响。结果:LLDT-8减少PBMC向破骨样细胞的转化,促进破骨样细胞的凋亡;LLDT-8抑制VEGF的表达,上调OPG mRNA表达,同时下调VEGF、RANKL和MMP-9的表达。结论:LLDT-8促进破骨样细胞的凋亡,抑制OC的分化,降低RANKL/OPG比值,下调VEGF的表达,延缓骨破坏。Objective:To explore the mechanism of Leitengshu(LLDT-8)on bone destruction in rheumatoid arthritis.Methods:Peripheral blood and synovial fluid were collected from patients with rheumatoid arthritis,Peripheral blood mononuclear cells(PBMC)and synovial fluid mononuclear cells(SFMC)were isolated.PBMCs were induced with RANKL and M-CSF,respectively,and then intervened with different concentrations of LLDT-8,tartrate-resistant acid phosphatase staining was performed at 24,48,72 h and 14 d,and Trap(+)cells were counted under a microscope,at the same time,the supernatant was collected to detect RANKL,RANK,and OPG,and the differences between groups were compared.After PBMC and SFMC were intervened with different concentrations of LLDT-8,the secretion of RANKL,OPG and VEGF in different samples was detected by Elisa method,in addition,real-time PCR was used to detect the expressions of RANKL,OPG,VEGF and MMP-9 in PBMC of different specimens to understand the effect of LLDT-8 on bone destruction-related factors.Results:LLDT-8 reduced the transformation of PBMCs into osteoclast-like cells and promoted the apoptosis of osteoclast-like cells;LLDT-8 inhibited the expression of VEGF,up-regulated the expression of OPG mRNA,and down-regulated the expression of VEGF,RANKL and MMP-9.Conclusion:LLDT-8 promotes the apoptosis of osteoclast-like cells,inhibits the differentiation of OC,decreases the RANKL/OPG ratio,down-regulates the expression of VEGF,and delay bone destruction.

关 键 词：类风湿关节炎 LLDT-8 骨破坏 细胞因子 

分 类 号：R2[医药卫生—中医学]