Yang R, Shu Y, Wen H, Cai X, Wang Z, Zhang C, Xiang Y, Wu H
Department of Medicine, Enshi University, Enshi 445000, China.
Hubei Provincial Key Laboratory of Occurrence and Intervention of Rheumatic Diseases, Enshi 445000, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2024 Sep 20;44(9):1645-1652. doi: 10.12122/j.issn.1673-4254.2024.09.03.
To investigate the therapeutic mechanism of Skan total flavonoids (PHSTF) for rheumatoid arthritis (RA).
Twenty-five male SD rats were randomly divided into normal control group, RA model group, PHSTF treatment (45 and 90 mg/kg) groups, and Tripterygium glycosides (TPG) tablet (10 mg/kg) group (=5). Except for those in the normal control group, all the rats were subjected to collagen-induced arthritis (CIA) modeling using a secondary immunization method, after which PHSTF and TPG were administered via gavage once daily for 4 weeks. After the treatments, serum levels of TNF- and IL-1β were measured using ELISA, and ankle joint pathologies were assessed with HE staining; the expression of citrullinated histone H3 (Cit-H3), a neutrophil extracellular trap (NET) marker, in the ankle joints was evaluated with immunohistochemistry. In primary cultures of rat peripheral blood neutrophils stimulated with phorbol ester (PMA), the effects of PHSTF (100 and 200 μg/mL) on the expressions of Cit-H3, peptidylarginine deiminase 4 (PADI4), neutrophil elastase (NE), and myeloperoxidase (MPO) were examined with Western blotting; immunofluorescence assay was used to observe Cit-H3 expression and NET formation in the cells.
In the CIA rat models, PHSTF significantly alleviated ankle swelling, decreased serum levels of TNF- and IL-1β, improved histopathological changes in the ankle joints, and reduced Cit-H3 expression in both the serum and ankle joint cartilage. In the isolated rat neutrophils, PHSTF showed no significant effect on cell viability but strongly inhibited PMA-induced NET release.
PHSTF can alleviate RA by inhibiting the formation of NETs.
探讨光叶菝葜总黄酮（PHSTF）治疗类风湿关节炎（RA）的作用机制。
将25只雄性SD大鼠随机分为正常对照组、RA模型组、PHSTF治疗组（45和90 mg/kg）、雷公藤多苷片（TPG）组（10 mg/kg），每组5只。除正常对照组外，其余大鼠采用二次免疫法建立胶原诱导性关节炎（CIA）模型，造模后PHSTF和TPG灌胃给药，每日1次，共4周。治疗后，采用酶联免疫吸附测定（ELISA）法检测血清肿瘤坏死因子-α（TNF-α）和白细胞介素-1β（IL-1β）水平，苏木精-伊红（HE）染色评估踝关节病理变化；采用免疫组织化学法评估踝关节中瓜氨酸化组蛋白H3（Cit-H3）（一种中性粒细胞胞外诱捕网（NET）标志物）的表达。在佛波酯（PMA）刺激的大鼠外周血中性粒细胞原代培养物中，采用蛋白质免疫印迹法检测PHSTF（100和200 μg/mL）对Cit-H3、肽基精氨酸脱亚氨酶4（PADI4）、中性粒细胞弹性蛋白酶（NE）和髓过氧化物酶（MPO）表达的影响；采用免疫荧光法观察细胞中Cit-H3的表达和NET的形成。
在CIA大鼠模型中，PHSTF显著减轻踝关节肿胀，降低血清TNF-α和IL-1β水平，改善踝关节组织病理学变化，并降低血清和踝关节软骨中Cit-H3的表达。在分离的大鼠中性粒细胞中，PHSTF对细胞活力无显著影响，但强烈抑制PMA诱导的NET释放。
PHSTF可通过抑制NET形成来缓解RA。
