Jiang Yin-Jing, Cheng Yong-Hong, Zhu Hao-Qing, Wu Yan-Ling, Nan Ji-Xing, Lian Li-Hua
Key Laboratory of Traditional Chinese Korean Medicine Research of State Ethnic Affairs Commission, College of Pharmacy, Yanbian University, Yanji, Jilin Province, 133002, China; Key Laboratory of Natural Medicines of the Changbai Mountain, Ministry of Education, College of Pharmacy, Yanbian University, Yanji, Jilin Province, 133002, China.
Key Laboratory of Traditional Chinese Korean Medicine Research of State Ethnic Affairs Commission, College of Pharmacy, Yanbian University, Yanji, Jilin Province, 133002, China; Key Laboratory of Natural Medicines of the Changbai Mountain, Ministry of Education, College of Pharmacy, Yanbian University, Yanji, Jilin Province, 133002, China.
J Ethnopharmacol. 2025 Jan 31;340:119231. doi: 10.1016/j.jep.2024.119231. Epub 2024 Dec 17.
Palmatine (Pal), derived from Daemonorops margaritae (Hance) Becc and Phellodendron amurense Rupr. is a natural isoquinoline alkaloid widely used in clearing heat and drying dampness, purging the pathogenic fire and removing symptoms, detoxifying toxins and healing sores.
Gout is a common metabolic inflammatory disease caused by the deposition of MSU crystals (MSU) in joints and non-articulation structures. Given the multiple toxic side effects of clinical anti-gout medications, there is a need to find a safe and effective alternative. We investigated the therapeutic effects of Pal on MSU crystal-induced acute gouty inflammation, targeting the NLRP3 inflammasome mediated pyroptosis.
In vitro, mouse peritoneal macrophages (MPM) and rat articular chondrocytes were stimulated with LPS plus MSU in the presence or absence of Palmatine. In vivo, arthritis models include the acute gouty arthritis model by injecting MSU crystals in the paws of mice and the air pouch acute gout model by injecting MSU crystals into the mouse subcutaneous tissue of the back. Expression of NLRP3 inflammasome activation and NETosis formation was determined by Western blot, ELISA kit, immunohistochemistry, and immunofluorescence. In addition, the anti-cartilage damage of Palmatine on MSU-induced arthritis mice were also evaluated.
Pal dose-dependently decreased levels of NLRP3 inflammasome activation related proteins NLRP3, ASC, caspase-1, IL-1β, HMGB1 and Cathepsin B. The NETosis protein levels of caspase-11, histone3, PR3 and PAD4 were remarkably reduced by Pal. Pal effectively blocked the activation of NLRP3 inflammasome, attenuated the caspase-11 mediated noncanonical NLRP3 inflammasome activation and intervened the formation of NETs, thereby inhibiting the pyroptosis. In vivo, Pal attenuated MSU-induced inflammation in gouty arthritis and protect the articular cartilage through inhibiting the pyroptosis of proteins NLRP3, ASC, caspase-1, IL-1β, HMGB1 and Cathepsin B, reducing levels of NETosis relevant proteins caspase-11, histone3, PR3 and PAD4 and up-regulating expression of protein MMP-3.
Palmatine ameliorated gouty inflammation by inhibiting pyroptosis via NLRP3 inflammasome.
巴马汀（Pal）来源于麒麟竭（Daemonorops margaritae (Hance) Becc）和黄柏（Phellodendron amurense Rupr.），是一种天然异喹啉生物碱，广泛用于清热燥湿、泻火解毒、解毒消肿。
痛风是一种常见的代谢性炎症性疾病，由单钠尿酸盐（MSU）晶体沉积在关节和非关节结构中引起。鉴于临床抗痛风药物存在多种毒副作用，需要寻找一种安全有效的替代药物。我们研究了巴马汀对MSU晶体诱导的急性痛风性炎症的治疗作用，靶点为NLRP3炎性小体介导的细胞焦亡。
在体外，在有或没有巴马汀的情况下，用脂多糖（LPS）加MSU刺激小鼠腹腔巨噬细胞（MPM）和大鼠关节软骨细胞。在体内，关节炎模型包括通过向小鼠爪部注射MSU晶体建立的急性痛风性关节炎模型和通过向小鼠背部皮下组织注射MSU晶体建立的气袋急性痛风模型。通过蛋白质免疫印迹法、酶联免疫吸附测定试剂盒、免疫组织化学和免疫荧光法测定NLRP3炎性小体激活和中性粒细胞胞外陷阱（NETosis）形成的表达。此外，还评估了巴马汀对MSU诱导的关节炎小鼠的抗软骨损伤作用。
巴马汀剂量依赖性地降低了NLRP3炎性小体激活相关蛋白NLRP3、凋亡相关斑点样蛋白（ASC）、半胱天冬酶-1（caspase-1）、白细胞介素-1β（IL-1β）、高迁移率族蛋白B1（HMGB1）和组织蛋白酶B的水平。巴马汀显著降低了caspase-11、组蛋白3、蛋白酶3（PR3）和瓜氨酸化酶4（PAD4）的NETosis蛋白水平。巴马汀有效阻断了NLRP3炎性小体的激活，减弱了caspase-11介导的非经典NLRP3炎性小体激活，并干预了NETs的形成，从而抑制了细胞焦亡。在体内，巴马汀通过抑制蛋白NLRP3、ASC、caspase-1、IL-1β、HMGB1和组织蛋白酶B的细胞焦亡，降低NETosis相关蛋白caspase-11、组蛋白3、PR3和PAD4的水平，并上调基质金属蛋白酶-3（MMP-3）蛋白的表达，减轻痛风性关节炎中MSU诱导的炎症并保护关节软骨。
巴马汀通过抑制NLRP3炎性小体介导的细胞焦亡改善痛风性炎症。
