Zhou Lei, Sun Cong Cong, Shen Fa Juan, Yuan Yu Feng, Sun Qing Lin, Rui Ze
Department of Hand Surgery, The Second Affiliated Hospital of Shandong First Medical University, Tai'an City, China.
Department of Gastroenterology, The Second Affiliated Hospital of Shandong First Medical University, Tai'an City, China.
J Biochem Mol Toxicol. 2025 Aug;39(8):e70407. doi: 10.1002/jbt.70407.
The objective of this study was to investigate whether pterostilbene ameliorates chondrocyte injury in osteoarthritis by regulating the rat sarcoma virus/rapidly accelerated fibrosarcoma/mitogen-activated protein kinase kinase/extracellular signal-regulated kinase pathway. The animal model of osteoarthritis was established, and interleukin-1β-treated chondrocytes were cultured. In vivo and in vitro models were treated with different doses of pterostilbene. Hematoxylin-eosin staining and safranin O-fast green staining were used to evaluate the pathological injury degree of cartilage tissue. Chondrocyte viability was assessed by cell counting kit-8 assay, apoptosis was detected by flow cytometry, and autophagosomes were measured by monodansylcadaverine staining. Enzyme-linked immunosorbent assay measured tumor necrosis factor-α, interleukin-1β, and interleukin-6 in cartilage tissue and chondrocytes. Western blot or real-time reverse transcriptase-polymerase chain reaction was conducted to detect matrix metalloproteinase-1, matrix metalloproteinase-13, cleaved caspase-3, phosphorylated p65, p65, p62, autophagy-related gene 5, phosphorylated extracellular signal-regulated kinase, and phosphorylated mitogen-activated protein kinase kinase in chondrocytes and tissues. Pterostilbene attenuated articular cartilage injury, repaired cartilage tissue morphology, and suppressed inflammatory factors in osteoarthritis mice. Pterostilbene reduced interleukin-1β-induced chondrocyte injury, increased cell viability, reduced apoptosis rate, promoted autophagosome formation, and inhibited levels of chondrocyte inflammatory factors. In osteoarthritis mice and interleukin-1β-treated chondrocytes, pterostilbene inhibited matrix metalloproteinase-1, matrix metalloproteinase-13, cleaved caspase-3, phosphorylated p65, and p62 expressions, and promoted autophagy-related gene 5 expression. Pterostilbene blocked rat sarcoma virus/rapidly accelerated fibrosarcoma/mitogen-activated protein kinase kinase/extracellular signal-regulated kinase pathway activation. Pterostilbene and extracellular signal-regulated kinase inhibitors had a synergistic effect in the treatment of chondrocyte injury, and extracellular signal-regulated kinase agonists reversed the therapeutic effect of pterostilbene. Pterostilbene improves chondrocyte injury by blocking rat sarcoma virus/rapidly accelerated fibrosarcoma/mitogen-activated protein kinase kinase/extracellular signal-regulated kinase pathway activation.
本研究的目的是探讨紫檀芪是否通过调节大鼠肉瘤病毒/快速加速纤维肉瘤/丝裂原活化蛋白激酶激酶/细胞外信号调节激酶通路来改善骨关节炎中的软骨细胞损伤。建立骨关节炎动物模型，并培养白细胞介素-1β处理的软骨细胞。体内和体外模型用不同剂量的紫檀芪处理。苏木精-伊红染色和番红O-固绿染色用于评估软骨组织的病理损伤程度。通过细胞计数试剂盒-8法评估软骨细胞活力，通过流式细胞术检测细胞凋亡，通过单丹磺酰尸胺染色测量自噬体。酶联免疫吸附测定法检测软骨组织和软骨细胞中的肿瘤坏死因子-α、白细胞介素-1β和白细胞介素-6。进行蛋白质免疫印迹或实时逆转录-聚合酶链反应以检测软骨细胞和组织中的基质金属蛋白酶-1、基质金属蛋白酶-13、裂解的半胱天冬酶-3、磷酸化的p65、p65、p62、自噬相关基因5、磷酸化的细胞外信号调节激酶和磷酸化的丝裂原活化蛋白激酶激酶。紫檀芪减轻了骨关节炎小鼠的关节软骨损伤，修复了软骨组织形态，并抑制了炎症因子。紫檀芪减少了白细胞介素-1β诱导的软骨细胞损伤，提高了细胞活力，降低了凋亡率，促进了自噬体形成，并抑制了软骨细胞炎症因子水平。在骨关节炎小鼠和白细胞介素-1β处理的软骨细胞中，紫檀芪抑制基质金属蛋白酶-1、基质金属蛋白酶-13、裂解的半胱天冬酶-3、磷酸化的p65和p62表达，并促进自噬相关基因5表达。紫檀芪阻断了大鼠肉瘤病毒/快速加速纤维肉瘤/丝裂原活化蛋白激酶激酶/细胞外信号调节激酶通路的激活。紫檀芪与细胞外信号调节激酶抑制剂在治疗软骨细胞损伤方面具有协同作用，而细胞外信号调节激酶激动剂则逆转了紫檀芪的治疗效果。紫檀芪通过阻断大鼠肉瘤病毒/快速加速纤维肉瘤/丝裂原活化蛋白激酶激酶/细胞外信号调节激酶通路的激活来改善软骨细胞损伤。
