Pirozzi Claudio, Francisco Vera, Guida Francesca Di, Gómez Rodolfo, Lago Francisca, Pino Jesus, Meli Rosaria, Gualillo Oreste
SERGAS (Servizo Galego de Saude) and IDIS (Instituto de Investigación Sanitaria de Santiago), The NEIRID Lab (Neuroendocrine Interactions in Rheumatology and Inflammatory Diseases), Research Laboratory 9, Santiago University Clinical Hospital, Santiago de Compostela,
Department of Pharmacy. University of Naples Federico II, Naples,
Cell Physiol Biochem. 2018;51(1):228-243. doi: 10.1159/000495203. Epub 2018 Nov 16.
BACKGROUND/AIMS: Osteoarthritis (OA) is a joint degenerative biomechanical disorder involving immunity, metabolic alterations, inflammation, and cartilage degradation, where chondrocytes play a pivotal role. OA has not effective pharmacological treatments and new therapeutic targets are needed. Adipokines contribute to the low-grade systemic inflammation in OA. Here, we explored novel molecular mechanisms of sodium butyrate (BuNa) in modulating inflammation and chemotaxis in chondrocytes, demonstrating the direct involvement of its G protein-coupled receptor (GPR)-43.
ATDC5 murine chondrocytes were stimulated with interleukin (IL)-1β, in the presence or not of BuNa, for 24 h. RT-PCR and Western blot analysis was performed to evaluate the expression of inflammatory mediators and structural proteins.
Butyrate reduced the expression of canonic pro-inflammatory mediators (Nos2, COX-2, IL-6), pro-inflammatory adipokines (lipocalin-2 and nesfatin-1) and adhesion molecule (VCAM-1 and ICAM-1) in IL-1β-stimulated chondrocytes, inhibiting several inflammatory signalling pathways (NFκB, MAPKinase, AMPK-α, PI3K/Akt). Butyrate also reduced metalloproteinase production and limited the loss of type II collagen in IL-1β-inflamed chondrocytes. The chemoattractant effect of butyrate, after different inflammatory challenges, was revealed by increased annexin (AnxA)1 levels and chemokines expression. The chemoattractant and anti-inflammatory activities of butyrate were completely blunted by GPR43 silencing using RNA interference.
Taken together, our data suggest the potential application of sodium butyrate as a novel candidate in a multi-target approach for the treatment of chondrocyte inflammation and cartilage degenerative process.
背景/目的:骨关节炎(OA)是一种涉及免疫、代谢改变、炎症和软骨降解的关节退行性生物力学疾病,其中软骨细胞起关键作用。OA尚无有效的药物治疗方法,需要新的治疗靶点。脂肪因子促成了OA中的低度全身炎症。在此,我们探究了丁酸钠(BuNa)调节软骨细胞炎症和趋化性的新分子机制,证明了其G蛋白偶联受体(GPR)-43的直接参与。
在有或无BuNa的情况下,用白细胞介素(IL)-1β刺激ATDC5小鼠软骨细胞24小时。进行逆转录-聚合酶链反应(RT-PCR)和蛋白质免疫印迹分析以评估炎症介质和结构蛋白的表达。
丁酸盐降低了IL-1β刺激的软骨细胞中典型促炎介质(Nos2、COX-2、IL-6)、促炎脂肪因子(lipocalin-2和nesfatin-1)和黏附分子(VCAM-1和ICAM-1)的表达,抑制了几种炎症信号通路(NFκB、丝裂原活化蛋白激酶、AMPK-α、磷脂酰肌醇-3激酶/蛋白激酶B)。丁酸盐还减少了金属蛋白酶的产生,并限制了IL-1β炎症刺激的软骨细胞中II型胶原蛋白的损失。不同炎症刺激后,丁酸盐的趋化作用通过膜联蛋白(AnxA)1水平和趋化因子表达增加得以体现。使用RNA干扰使GPR43沉默后,丁酸盐的趋化和抗炎活性完全丧失。
综上所述,我们的数据表明丁酸钠作为一种新型候选药物,在多靶点治疗软骨细胞炎症和软骨退变过程中具有潜在应用价值。