Wang Ye-Fan, Lin Zhen-Yang, Zhang Fei-Xu, Zhou Xin-Yue, Wu Xia, Xiao Xiao, Sun Jun-Jiang, Hua Bao-Lai
School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China.
School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China State Key Laboratory of Bioengineering, East China University of Science and Technology, Shanghai 200237, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2022 Oct;30(5):1549-1556. doi: 10.19746/j.cnki.issn.1009-2137.2022.05.038.
To explore the effect of lenalidomide on human fibroblast-like synovial cells (HFLS) and the therapeutic efficacy on hemophilic arthropathy in hemophilia A mice model.
In vitro, to remodel the inflammatory environment of synovial tissue after hemorrhage, ferric citrate and recombinant TNF-α were added into the cell culture medium of HFLS. Cell Counting Kit-8 (CCK-8), Enzyme-linked immunosorbent assay (ELISA), Quantitative Real-time PCR (RT-qPCR) and flow cytometry were employed for detection of the effects of lenalidomide on the proliferation ability, pro-inflammatory cytokines release and apoptosis of HFLS cells. In vivo, hemophilia arthropathy was remodeled in hemophilia A mice by induction of hemarthrosis. A series of doses of lenalidomide (0.1, 0.3 and 1.0 g/kg) was administrated intra-articularly. Tissues of knee joints were collected on the 14th day after administration, and the protective effect of lenalidomide on arthritis in hemophilia A mice were evaluated by RT-qPCR and histological grading.
In vitro, compared with the untreated control group, lenalidomide could significantly inhibit the proliferation of HFLS cells (P<0.05), and the effect was the most significant when the concentration was 0.01 μmol/L (P<0.001). Compared with the control group, lenalidomide could significantly inhibit the expression levels of TNF-α, IL-1β, IL-6 and IFN-γ in HFLS cells (P<0.05). The flow cytometry results showed that lenalidomide could enhance the apoptotis of HFLS cells (P<0.05). The results of RT-qPCR showed that lenalidomide could significantly reduce the mRNA expression levels of TNF-α, IL-1β, IL-6,MCP-1 and VEGF in the joint tissues (P<0.05). Histological results showed that compared with the injured group, lenalidomide could significantly reduce the pathological sequela after hemarthrosis induction, e.g. synovial thickening and neo-angiogenesis in the synovium. The protection displayed a dose-response pattern roughly.
In vitro, lenalidomide can inhibit the proliferation of HFLS cells, promote their apoptosis, and inhibit the expression of pro-inflammatory cytokines. In vivo, lenalidomide can significantly decrease the expression of pro-inflammatory cytokines in the joints of mice, and prevent the development of inflammation and neo-angiogenesis. The results provide a theoretical and experimental basis for the clinical application of lenalidomide in the treatment of hemophilic arthropathy.
探讨来那度胺对人成纤维样滑膜细胞（HFLS）的作用以及对血友病A小鼠模型中血友病性关节病的治疗效果。
体外实验中，为模拟出血后滑膜组织的炎症环境，在HFLS细胞培养基中加入柠檬酸铁和重组TNF-α。采用细胞计数试剂盒-8（CCK-8）、酶联免疫吸附测定（ELISA）、定量实时聚合酶链反应（RT-qPCR）和流式细胞术检测来那度胺对HFLS细胞增殖能力、促炎细胞因子释放及凋亡的影响。体内实验中，通过诱导关节积血在血友病A小鼠中建立血友病性关节病模型。关节腔内给予一系列剂量的来那度胺（0.1、0.3和1.0 g/kg）。给药后第14天收集膝关节组织，通过RT-qPCR和组织学分级评估来那度胺对血友病A小鼠关节炎的保护作用。
体外实验中，与未处理的对照组相比，来那度胺可显著抑制HFLS细胞的增殖（P<0.05），当浓度为0.01 μmol/L时作用最为显著（P<0.001）。与对照组相比，来那度胺可显著抑制HFLS细胞中TNF-α、IL-1β、IL-6和IFN-γ的表达水平（P<0.05）。流式细胞术结果显示来那度胺可增强HFLS细胞的凋亡（P<0.05）。RT-qPCR结果显示来那度胺可显著降低关节组织中TNF-α、IL-1β、IL-6、MCP-1和VEGF的mRNA表达水平（P<0.05）。组织学结果显示，与损伤组相比，来那度胺可显著减轻关节积血诱导后的病理后遗症，如滑膜增厚和滑膜新生血管形成。保护作用大致呈现剂量反应模式。
体外实验中，来那度胺可抑制HFLS细胞的增殖，促进其凋亡，并抑制促炎细胞因子的表达。体内实验中，来那度胺可显著降低小鼠关节中促炎细胞因子的表达，并预防炎症和新生血管形成的发展。这些结果为来那度胺在血友病性关节病治疗中的临床应用提供了理论和实验依据。
