Magnoflorine attenuates inflammatory responses in RA by regulating the PI3K/Akt/NF-κB and Keapl-Nrf2/HO-1 signalling pathways in vivo and in vitro
Magnoflorine attenuates inflammatory responses in RA by regulating the PI3K/Akt/NF-κB and Keapl-Nrf2/HO-1 signalling pathways in vivo and in vitro
复制标题
Magnoflorine 通过调节体内和体外 PI3K/Akt/NF-κB 和 Keap1-Nrf2/HO-1 信号通路来减轻 RA 炎症反应
DOI：
10.1016/j.phymed.2022.154339
复制
发表时间：
2022-07-20
期刊：
影响因子：
7.9
通讯作者：
Fu, Qiang
中科院分区：
文献类型：
--
作者：
Shen, Yue;Fan, Xinting;Fu, Qiang
背景：类风湿关节炎(RA)是一种长期的自身免疫性疾病，以滑膜增生和骨、软骨侵蚀为特征。木兰花碱(Magnoflorine，MAG)是从东北铁线莲中分离纯化的主要成分。近年来的研究表明，MAG具有抗炎、抗氧化和免疫抑制作用，这些作用与其抗RA活性有关。目的：探讨MAG的抗RA作用及其可能的分子机制。方法：采用体外实验、CCK-8法、创面愈合实验和Transwell实验分别评价MAG的抗增殖、抗迁移和抗侵袭活性。流式细胞仪检测细胞分布率和细胞凋亡率。DCFH-DA染色检测ROS的产生。用Western blotting、实时定量聚合酶链式反应和免疫荧光染色检测MAG的抗RA作用，并通过检测相关基因和蛋白的表达来探讨其可能的作用机制。对于体内实验，建立了佐剂性关节炎(AIA)大鼠模型。以大鼠为实验动物，测量相关参数。结果：MAG能显著降低IL-1β诱导的MH7A细胞的增殖、迁移、侵袭和活性氧水平。此外，MAG还通过提高Bax水平和降低Bcl2水平来促进细胞凋亡。MAG还可诱导细胞周期停滞。MAG可降低炎性细胞因子(iNOS、COX-2、IL-6、IL-8)和MMPs(MMP1、2、3、9、13)。分子分析表明MAG通过部分抑制PI3K/Akt/NF-kappa B信号轴和激活Keap1-Nrf2/HO-1信号通路发挥抗RA作用。体内研究表明，MAG可显著改善AIA大鼠的严重症状，这些疗效与减轻炎症反应有关。结论：MAG对IL-1β诱导的MH7A细胞和AIA大鼠模型具有抗关节炎作用。因此，MAG有可能成为临床治疗类风湿关节炎的新药。
Background: 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.Objective: The current investigation was conducted to explore the anti-RA effects of MAG and to discover the possible molecular mechanisms.Methods: In vitro experiments, CCK-8, wound healing, and transwell assays were utilized to evaluate the antiproliferative, 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.Results: MAG significantly decreased the proliferation, migration, invasion, and reactive oxygen species levels in IL-1 beta-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-kappa B signalling axis and activating the Keapl-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.Conclusion: These results first suggested that MAG exhibits anti-arthritic effects in IL-1 beta-treated MH7A cells and AIA rat models. Thus, MAG may be used as a new drug to treat RA clinically.