槲皮素靶向CCR1、CXCR4促进人骨髓间充质干细胞的迁移  

Quercetin targets CCR1 and CXCR4 to promote migration of human bone marrow mesenchymal stem cells

作　　者：陈双 席志鹏[1] 王楠 房晓阳 刘鑫 康然[1] 谢林[1] Chen Shuang;Xi Zhipeng;Wang Nan;Fang Xiaoyang;Liu Xin;Kang Ran;Xie Lin(Affiliated Hospital of Integrated Traditional Chinese and Western Medicine,Nanjing University of Chinese Medicine,Nanjing 210028,Jiangsu Province,China)

机构地区：[1]南京中医药大学附属中西医结合医院,江苏省南京市210028

出　　处：《中国组织工程研究》2024年第31期4945-4950,共6页Chinese Journal of Tissue Engineering Research

基　　金：江苏省干部保健科研课题(BJ19030),项目负责人:席志鹏;江苏省基础研究计划自然科学基金项目(BK20221420),项目负责人:谢林。

摘　　要：背景:槲皮素在骨髓间充质干细胞增殖和分化中起重要作用,但关于其促进骨髓间充质干细胞迁移的机制研究较少。目的:通过体外实验研究槲皮素对人骨髓间充质干细胞迁移的影响,并探讨CCR1、CXCR4的调控作用。方法:选取人骨髓间充质干细胞作为实验对象。CCK-8法检测槲皮素对人骨髓间充质干细胞增殖活性的影响;细胞划痕实验和Transwell实验分别检测槲皮素处理后人骨髓间充质干细胞的体外侵袭和迁移能力;借助分子对接技术进一步展示槲皮素与CCR1、CXCR4之间的作用关系;Western blot和实时荧光定量PCR检测槲皮素处理后迁移相关趋化因子的表达。结果与结论:①5,10μmol/L槲皮素具有显著促进人骨髓间充质干细胞增殖的作用,且药物浓度在10μmol/L时,细胞增殖效率最高;②为更好地探索槲皮素影响人骨髓间充质干细胞迁移的量效关系,选取5,10μmol/L槲皮素进行后续实验,采用川芎嗪作为阳性对照药物,实验分为空白对照组、5μmol/L槲皮素组、10μmol/L槲皮素组和100μmol/L川芎嗪组;③槲皮素处理后人骨髓间充质干细胞的体外迁移和侵袭能力呈浓度依赖性提升,10μmol/L槲皮素组的迁移效果优于川芎嗪组;④分子对接结果提示槲皮素与CCR1、CXCR4之间存在强烈的相互作用;⑤槲皮素可以上调CCR1、CXCR4蛋白和mRNA的表达;⑥该研究在细胞水平上证实了槲皮素可以通过靶向CCR1、CXCR4促进人骨髓间充质干细胞的迁移。BACKGROUND:Quercetin plays an important role in the proliferation and differentiation of bone marrow mesenchymal stem cells,but less research has been done on its mechanism of promoting the migration of bone marrow mesenchymal stem cells.OBJECTIVE:To study the effect of quercetin on the migration of human bone marrow mesenchymal stem cells through in vitro experiments,and to explore the regulatory role of CCR1 and CXCR4.METHODS:Human bone marrow mesenchymal stem cells were selected as experimental subjects.CCK8 assay was used to detect the effect of quercetin on the proliferative activity of human bone marrow mesenchymal stem cells.Cell scratch assay and Transwell assay were used to detect the in vitro invasive and migratory abilities of human bone marrow mesenchymal stem cells after quercetin treatment,respectively.The role of quercetin in relation to CCR1 and CXCR4 was demonstrated with the help of molecular docking technology.Western blot assay and real-time fluorescence quantitative PCR were used to detect the migration-related chemokine expression after quercetin treatment.RESULTS AND CONCLUSION:(1)5 and 10μmol/L quercetin could significantly promote the proliferation of human bone marrow mesenchymal stem cells,and the drug concentration of 10μmol/L resulted in the highest cell proliferation efficiency.(2)To better explore the dose-effect relationship of quercetin affecting the migration of human bone marrow mesenchymal stem cells,5 and 10μmol/L quercetin were selected for the subsequent experiments,and ligustrazine was used as the positive control drug,and the experiments were divided into blank control group,5μmol/L quercetin group,10μmol/L quercetin group,and 100μmol/L ligustrazine group.(3)In vitro migration and invasion ability of human bone marrow mesenchymal stem cells were elevated in a concentration-dependent manner after quercetin treatment,and the migration effect of 10μmol/L quercetin group was better than that of ligustrazine group.(4)The molecular docking results suggested that there was

关 键 词：骨髓间充质干细胞 槲皮素 迁移 增殖 分子对接 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R285