海南医科大学学报

目的：观察雷藤舒对类风湿关节炎小鼠铁死亡的影响及治疗作用，并基于肿瘤蛋白53(p53)/溶质载体家族7成员11(SLC7A11)/谷胱甘肽过氧化物酶4(GPX4)通路探究其可能机制。方法：将40只雄性DBA小鼠，分为Control组、Model组、雷藤舒（LLDT-8）组（0.5 mg/kg）和铁死亡抑制剂（Fer-1）组（1 mg/kg），每组10只。除Control组外，采用Ⅱ型胶原诱导法制备类风湿关节炎（CIA）模型。造模成功后，各给药组进行相应药物干预，Control组和Model组灌胃等体积生理盐水，干预周期为4周。主要观察指标包括：小鼠足趾厚度及关节炎临床评分；微计算机断层扫描评估骨破坏；苏木素-伊红染色和番红O-固绿染色法评估关节组织病理学变化；酶联免疫吸附测定法检测血清中MMP-3、TNF-α、IL-1β和IL-6；比色法检测关节组织中MDA、GSH和NADPH含量；免疫荧光检测关节组织中的ROS；免疫组化法及实时荧光定量PCR检测SLC7A11、GPX4、p53蛋白及mRNA表达。结果：与Control组比较，Model组小鼠关节红肿畸形，关节炎临床评分明显升高，关节组织骨破坏、滑膜增生及炎性细胞浸润，血清MMP-3、TNF-α、IL-1β、IL-6水平明显上升；关节组织GSH、NADPH水平明显升高，ROS、MDA含量明显降低；SLC7A11、GPX4蛋白及mRNA表达均明显上调，p53蛋白及mRNA表达下调，组间差异均有统计学意义（P<0.05）。与Model组比较，LLDT-8组可显著改善小鼠关节肿胀变形，降低关节炎临床积分，减轻小鼠关节骨破坏以及减少炎症细胞浸润和滑膜增生，血清MMP-3、TNF-α、IL-1β、IL-6表达水平明显降低，关节组织GSH、NADPH水平明显降低，ROS、MDA含量明显升高，SLC7A11、GPX4蛋白及mRNA表达均明显下调，p53蛋白及mRNA表达上调，组间差异均有统计学意义（P<0.05）。结论：雷藤舒可有效减轻CIA小鼠滑膜增生及炎性浸润，缓解关节组织病理损伤，其机制或与调控p53/SLC7A11/GPX4信号通路，增加铁死亡敏感性有关。

Objective: To investigate the effects of(5R)-5-Hydroxytriptolide(LLDT-8) on ferroptosis in a murine model of rheumatoid arthritis(RA) and to elucidate its underlying mechanisms via the tumor protein 53(p53)/solute carrier family 7 member 11(SLC7A11)/glutathione peroxidase 4(GPX4) signaling pathway. Methods: A total of 40 male DBA mice were allocated into the Control, the Model,the LLDT-8(0.5 mg/kg), and the Ferroptosis inhibitor(Fer-1)(1 mg/kg)groups, with 10 mice per group. All groups except the Control received collagen type Ⅱ to induce collagen-induced arthritis(CIA). Following the successful modeling, the treatment groups were administered their respective drugs, while the Control and Model groups were gavaged with an equal volume of saline for 4 weeks. The key observation metrics included: the thickness of the mice's toes and clinical scores for arthritis; assessment of bone damage using micro-computed tomography; examination of pathological changes in joint tissues via hematoxylin-eosin staining and Sirius Red-O green staining; measurement of serum levels of MMP-3, TNF-α, IL-1β, and IL-6 using enzyme-linked immunosorbent assay; evaluation of MDA, GSH, and NADPH contents in joint tissues through colorimetric methods; detection of reactive oxygen species(ROS) in joint tissues via immunofluorescence; and analysis of SLC7A11, GPX4, and p53 protein and mRNA expression using immunohistochemistry and real-time quantitative polymerase chain reaction. Results: The Model group exhibited significant joint swelling, deformities, and increased clinical scores, along with pronounced bone destruction, synovial hyperplasia, and inflammatory cell infiltration compared to the Control group. Serum levels of MMP-3, TNF-α, IL-1β, and IL-6 were significantly elevated, while GSH and NADPH levels in joint tissues were markedly increased, and ROS and MDA content were significantly reduced. Notably, SLC7A11 and GPX4 protein and mRNA expressions were significantly upregulated, whereas p53 protein and mRNA expressions were notably downregulated(P<0.05).Compared to the Model group treatment with LLDT-8 led to significant improvements in joint swelling and deformities, a reduction in clinical scores, mitigation of bone destruction, and a decrease in inflammatory cell infiltration and synovial hyperplasia. Additionally, serum levels of MMP-3, TNF-α, IL-1β, and IL-6 were significantly lowered, while GSH and NADPH levels in joint tissues decreased, and ROS and MDA content significantly increased. The protein and mRNA expressions of SLC7A11 and GPX4 were significantly downregulated, whereas p53 protein and mRNA expressions were upregulated(P<0.05). Conclusion: LLDT-8 effectively reduces synovial hyperplasia and inflammatory infiltration in CIA mice, alleviating pathological damage to joint tissues. Its mechanism may involve the modulation of the p53/SLC7A11/GPX4 signaling pathway, thereby enhancing sensitivity to ferroptosis.