Xu Xiaotong, Li Naping, Wu Yongrong, Yan Ke, Mi Yilin, Yi Nanxing, Tan Xuyi, Kuang Gaoyan, Lu Min
Department of Orthopedic Surgery, The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan, China.
Graduate School, Hunan University of Chinese Medicine, Changsha, Hunan, China.
Front Pharmacol. 2022 Sep 15;13:951860. doi: 10.3389/fphar.2022.951860. eCollection 2022.
Knee osteoarthritis (KOA), a chronic degenerative disease, is mainly characterized by destruction of articular cartilage and inflammatory reactions. At present, there is a lack of economical and effective clinical treatment. Zhuifeng Tougu (ZFTG) capsules have been clinically approved for treatment of OA as they relieve joint pain and inflammatory manifestations. However, the mechanism of ZFTG in KOA remains unknown. This study aimed to investigate the effect of ZFTG on the TLR4/MyD88/NF-κB signaling pathway and its therapeutic effect on rabbits with KOA. , we established a rabbit KOA model using the modified Videman method. , we treated chondrocytes with IL-1β to induce a pro-inflammatory phenotype and then intervened with different concentrations of ZFTG. Levels of IL-1β, IL-6, TNF-α, and IFN-γ were assessed with histological observations and ELISA data. The effect of ZFTG on the viability of chondrocytes was detected using a Cell Counting Kit-8 and flow cytometry. The protein and mRNA expressions of TLR2, TLR4, MyD88, and NF-κB were detected using Western blot and RT-qPCR and immunofluorescence observation of NF-κB p65 protein expression, respectively, to investigate the mechanism of ZFTG in inhibiting inflammatory injury of rabbit articular chondrocytes and alleviating cartilage degeneration. The TLR4/MyD88/NF-κB signaling pathway in rabbits with KOA was inhibited, and the levels of IL-1β, IL-6, TNF-α, and IFN-γ in blood and cell were significantly downregulated, consistent with histological results. Both the protein and mRNA expressions of TLR2, TLR4, MyD88, NF-κB, and NF-κB p65 proteins in that nucleus decreased in the ZFTG groups. Moreover, ZFTG promotes the survival of chondrocytes and inhibits the apoptosis of inflammatory chondrocytes. ZFTG alleviates the degeneration of rabbit knee joint cartilage, inhibits the apoptosis of inflammatory chondrocytes, and promotes the survival of chondrocytes. The underlying mechanism may be inhibition of the TLR4/MyD88/NF-kB signaling pathway and secretion of inflammatory factors.
膝骨关节炎（KOA）是一种慢性退行性疾病，主要特征为关节软骨破坏和炎症反应。目前，缺乏经济有效的临床治疗方法。追风透骨（ZFTG）胶囊已获批用于骨关节炎的临床治疗，因其可缓解关节疼痛和炎症表现。然而，ZFTG治疗KOA的机制尚不清楚。本研究旨在探讨ZFTG对TLR4/MyD88/NF-κB信号通路的影响及其对KOA家兔的治疗作用。我们采用改良的Videman方法建立了家兔KOA模型。我们用IL-1β处理软骨细胞以诱导促炎表型，然后用不同浓度的ZFTG进行干预。通过组织学观察和ELISA数据评估IL-1β、IL-6、TNF-α和IFN-γ的水平。使用细胞计数试剂盒-8和流式细胞术检测ZFTG对软骨细胞活力的影响。分别采用蛋白质印迹法、RT-qPCR以及NF-κB p65蛋白表达的免疫荧光观察法检测TLR2、TLR4、MyD88和NF-κB的蛋白质和mRNA表达，以研究ZFTG抑制家兔关节软骨细胞炎症损伤和减轻软骨退变的机制。KOA家兔的TLR4/MyD88/NF-κB信号通路受到抑制，血液和细胞中IL-1β、IL-6、TNF-α和IFN-γ的水平显著下调，与组织学结果一致。ZFTG组中TLR2、TLR4、MyD88、NF-κB以及细胞核中NF-κB p65蛋白的蛋白质和mRNA表达均降低。此外，ZFTG促进软骨细胞存活并抑制炎性软骨细胞凋亡。ZFTG减轻家兔膝关节软骨退变，抑制炎性软骨细胞凋亡，促进软骨细胞存活。其潜在机制可能是抑制TLR4/MyD88/NF-κB信号通路和炎性因子分泌。
