目的 探讨青藤碱通过长链非编码RNA（long non-coding RNA，LncRNA）核富集转录本1（nuclear-enriched autosomal transcript 1，NEAT1）/高迁移率族蛋白B1（high mobility group box-1 protein，HMGB1）-核因子-κB p65（nuclear factor-κB p65，NF-κB p65）途径改善胶原诱导性关节炎（collagen-induced arthritis，CIA）大鼠炎症反应的免疫学机制。方法 24只雌性Wistar大鼠随机分为对照组、模型组、甲氨蝶呤（0.9 mg/kg）组和青藤碱（100 mg/kg）组，每组6只。除对照组外，其余大鼠构建CIA模型，造模成功后，连续给药4周。检测各组大鼠关节炎评分、足趾容积及血清中肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-1β（interleukin-1β，IL-1β）、IL-6、IL-17A、IL-10、IL-13水平；检测大鼠踝关节骨微结构；检测大鼠肝、肾功能指标及肝、肾、脾、关节病理学变化；qRT-PCR和Western blotting检测大鼠关节中LncRNA NEAT1、HMGB1、Toll样受体4（Toll-like receptor 4，TLR4）、NF-κB p65 mRNA和蛋白表达水平；免疫组化法分析大鼠关节中HMGB1、TLR4、NF-κB p65蛋白表达及定位。结果 与对照组比较，模型组大鼠踝关节肿胀明显，足趾容积显著升高（P＜0.05、0.01、0.001），血清中TNF-α、IL-1β、IL-6、IL-17A水平显著升高（P＜0.001），IL-10、IL-13水平显著降低（P＜0.001）；踝关节骨侵蚀严重，骨密度（bone mineral density，BMD）、骨体积分数（bone volume fraction，BV/TV）、骨小梁厚度（trabecular thickness，Tb.Th）显著降低（P＜0.001），骨小梁分离度（trabecular separation，Tb.Sp）显著升高（P＜0.01）；脾脏淋巴小结大小及数量显著增加，关节腔缩小并且有大量炎性细胞浸润，软骨组织破坏；关节中LncRNA NEAT1、HMGB1、TLR4、NF-κB p65 mRNA表达水平显著升高（P＜0.001），HMGB1、TLR4、NF-κB p65蛋白表达水平显著升高（P＜0.05、0.01、0.001）。与模型组比较，各给药组大鼠踝关节肿胀程度明显减轻，关节炎评分、足趾容积显著降低（P＜0.05、0.01、0.001），血清中TNF-α、IL-1β、IL-6、IL-17A水平显著降低（P＜0.001），IL-10、IL-13水平显著升高（P＜0.001）；踝关节表面侵蚀情况有不同程度的缓解，BMD、BV/TV、Tb.Th显著升高（P＜0.01、0.001），Tb.Sp显著降低（P＜0.05、0.01）；肝、肾功能指标及病理学检测正常；淋巴小结数量减少，关节、软骨破坏情况明显减轻；关节中LncRNA NEAT1、HMGB1、TLR4、NF-κB p65 mRNA表达水平显著降低（P＜0.001），HMGB1、TLR4、NF-κB p65蛋白表达水平显著降低（P＜0.05、0.01、0.001）。结论 青藤碱通过调控LncRNA NEAT1/HMGB1-NF-κB p65途径，减轻关节肿胀和损伤，抑制炎症反应，缓解类风湿关节炎进程。

Objective To investigate the immunological mechanism of sinomenine in improving inflammatory response in rats with collagen-induced arthritis (CIA) through long non-coding RNA nuclear-enriched autosomal transcript 1 (LncRNA NEAT1)/high mobility group box-1 protein (HMGB1)-nuclear factor-κB p65 (NF-κB p65) pathway. Methods A total of 24 female Wistar rats were randomly divided into control group, model group, methotrexate (0.9 mg/kg) group and sinomenine (100 mg/kg) group, with six rats in each group. Except the control group, the other rats were all made CIA model. After successful modeling, the drugs were given continuously for four weeks. The arthritis scores, toe volume, levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6, IL-17A, IL-10 and IL-13 in serum were detected; Ankle joint bone micro-structure was detected; Liver and kidney function indicators, as well as the pathological changes of liver, kidney, spleen and joints of each group of rats were detected; The mRNA and protein expression levels of LncRNA NEAT1, HMGB1, Toll-like receptor (TLR4), NF-κB p65 in joints of rats were detected by qRT-PCR and Western blotting; HMGB1, TLR4, NF-κB p65 protein expressions and localization in joints of rats were analyzed by immunohistochemical method. Results Compared with control group, ankle joint swelling and toe volume in model group were significantly increased (P < 0.05, 0.01, 0.001), levels of TNF-α, IL-1β, IL-6 and IL-17A in serum were significantly increased (P < 0.001), while levels of IL-10 and IL-13 were significantly decreased (P < 0.001); The ankle bone erosion was serious, bone mineral density (BMD), bone volume fraction (BV/TV), trabecular thickness (Tb.Th) were significantly decreased (P < 0.001), and trabecular separation (Tb.Sp) was significantly increased (P < 0.01); The size and number of splenic lymph nodes were significantly increased, joint lumen was shrank and there was a large number of inflammatory cells infiltrated, and cartilage tissue was destroyed; The expressions of LncRNA NEAT1, HMGB1, TLR4 and NF-κB p65 mRNA in joints were significantly increased (P < 0.001), and the expressions of HMGB1, TLR4 and NF-κB p65 proteins were significantly increased (P < 0.05, 0.01, 0.001). Compared with model group, the ankle swelling of rats in each administration group was significantly reduced, the arthritis score and toe volume were significantly decreased (P < 0.05, 0.01, 0.001), levels of TNF-α, IL-1β, IL-6 and IL-17A in serum were significantly decreased (P < 0.001), while the levels of IL-10 and IL-13 were significantly increased (P < 0.001); The surface erosion of ankle joint was alleviated to different degrees, BMD, BV/TV, Tb.Th were significantly increased (P < 0.01, 0.001), and Tb.Sp was significantly decreased (P < 0.05, 0.01); Liver and kidney function indexes and pathological tests were normal; The number of lymph nodules was decreased, and the damage of joints and cartilage was obviously alleviated; The expressions of LncRNA NEAT1, HMGB1, TLR4 and NF-κB p65 mRNA in joints were significantly decreased (P < 0.001), and the expressions of HMGB1, TLR4 and NF-κB p65 proteins were significantly decreased (P < 0.05, 0.01, 0.001). Conclusion Sinomenine regulates LncRNA NEAT1/HMGB1-NF-κB p65 pathway, reduces joint swelling and injury, inhibits inflammatory response, and alleviates the progression of rheumatoid arthritis.

R285.5

国家自然科学基金资助项目（32273089，81904034）；山西省基础研究计划面上项目（202303021211172，202403021221262）；中国博士后科学基金（2022M722004）；山西省中医药科技专项科研课题（2023ZYYC058，2024ZYY2A015，2024ZYY2A019）；山西省卫生健康委科研课题（2019011）