Sichuan Industrial Institute of Antibiotics, School of Pharmacy, Chengdu university, Chengdu 610106, China.
School of Food and Biological engineering, Chengdu university, Chengdu 610106, China.
Phytomedicine. 2022 Sep;104:154339. doi: 10.1016/j.phymed.2022.154339. Epub 2022 Jul 13.
As a prolonged autoimmune disorder, rheumatoid arthritis (RA) is characterised by synovial hyperplasia and the erosion of bone and cartilage. Magnoflorine (MAG) is the main component purified from Clematis manshurica Rupr. Recent studies have shown that MAG has anti-inflammatory, antioxidant, and immunosuppressive effects, which are relevant to anti-RA activities.
The current investigation was conducted to explore the anti-RA effects of MAG and to discover the possible molecular mechanisms.
In vitro experiments, CCK-8, wound healing, and transwell assays were utilized to evaluate the anti-proliferative, anti-migratory, and anti-invasive activities of MAG, respectively. The rate of cell distribution and cell apoptosis were evaluated by flow cytometry. ROS generation was detected by DCFH-DA staining. Western blotting, quantitative real-time polymerase chain reaction assay, and immunofluorescent staining were employed to test the anti-RA effect of MAG as well as to explore the potential mechanisms by evaluating related gene and protein expression. For in vivo experiments, an adjuvant-induced arthritis (AIA) rat model was established. The related parameters were measured in rats. Then, rats were sacrificed, and ankle joints were collected for histopathological analysis and observation.
MAG significantly decreased the proliferation, migration, invasion, and reactive oxygen species levels in IL-1β-treated MH7A cells. Furthermore, MAG promoted cell apoptosis by increasing Bax levels and decreasing Bcl-2 levels. MAG also induced cell cycle arrest. Inflammatory cytokines (iNOS, COX-2, IL-6, and IL-8) and MMPs (MMP-1, 2, 3, 9, and 13) were reduced by MAG treatment. Molecular analysis revealed that MAG exerted anti-RA effects by partly inhibiting the PI3K/Akt/NF-κB signalling axis and activating the Keap1-Nrf2/HO-1 signalling pathway. In vivo studies have revealed that MAG treatment substantially improved severe symptoms in AIA rats, and these curative effects were linked to the attenuation of inflammatory responses.
These results first suggested that MAG exhibits anti-arthritic effects in IL-1β-treated MH7A cells and AIA rat models. Thus, MAG may be used as a new drug to treat RA clinically.
类风湿关节炎（RA）作为一种长期的自身免疫性疾病，其特征为滑膜增生以及骨和软骨的侵蚀。蝙蝠葛碱（MAG）是从威灵仙中提取的主要成分。最近的研究表明，MAG 具有抗炎、抗氧化和免疫抑制作用，与抗 RA 活性有关。
本研究旨在探讨 MAG 的抗 RA 作用及其可能的分子机制。
在体外实验中，CCK-8、划痕愈合和 Transwell 实验分别用于评估 MAG 的抗增殖、抗迁移和抗侵袭活性。通过流式细胞术评估细胞分布和细胞凋亡率。通过 DCFH-DA 染色检测 ROS 的产生。Western blot、实时定量聚合酶链反应（qPCR）和免疫荧光染色用于测试 MAG 的抗 RA 作用，并通过评估相关基因和蛋白表达来探讨其潜在机制。对于体内实验，建立了佐剂诱导关节炎（AIA）大鼠模型。测量大鼠的相关参数。然后处死大鼠，收集踝关节进行组织病理学分析和观察。
MAG 显著降低了 IL-1β 处理的 MH7A 细胞的增殖、迁移、侵袭和活性氧水平。此外，MAG 通过增加 Bax 水平和降低 Bcl-2 水平促进细胞凋亡。MAG 还诱导细胞周期停滞。炎性细胞因子（iNOS、COX-2、IL-6 和 IL-8）和 MMPs（MMP-1、2、3、9 和 13）的表达被 MAG 处理所抑制。分子分析表明，MAG 通过部分抑制 PI3K/Akt/NF-κB 信号通路和激活 Keap1-Nrf2/HO-1 信号通路发挥抗 RA 作用。体内研究表明，MAG 治疗可显著改善 AIA 大鼠的严重症状，这些疗效与炎症反应的减弱有关。
这些结果首次表明，MAG 在 IL-1β 处理的 MH7A 细胞和 AIA 大鼠模型中具有抗关节炎作用。因此，MAG 可能被用作临床上治疗 RA 的新药。
