摘要
目的:用细胞学方法探讨桃仁膝康丸治疗骨性关节炎的作用和机制。方法:人滑膜成纤维细胞采用肿瘤坏死因子(Tumor Necrosis Factor,TNF)-α诱导建立急性炎症模型,采用脂多糖诱导建立巨噬细胞M1极化模型,人软骨细胞采用CoCl_(2)诱导建立缺氧凋亡模型,通过CCK-8法检测桃仁膝康丸对细胞的毒性,Western Blot检测全长基质金属蛋白酶9(Full-length Matrix Metalloproteinase-9,Full MMP9)、裂解基质金属蛋白酶9(Cleaved Matrix Metalloproteinase-9,Cleaved MMP9)、TNF-α、白细胞介素(Interleukin,IL)-6、全长半胱氨酸蛋白酶3(Full Caspase-3)和裂解半胱氨酸蛋白酶3(Cleaved Caspase-3)的表达。结果:1μL/mL及以下浓度的桃仁膝康丸不影响细胞活力;Western Blot结果显示,桃仁膝康丸显著抑制滑膜成纤维细胞MMP9生成、活化,以及M1巨噬细胞炎症因子生成和软骨细胞Caspase-3活化。结论:桃仁膝康丸可通过抑制基质降解酶的生成与活化、炎症因子的生成及软骨细胞凋亡发挥治疗骨性关节炎功能,其机制涉及软骨基质降解、炎症生物学过程和细胞凋亡通路,验证了网络药理学分析结果。
Objective:The effect and mechanism of Taoren Xikang Wan(桃仁膝康丸)in the treatment of osteoarthritis by cytological methods were explored.Methods:Human synovial fibroblasts were induced by TNF-αto establish an acute inflammatory model,a M1 polarization model of human macrophages were induced to established by LPS,and human chondrocytes were induced by CoCl_(2)to establish a hypoxic apoptosis model.Detection of cell toxicity of Taoren Xikang Wan was performed by CCK-8 method,and theO expressions of Full MMP9,Cleaved MMP9,TNF-α,IL-6,Full Caspase-3 and Cleaved Caspase-3 were detected by Western Blot.Results:Taoren Xikang Wan concentration of 1μL/mL and below had no effect on the viability of cells.Western Blot results showed that Taoren Xikang Wan significantly inhibited the generation and excitation of MMP9 in synovial fibroblasts,the generation of inflammatory factors in M1 polarization of macrophages,and the excitation of Caspase-3 in chondrocytes.Conclusion:Taoren Xikang Wan show therapeutic effects on osteoarthritis by inhibiting the generation and excitation of degrading enzyme,generation of inflammatory cytokines and chondrocyte apoptosis.Its mechanism involves degradation of cartilage matrix,inflammatory biological process and cell apoptosis pathway,which verifies the results of network pharmacology analysis.
出处
《中医临床研究》
2024年第7期71-76,共6页
Clinical Journal Of Chinese Medicine
基金
河南省中医药科学研究专项重点课题(2019ZY1033)。
