文献类型：期刊文献

中文题名：来那度胺对血友病性关节炎的治疗作用研究

英文题名：Study on the Therapeutic Effect of Lenalidomide on Hemophilic Arthropathy

作者：王冶凡[1];林振洋[1];张飞旭[1];周新月[1];吴侠[1];肖啸[1,2];孙君江[1,2];华宝来[3,4]

机构：[1]华东理工大学药学院,上海200237;[2]华东理工大学生物反应器工程国家重点实验室,上海200237;[3]徐州医科大学扬州临床学院,江苏扬州225001;[4]扬州大学临床医学院血液科,江苏扬州225001

年份：2022

卷号：30

期号：5

起止页码：1549

中文期刊名：中国实验血液学杂志

外文期刊名：Journal of Experimental Hematology

收录：CSTPCD;;Scopus;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；PubMed;

语种：中文

中文关键词：来那度胺;血友病性关节炎;人滑膜成纤维细胞;炎性因子;小鼠

外文关键词：lenalidomide;hemophilia arthropathy;human fibroblast-like synovial cells;pro-inflammatory cytokines;mouse

摘要：目的:探讨来那度胺对人滑膜成纤维细胞(HFLS)增殖的影响以及对A型血友病小鼠血友病性关节炎(HA)的治疗效果。方法:以HFLS为模型细胞,在细胞培养基中添加柠檬酸铁和TNF-α重组蛋白,体外模拟炎性环境,加入不同浓度的来那度胺,通过CCK-8、ELISA、RT-q PCR、流式细胞术等方法检测HFLS的增殖能力、促炎细胞因子释放和细胞凋亡等功能的变化;构建小鼠HA模型,关节腔内注射不同浓度(0.1、0.3和1.0 g/kg)的来那度胺,在给药后d 14收集样本,通过RT-q PCR检测和组织学方法观察小鼠关节中滑膜增厚、新生血管形成等情况。结果:在体外,与未加药物的对照组相比,来那度胺0.001、0.01、0.1、1、10μmol/L均能够显著抑制HFLS的增殖活力(P<0.05),以浓度为0.01μmol/L时作用最显著(P<0.001)。与对照组相比,来那度胺能够显著抑制HFLS细胞TNF-α、IL-1β、IL-6和IFN-γ等炎性因子的表达水平(P<0.05)。流式细胞术检测结果表明,来那度胺能够促进HFLS细胞的凋亡水平(P<0.05)。RT-q PCR结果表明来那度胺能够显著降低小鼠关节组织中TNF-α、IL-1β、IL-6、MCP-1和VEGF的mRNA表达水平(P<0.05)。组织学病理检测结果表明,来那度胺能够明显降低HA小鼠的关节滑膜厚度,减少新生血管生成,效果呈现剂量依赖性(r=-0.9129),以剂量为1 g/kg时保护作用最明显。结论:在体外,来那度胺能够有效抑制HFLS细胞增殖,促进其凋亡,并抑制其炎性因子的表达;在体内,能够缓解小鼠关节内炎症发展,显著降低小鼠关节中炎性因子的表达水平,为来那度胺将来应用于HA的临床治疗提供了理论和实验依据。
Objective: 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. Methods: 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. Results: 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 RTqPCR 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. Conclusion: In vitro,lenalidomide can inhibit the proliferation of HFLS cells, promote their apoptosis, and inhibit the expression of proinflammatory 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.