Department of Pharmacy, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, 325000, China.
Curr Med Sci. 2022 Oct;42(5):1046-1054. doi: 10.1007/s11596-022-2627-z. Epub 2022 Sep 30.
To explore the role and underlying mechanism of GW842166X on osteoarthritis and osteoarthritis-associated abnormal catabolism.
The extracted mouse chondrocytes were treated with GW842166X followed by lipopolysaccharide (LPS). The chondrocytes were divided into the control group, LPS group, LPS+50 nmol/L GW842166X group, and LPS+100 nmol/L GW842166X group. The cytotoxicity of GW842166X was tested using the CCK-8 assay. Western blot, RT-qPCR, and ELISA were applied to evaluate the expression of the inflammatory biomarkers in mouse chondrocytes. The expression of extracellular matrix molecules was detected by the Western blot, RT-qPCR, and immunofluorescence. Additionally, the activity of NF-κB was checked by the Western blot and immunofluorescence. The mouse Hulth models were generated to examine the in vivo effects of GW842166X on osteoarthritis. Hematoxylin and eosin staining, safranin O/fast green staining, and immunohistochemistry were applied to detect the histological changes.
GW842166X below 200 µmol/L had no cytotoxicity on the mouse chondrocytes. LPS-induced high expression of TGF-β1, IL-10, TNF-α, and IL-6 was significantly reduced by GW842166X. In addition, GW842166X upregulated the expression of aggrecan and collagen type III, which was downregulated after the LPS stimulation. The upregulated expression of ADAMTS-5 and MMP-13 by LPS stimulation was dropped in response to the GW842166X treatment. Furthermore, LPS decreased the IκBα expression in the cytoplasm and increased the nuclear p65 expression. However, these changes were reversed by the GW842166X pretreatment. Moreover, the damages in the knees caused by the Hulth surgery in mice were restored by GW842166X.
GW842166X impeded the LPS-mediated catabolism in mouse chondrocytes, thereby inhibiting the progression of osteoarthritis.
探讨 GW842166X 对骨关节炎及骨关节炎相关异常分解代谢的作用及机制。
提取小鼠软骨细胞，用 GW842166X 处理后加入脂多糖（LPS）。将软骨细胞分为对照组、LPS 组、LPS+50nmol/L GW842166X 组和 LPS+100nmol/L GW842166X 组。采用 CCK-8 法检测 GW842166X 的细胞毒性。采用 Western blot、RT-qPCR 和 ELISA 检测小鼠软骨细胞中炎症生物标志物的表达。采用 Western blot、RT-qPCR 和免疫荧光法检测细胞外基质分子的表达。采用 Western blot 和免疫荧光法检测 NF-κB 的活性。采用 Hulth 小鼠模型检测 GW842166X 对骨关节炎的体内作用。采用苏木精和伊红染色、番红 O/快绿染色和免疫组化检测组织学变化。
GW842166X 浓度低于 200μmol/L 时对小鼠软骨细胞无细胞毒性。GW842166X 可显著降低 LPS 诱导的 TGF-β1、IL-10、TNF-α和 IL-6 的高表达。此外，GW842166X 上调了 aggrecan 和胶原 III 的表达，而 LPS 刺激后其表达下调。GW842166X 处理可降低 LPS 刺激引起的 ADAMTS-5 和 MMP-13 表达上调。此外，LPS 降低了细胞质中 IκBα的表达，增加了核内 p65 的表达。然而，这些变化在 GW842166X 预处理后得到逆转。此外，GW842166X 可恢复 Hulth 手术引起的小鼠膝关节损伤。
GW842166X 抑制了 LPS 介导的小鼠软骨细胞分解代谢，从而抑制了骨关节炎的进展。
