Hong Gang
Department of Orthopedics, Beijing Chaoyang Hospital, Capital Medical University, Beijing, China.
Arch Med Sci. 2020 Feb 24;20(3):947-961. doi: 10.5114/aoms.2020.93211. eCollection 2024.
Osteoarthritis (OA) is an inflammatory disorder of synovial joints which is mainly treated with therapeutic agents showing side effects associated with the gastrointestinal (GI) and metabolic system. Consequently, there is urgent need for a potent, safe and novel agent for treating OA and related disorders. Enoxolone is a pentacyclic triterpenoid obtained from the herb liquorice. Based on earlier findings, we postulated that enoxolone may produce chondroprotective activity by exerting anti-inflammatory, anti-catabolic and oxidative stress-decreasing effects.
The chondrocytes were extracted from the femoral head articular cartilage of healthy rats. Immunofluorescence staining was done for identification of chondrocytes. Cell viability and proliferation studies were done using Cell Counting Kit-8. Apoptotic cells were identified by TUNEL assay and flow cytometry analysis. Autophagy was assessed by monodansylcadaverine assay. Western blot analysis was done for expression of proteins.
In the present study we investigated the protective effect of enoxolone on interleukin 1β (IL-1β) treated I chondrocytes . Treatment with IL-1β resulted in a significant reduction in cell viability of cells in increasing dose and time. Treatment with enoxolone along with IL-1β caused a significant decrease in growth inhibition. Also, enoxolone inhibited the IL-1β mediated apoptosis and activation of caspase-3 in cells. We also observed that enoxolone elevated the levels of p-ERK1/2, light chain 3 (LC3)-II and Beclin-1 (autophagy markers) in chondrocytes. The expression of (LC3)-II and Beclin-1 was decreased when the cells were treated with U0126 (ERK1/2 inhibitor).
Our findings demonstrate that enoxolone could suppress inflammatory signaling and apoptosis via the ERK1/2 pathway in chondrocytes.
骨关节炎（OA）是一种滑膜关节的炎症性疾病，主要使用的治疗药物会产生与胃肠道（GI）和代谢系统相关的副作用。因此，迫切需要一种有效、安全且新颖的药物来治疗OA及相关疾病。甘草次酸是从草药甘草中提取的五环三萜类化合物。基于早期研究结果，我们推测甘草次酸可能通过发挥抗炎、抗分解代谢和降低氧化应激的作用而产生软骨保护活性。
从健康大鼠的股骨头关节软骨中提取软骨细胞。通过免疫荧光染色鉴定软骨细胞。使用细胞计数试剂盒-8进行细胞活力和增殖研究。通过TUNEL检测和流式细胞术分析鉴定凋亡细胞。通过单丹磺酰尸胺检测评估自噬。通过蛋白质印迹分析检测蛋白质表达。
在本研究中，我们研究了甘草次酸对白细胞介素1β（IL-1β）处理的软骨细胞的保护作用。用IL-1β处理导致细胞活力随着剂量和时间的增加而显著降低。用甘草次酸和IL-1β共同处理导致生长抑制显著降低。此外，甘草次酸抑制细胞中IL-1β介导的凋亡和半胱天冬酶-3的激活。我们还观察到甘草次酸提高了软骨细胞中p-ERK1/2、轻链3（LC3）-II和Beclin-1（自噬标志物）的水平。当用U0126（ERK1/2抑制剂）处理细胞时，LC3-II和Beclin-1的表达降低。
我们的研究结果表明，甘草次酸可通过软骨细胞中的ERK1/2途径抑制炎症信号传导和凋亡。
