Department of Orthopedic Surgery, Second Clinical College, Dalian Medical University, No. 222 Zhongshan Road, Dalian, 116011, China.
Dalian love cubic Health Management Co., Ltd, Dalian, 116000, China.
Inflammation. 2019 Aug;42(4):1265-1275. doi: 10.1007/s10753-019-00986-3.
Sinomenine (SIN) has been shown to protect against IL-1β-induced chondrocyte apoptosis in vitro. However, the role of SIN in the anterior cruciate ligament transection (ACLT)-induced osteoarthritis (OA) mouse model and its underlying molecular mechanisms remain unclear. In the present study, the protective effect of SIN on ACLT-induced articular cartilage degeneration and IL-1β-induced chondrocyte apoptosis miR-223-3p/NLRP3 signaling regulation was investigated. Safranin O staining was performed to evaluate the pathological changes of articular cartilage. Chondrocyte apoptosis was measured with Annexin V-fluorescein isothiocyanate/polyimide (annexin V-FITC/PI) staining using flow cytometry. Gene and protein expression were detected by RT-qPCR and Western blotting, respectively. SIN administration markedly improved articular cartilage degradation in mice undergoing ACLT surgery. In addition, SIN treatment downregulated the levels of inflammatory cytokines and the protein expression of NLRP3 inflammasome components and upregulated the expression of miR-223-3p in OA mice and IL-1β-stimulated chondrocytes. In vitro, we found that NLRP3 was a direct target of miR-223-3p, and overexpression of miR-223-3p blocked IL-1β-induced apoptosis and the inflammatory response in chondrocytes. These findings indicate that miR-223-3p/NLRP3 signaling could be used as a potential target of SIN for the treatment of OA.
青藤碱（SIN）已被证明可在体外防止白细胞介素 1β诱导的软骨细胞凋亡。然而，SIN 在前交叉韧带切断（ACLT）诱导的骨关节炎（OA）小鼠模型中的作用及其潜在的分子机制尚不清楚。在本研究中，研究了 SIN 对 ACLT 诱导的关节软骨退化和 IL-1β诱导的软骨细胞凋亡 miR-223-3p/NLRP3 信号调节的保护作用。番红 O 染色用于评估关节软骨的病理变化。通过流式细胞术用 Annexin V-异硫氰酸荧光素/聚酰亚胺（annexin V-FITC/PI）染色测量软骨细胞凋亡。通过 RT-qPCR 和 Western blot 分别检测基因和蛋白表达。SIN 给药显著改善了接受 ACLT 手术的小鼠的关节软骨降解。此外，SIN 治疗下调了 OA 小鼠和 IL-1β 刺激的软骨细胞中炎症细胞因子和 NLRP3 炎症小体成分的蛋白表达水平，并上调了 miR-223-3p 的表达。在体外，我们发现 NLRP3 是 miR-223-3p 的直接靶标，过表达 miR-223-3p 阻断了 IL-1β 诱导的软骨细胞凋亡和炎症反应。这些发现表明 miR-223-3p/NLRP3 信号通路可作为 SIN 治疗 OA 的潜在靶点。
