Department of Anatomy, School of Basic Medical Sciences, Hubei University of Chinese Medicine, Wuhan, China.
College of Acupuncture and Moxibustion, Hubei University of Chinese Medicine, Wuhan, China.
Immunopharmacol Immunotoxicol. 2020 Apr;42(2):147-155. doi: 10.1080/08923973.2020.1732407. Epub 2020 Mar 2.
Sinomenine (SIN), an alkaloid isolated from plant, possesses many pharmacological properties, such as anti-inflammation, anti-hyperalgesia, anti-allergy, anti-apoptosis, and anti-angiogenesis. In this study, we aimed to investigate the detailed molecular mechanisms associated with the anti-inflammatory activity of SIN in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. RAW264.7 cells were treated with LPS and/or indicated concentrations of SIN. Inflammatory cytokine production, such as TNF-α, IL-1β, and IL-6, was detected by ELISA. Expression of microRNA-155 (miR-155), SOCS1 and NF-κB was assessed by qRT-PCR and Western blot, separately. Simultaneously, miR-155 inhibitor and SOCS1 SiRNA were transfected to observe the regulative effects of SIN on the expression of miR-155, SOCS1, and NF-κB. Our result showed that SIN treatment significantly reduced LPS-induced inflammatory cytokine release, suppressed the expression of miR-155, enhanced SOCS1 expression at mRNA and protein levels, and prevented NF-ĸB transcription. Furthermore, transfection of miR-155 inhibitor and SOCS1 SiRNA emphasized that the regulation of miR-155, SOCS1, and NF-ĸB was associated with the anti-inflammatory activation of SIN in LPS-treated macrophages. This study indicated that SIN alleviated LPS-induced inflammatory responses in RAW264.7 macrophages by downregulating miR-155 and upregulating SOCS1, at least partly, leading to the suppression of NF-ĸB transcription. These findings suggest that SIN might be developed as an alternative and promising drug for the treatment of inflammatory diseases.
青藤碱（SIN）是从植物中分离得到的一种生物碱，具有多种药理作用，如抗炎、抗痛觉过敏、抗过敏、抗凋亡和抗血管生成。在这项研究中，我们旨在探讨 SIN 抑制脂多糖（LPS）刺激的 RAW264.7 巨噬细胞炎症反应的详细分子机制。用 LPS 和/或不同浓度 SIN 处理 RAW264.7 细胞。通过 ELISA 检测 TNF-α、IL-1β 和 IL-6 等炎症细胞因子的产生。分别通过 qRT-PCR 和 Western blot 检测 microRNA-155（miR-155）、SOCS1 和 NF-κB 的表达。同时，转染 miR-155 抑制剂和 SOCS1 SiRNA，观察 SIN 对 miR-155、SOCS1 和 NF-κB 表达的调节作用。结果表明，SIN 处理可显著降低 LPS 诱导的炎症细胞因子释放，抑制 miR-155 的表达，增强 SOCS1 的表达，同时抑制 NF-κB 的转录。此外，转染 miR-155 抑制剂和 SOCS1 SiRNA 强调了 miR-155、SOCS1 和 NF-κB 的调节与 SIN 在 LPS 处理的巨噬细胞中的抗炎作用有关。本研究表明，SIN 通过下调 miR-155 和上调 SOCS1 缓解 LPS 诱导的 RAW264.7 巨噬细胞炎症反应，从而抑制 NF-κB 转录。这些发现表明，SIN 可能被开发为治疗炎症性疾病的替代和有前途的药物。
