Department of orthopedics, Traditional Chinese Medicine Hospital of Xiaoshan District, Zhejiang, China.
Department of orthopedics, Jiangnan Hospital Affiliated to Zhejiang Chinese Medical University, Zhejiang, China.
Orthop Surg. 2020 Aug;12(4):1319-1326. doi: 10.1111/os.12745. Epub 2020 Jul 23.
Wufu Decoction (WFD) is a herbal formulation composed of five traditional Chinese herbs that is used clinically for arthritis treatment in China. The current study investigated the chondroprotective effects and the underlying mechanism of WFD for osteoarthritis (OA) therapy.
The chondroprotective effects of WFD were investigated based on vitro study. Following the successful isolation of chondrocytes from rat cartilage tissues and the identification of collagen II expression with immunofluorescence staining, chondrocytes were co-incubated with tumor necrosis factor-α (TNF-α) to induce an inflammation model; WFD was also administered. After the treatment, cell viability was determined by MTT assay, cell apoptosis was assessed by DAPI staining, the concentration of inflammation cytokines interleukin (IL)-1β and IL-6 were detected with ELISA assay, the expression of collagen II, MEK1/2-ERK1/2 signaling pathway proteins was detected using western blotting, and mRNA expression of MMP-1, MMP-9 and MMP-13 were determined with quantitative real-time polymerase chain reaction.
Wufu Decoction significantly restored the cell viability suppressed by TNF-α and inhibited the cell apoptosis induced by TNF-α in chondrocytes. The high concentrations of IL-1β and IL-6 in TNF-α-induced model cells were significantly decreased in WFD-treated chondrocytes, and the immunofluorescence staining and western blot results showed that the inhibited expression of collagen II in the TNF-α-induced model group was significantly increased in WFD-treated chondrocytes. The protein expressions of MEK1/2, p-ERK1/2, and P53 were significantly reduced in the WFD-treated group compared with those in the model group, and the mRNA expressions of MMP-1, MMP-9, and MMP-13 were also significantly reduced with WFD treatment.
The present study indicated that WFD exerted a chondroprotective effect in TNF-α-induced chondrocytes via the regulation of the ERK1/2 signaling pathway, suggesting that WFD has therapeutic potential for OA therapy.
五辐散（WFD）是一种由五种中草药组成的中药配方，临床上用于治疗中国的关节炎。本研究探讨了 WFD 对骨关节炎（OA）治疗的软骨保护作用及其潜在机制。
在体外研究中考察了 WFD 的软骨保护作用。成功从大鼠软骨组织中分离出软骨细胞，并通过免疫荧光染色鉴定 II 型胶原的表达后，将软骨细胞与肿瘤坏死因子-α（TNF-α）共孵育，诱导炎症模型；同时给予 WFD 治疗。治疗后，通过 MTT 法测定细胞活力，DAPI 染色法评估细胞凋亡，ELISA 法检测炎症细胞因子白细胞介素（IL）-1β和 IL-6 的浓度，western blot 法检测胶原 II、MEK1/2-ERK1/2 信号通路蛋白的表达，定量实时聚合酶链反应法测定 MMP-1、MMP-9 和 MMP-13 的 mRNA 表达。
WFD 显著恢复了 TNF-α抑制的细胞活力，并抑制了 TNF-α诱导的软骨细胞凋亡。TNF-α诱导模型细胞中高浓度的 IL-1β和 IL-6在 WFD 处理的软骨细胞中显著降低，免疫荧光染色和 western blot 结果显示，TNF-α诱导模型组中抑制表达的胶原 II 在 WFD 处理的软骨细胞中显著增加。与模型组相比，WFD 处理组中 MEK1/2、p-ERK1/2 和 P53 的蛋白表达显著降低，WFD 处理后 MMP-1、MMP-9 和 MMP-13 的 mRNA 表达也显著降低。
本研究表明，WFD 通过调节 ERK1/2 信号通路对 TNF-α诱导的软骨细胞发挥软骨保护作用，提示 WFD 具有治疗 OA 的潜力。
