骨髓间充质干细胞对类风湿关节炎小鼠干预效果研究

Effect of bone marrow mesenchymal stem cells on rheumatoid arthritis in mice

目的利用自发类风湿关节炎小鼠模型(Tg-huTNFα小鼠),探讨骨髓间充质干细胞(BMSCs)对类风湿关节炎及其继发骨量丢失的影响。方法3周龄Tg-huTNFα小鼠分为3组:(1)模型组(Model),每周腹腔注射1次0.1 mL/10 g的生理盐水;(2)huTNFα抗体干预组(Anti-huTNFα),每周腹腔注射1次10 mg/kg的huTNFα抗体;(3)BMSCs干预组(BMSCs),每周双侧膝关节腔各注射1次5×10^(6) Cells的BMSCs。干预期间每周称量1次动物体重,同时对踝关节进行临床评分,各组动物干预10周后处死取材。利用流式细胞仪分析外周血T细胞、B细胞、CD4^(+)/CD8^(+)T细胞比例,液相芯片法检测血清中huTNFα、IL-4及IL-1β含量,HE染色后评价左侧膝关节和踝关节病理改变,骨形态计量学方法分析右侧股骨骨结构变化。结果与模型组比较,BMSCs和Anti-huTNFα干预均可缓解Tg-huTNFα小鼠体重的下降,抑制小鼠关节肿大变形,下调外周血huTNFα含量,降低Tg-huTNFα小鼠的关节病理评分,减少关节滑膜和软骨损伤面积。此外,骨形态计量学分析发现,BMSCs干预可提高骨小梁数量和骨小梁面积百分数等,改善Tg-huTNFα小鼠股骨骨结构。结论关节腔注射BMSCs可以显著缓解Tg-huTNFα小鼠类风湿关节炎症状,改善股骨骨结构,缓解RA继发的骨量丢失。

Objective To investigate the effect of bone marrow mesenchymal stem cells(BMSCs)on rheumatoid arthritis and secondary bone loss in a spontaneous rheumatoid arthritis mouse model(Tg⁃huTNFαmice).Methods 3⁃week⁃old Tg⁃huTNFαmice were divided into three groups:(1)Model group:0.1 mL/10 g normal saline was intraperitoneally injected once a week;(2)huTNFαantibody intervention group(Anti⁃huTNFα):10 mg/kg Anti⁃hu TNFαwas intraperitoneally injected once a week;(3)BMSC intervention group(BMSCs):5×10^(6) BMSCs were injected into both knee joint cavities once a week.During the intervention period,the animals were weighed once a week,and ankle joint health was scored clinically.The animals were killed after 10 weeks of intervention.The percentages of T cells,B cells and CD4^(+)/CD8^(+)T cells in the peripheral blood were analyzed by flow cytometry.The amounts of huTNFα,IL⁃4 and IL⁃1βin peripheral blood serum were detected by liquid chip technique.Pathological changes to the left knee joint and ankle joint were evaluated by HE staining,and the right femur bone structure was analyzed by bone histomorphometry.Results BMSCs and Anti⁃huTNFαprovided relief from weight loss in Tg⁃huTNFαmice;inhibited joint swelling and deformation;and reduced the peripheral blood huTNFαcontent,joint pathology score,and extent of joint synovium and cartilage damage.Additionally,BMSCs improved the trabecular bone number and percentage trabecular bone area to different degrees and improved the femur bone structure in Tg⁃huTNFαmice.Conclusions Injection of BMSCs into the joint cavity significantly alleviated symptoms of rheumatoid arthritis in Tg⁃huTNFαmice,improved femur bone structure,and alleviated bone loss secondary to RA.