School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, 510006, People's Republic of China.
The Second Clinical College of Guangzhou University of Chinese Medicine, Guangzhou, 510120, People's Republic of China.
Drug Des Devel Ther. 2022 Jul 2;16:2119-2132. doi: 10.2147/DDDT.S356307. eCollection 2022.
Gouty arthritis could be triggered by the deposition of monosodium uric acid (MSU) crystals. Palmatine (PAL), a protoberberine alkaloid, has been proven to possess compelling health-beneficial activities. In this study, we aimed to explore the effect of PAL on LPS plus MSU crystal-stimulated gouty arthritis in vitro and in vivo.
PMA-differentiated THP-1 macrophages were primed with LPS and then stimulated with MSU crystal in the presence or absence of PAL. The expression of pro-inflammatory cytokines and oxidative stress-related biomarkers and signal pathway key targets were determined by ELISA kit, Western blot, immunohistochemistry and qRT-PCR, respectively. In addition, the anti-inflammatory and antioxidant activities of PAL on MSU-induced arthritis mice were also evaluated.
The results indicated that PAL (20, 40 and 80 μM) dose-dependently decreased the mRNA expression and levels of pro-inflammatory cytokines (interleukin-1beta (IL-1β), IL-6, IL-18 and tumor necrosis factor alpha (TNF-α)). The levels of superoxide dismutase (SOD) and glutathione (GSH) were remarkably enhanced, while the level of malondialdehyde (MDA) was reduced. Western blot analysis revealed that PAL appreciably inhibited NF-κB/NLRP3 signaling pathways through inhibiting the phosphorylation of p-65 and IκBα, blocking the expression of NLRP3, ASC, IL-1β and Caspase-1, as well as enhancing the antioxidant protein expression of Nrf2 and HO-1. In vivo, PAL attenuated MSU-induced inflammation in gouty arthritis, as evidenced by mitigating the joint swelling, and decreasing the productions of IL-1β, IL-6, IL-18, TNF-α and MDA, while enhancing the levels of SOD and GSH. Moreover, PAL further attenuated the infiltration of neutrophils into joint synovitis.
PAL protected against MSU-induced inflammation and oxidative stress via regulating the NF-κB/NLRP3 and Nrf2 pathways. PAL may represent a potential candidate for the treatment of gouty arthritis.
尿酸单钠（MSU）晶体的沉积可引发痛风性关节炎。黄连碱（PAL）是一种原小檗碱生物碱，已被证明具有令人信服的健康有益活性。在这项研究中，我们旨在探索 PAL 对 LPS 加 MSU 晶体刺激体外和体内痛风性关节炎的影响。
PMA 分化的 THP-1 巨噬细胞先用 LPS 预处理，然后在存在或不存在 PAL 的情况下用 MSU 晶体刺激。通过 ELISA 试剂盒、Western blot、免疫组化和 qRT-PCR 分别测定促炎细胞因子和氧化应激相关生物标志物以及信号通路关键靶标的表达。此外，还评估了 PAL 对 MSU 诱导的关节炎小鼠的抗炎和抗氧化活性。
结果表明，PAL（20、40 和 80 μM）剂量依赖性地降低了促炎细胞因子（白细胞介素-1β（IL-1β）、IL-6、IL-18 和肿瘤坏死因子-α（TNF-α））的 mRNA 表达和水平。超氧化物歧化酶（SOD）和谷胱甘肽（GSH）的水平显著增强，而丙二醛（MDA）的水平降低。Western blot 分析表明，PAL 通过抑制 p-65 和 IκBα 的磷酸化，阻断 NLRP3、ASC、IL-1β 和 Caspase-1 的表达，以及增强 Nrf2 和 HO-1 的抗氧化蛋白表达，显著抑制 NF-κB/NLRP3 信号通路。体内，PAL 减轻了 MSU 诱导的痛风性关节炎中的炎症，表现为减轻关节肿胀，降低 IL-1β、IL-6、IL-18、TNF-α 和 MDA 的产生，同时提高 SOD 和 GSH 的水平。此外，PAL 进一步减轻了中性粒细胞浸润到关节滑膜炎。
PAL 通过调节 NF-κB/NLRP3 和 Nrf2 途径来保护 MSU 诱导的炎症和氧化应激。PAL 可能是治疗痛风性关节炎的潜在候选药物。
