Guan Yin, Zhao Xiaoqian, Liu Weiwei, Wang Yue
Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, 210029, Jiangsu, China.
Department of Ethics Committee, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, 210029, Jiangsu, China.
J Orthop Surg Res. 2020 Oct 21;15(1):484. doi: 10.1186/s13018-020-02004-x.
Galuteolin (Galu) is a substance extracted and purified from honeysuckle. The purpose of this study was to explore the effects of Galu on the TNF-α-induced RA-FLS cells (synoviocytes) and reveal its potential molecular mechanism from the perspectives of anti-apoptosis and anti-inflammation.
After TNF-α stimulation, cell proliferation of RA-FLS was assessed by CCK-8 assay. TUNEL staining was used to detect the apoptosis. Western blot was used to detect the expressions of Iκκβ, p-p65, p65, p-IκB, IκB, Cleaved-caspase3, Caspase-3, Bcl-2, and Bax. HO-1 were determined by RT-PCR. The contents of pro-inflammatory cytokines IL-1β, IL-6, IL-8, and MMP-1 were determined by ELISA.
Galu significantly suppressed cell proliferation in a dose-dependent manner. Additionally, Galu obviously promotes cell apoptosis rate of RA-FLS cells and elevated the expression levels of HO-1, caspase-3, and Bax, while reducing the expression level of Bcl-2. Furthermore, Galu apparently inhibited the levels of Iκκβ, p-p65, and p-IκB. Moreover, Galu also significantly reduced the levels of pro-inflammatory factors IL-1β, IL-6, IL-8, and MMP-1 in RA-FLS cells.
Galuteolin exerts protective effects against TNF-α-induced RA-FLS cells by inhibiting apoptosis and inflammation, which can guide the clinical use of rheumatoid arthritis.
木犀草素（Galu）是从金银花中提取纯化的一种物质。本研究旨在探讨Galu对肿瘤坏死因子-α（TNF-α）诱导的类风湿关节炎成纤维样滑膜细胞（RA-FLS细胞）的影响，并从抗凋亡和抗炎角度揭示其潜在分子机制。
用TNF-α刺激后，采用CCK-8法检测RA-FLS细胞的增殖情况。采用TUNEL染色检测细胞凋亡。采用蛋白质免疫印迹法检测Iκκβ、磷酸化-p65、p65、磷酸化-IκB、IκB、裂解型半胱天冬酶-3、半胱天冬酶-3、Bcl-2和Bax的表达。通过逆转录聚合酶链反应（RT-PCR）检测血红素加氧酶-1（HO-1）。采用酶联免疫吸附测定法（ELISA）测定促炎细胞因子白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、白细胞介素-8（IL-8）和基质金属蛋白酶-（MMP-1）的含量。
Galu以剂量依赖方式显著抑制细胞增殖。此外，Galu明显促进RA-FLS细胞的凋亡率，提高HO-1、半胱天冬酶-3和Bax的表达水平，同时降低Bcl-2的表达水平。此外，Galu明显抑制Iκκβ、磷酸化-p65和磷酸化-IκB的水平。此外，Galu还显著降低RA-FLS细胞中促炎因子IL-1β、IL-6、IL-8和MMP-1的水平。
木犀草素通过抑制凋亡和炎症对TNF-α诱导的RA-FLS细胞发挥保护作用，可为类风湿关节炎的临床应用提供指导。
