摘要
Objective: This study aims to clarify the effect of the active components puerarin and tetrandrine on the chondrogenic differentiation of bone marrow mesenchymal stem cells(BMSCs).Methods: Using network pharmacology, protein targets of puerarin and tetrandrine were predicted, and a database of cartilage formation targets was established. The protein target information related to disease was then collected, and the drug-targeting network was constructed by analyzing the protein–protein interactions. Genes related to chondrogenesis induced by puerarin and tetrandrine and chondroblast differentiation signaling pathways were searched. Finally, potential drug-and disease-related genes,as well as proteins, were screened and verified using real-time RT-PCR and western blotting.Results: Network pharmacological studies have shown that puerarin and tetrandrine are involved in BMSCs cartilage differentiation. The experimental results showed that puerarin and tetrandrine could regulate the expression of cartilage differentiation-related genes and proteins. Puerarin increased the protein expression of COL2 A1, COL10 A1, MMP13, and SOX-9,as well as the gene expression of Col2 a1, Mmp13, Tgfb1, and Sox-9. Tetrandrine increased the protein expression of COL2 A1,COL10 A1, MMP13, and SOX-9, as well as the gene expression of Col10 a1, Tgfb1, Sox-9, and Acan. The combination of puerarin and tetrandrine increased the protein expression of COL2 A1, COL10 A1, MMP13, and SOX-9 and the gene expression of Col2 a1,Col10 a1, Sox-9, and Acan.Conclusions: Puerarin, tetrandrine, and their combination can promote the proliferation of BMSCs and induce their differentiation into chondrocytes, and they are thus expected to be inducers of chondrogenic differentiation. These results suggest that puerarin and tetrandrine have potential therapeutic effects on osteoarthritis.
目的:本研究旨在阐明中药活性成分葛根素和粉防己碱对骨髓间充质干细胞向软骨分化的影响,从而探究其对骨关节炎的保护作用。方法:首先,通过网络药理学手段,预测葛根素和粉防己碱的作用靶点并建立软骨形成靶点数据库;其次,收集与活性成分相关的靶蛋白的靶点信息,通过分析蛋白与蛋白间的相互作用构建药物靶向网络;然后,筛选葛根素和粉防己碱与软骨形成的相关基因,以及与成软骨细胞分化的相关信号通路;最后,运用Real-Time RT-PCR和Western blot技术,对潜在的药物和疾病相关基因以及蛋白质的表达进行验证。结果:网络药理学研究结果表明,葛根素和粉防己碱与骨髓间充质干细胞的软骨分化有关。实验结果表明,葛根素和粉防己碱对软骨分化相关蛋白和基因的表达具有调控作用。葛根素可以上调COL2A1、COL10A1、MMP13、SOX-9蛋白以及Col2a1、Mmp13、Tgfb1、Sox-9基因的表达;粉防己碱可以上调COL2A1、COL10A1、MMP13、SOX-9蛋白以及Col10a1、Tgfb1、Sox-9、Acan基因的表达;葛根素与粉防己碱的联合用药可以上调COL2A1、COL10A1、MMP13、SOX-9蛋白以及Col2a1、Col10a1、Sox-9、Acan基因的表达。结论:葛根素、粉防己碱及其联合用药可以促进骨髓间充质干细胞增殖并诱导其分化为软骨细胞,有望成为软骨分化的诱导剂。以上结果表明,葛根素和粉防己碱对骨关节炎具有潜在的治疗作用。
基金
supported by the NSFC(No.81473542)。
