脱水淫羊藿素对髓核间充质干细胞增殖和细胞外基质合成代谢的影响

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中文摘要：目的:研究脱水淫羊藿素(AHI)对髓核间充质干细胞(NPMSCs)增殖和细胞外基质(ECM)合成代谢的影响.方法:分离和培养NPMSCs,通过流式细胞术鉴定细胞表面抗原,采用CCK-8法检测AHI对NPMSCs增殖的影响,采用集落形成实验检测AHI对NPMSCs集落形成能力的影响.通过加入TNF-α诱导体外建立NPMSCs退变模型,采用实时荧光定量聚合酶链式反应(RT-PCR)检测NPMSCs中细胞周期蛋白依赖性激酶(CDK)2、CDK4和CDK6 的表达,采用Western blot检测CDK2、CDK4和CDK6的蛋白水平,观察AHI对CDK的影响及NPMSCs的增殖机制.采用RT-PCR检测NPMSCs中COL2A1、SOX9和ACAN的表达,采用Western blot检测SOX9和ACAN蛋白水平,观察AHI对ECM合成代谢的作用机制.结果:AHI促进NPMSCs增殖,增强NPMSCs集落形成的数量.AHI上调CDK2,CDK4和CDK6 mRNA的表达,且CDK2,CDK4和CDK6蛋白水平增高.同时AHI促进COL2A1、SOX9和ACAN的表达,SOX9和ACAN的蛋白水平显著增高.结论:AHI在一定浓度范围内能够通过调控CDK2、CDK4和CDK6促进NPMSCs增殖,通过调控SOX9和ACAN促进ECM合成代谢.

外文标题：Effects of anhydroicaritin on the proliferation and extracellular matrix synthesis of nucleus pulposus mesenchymal stem cells

外文摘要：Objective:To investigate the effects of anhydroicaritin(AHI)on the proliferation and extracellular matrix(ECM)synthesis of nucleus pulposus mesenchymal stem cells(NPMSCs).Methods:NPMSCs were isolated and cultured,and cell surface antigens were identified by flow cytometry.The effects of AHI on the proliferation of NPMSCs were detected by CCK-8 assay,and the colony formation assay was used to evaluate the impact of AHI on the colony-forming ability of NPMSCs.TNF-α-induced NPMSCs degeneration model was established in vitro.Reverse transcription polymerase chain reaction(RT-PCR)was used to detect the mRNA expression of cyclin-dependent kinase(CDK)2,CDK4 and CDK6 in NPMSCs,and the protein level of CDK2,CDK4,and CDK6 was detected by Western blotting to observe the effect of AHI on CDK and the mechanism of regulating NPMSCs proliferation.The mRNA expression of COL2A1,SOX9 and ACAN in NPMSCs was detected by RT-PCR,and the protein level of SOX9 and ACAN was detected by Western blotting to elucidate the mechanisms of AHI on ECM anabolism.Results:AHI significantly promoted the proliferation of NPMSCs and increased the number of NPMSCs colonies.AHI upregulated the mRNA expression of CDK2,CDK4,and CDK6,and increased the protein level of CDK2,CDK4,and CDK6.Meanwhile,AHI promoted the mRNA expression of COL2A1,SOX9 and ACAN,and significantly increased the protein level of SOX9 and ACAN.Conclusions:Within a certain concentration range,AHI can promote the proliferation of NPMSCs by regulating CDK2,CDK4,and CDK6,and promote the ECM synthesis by modulating SOX9 and ACAN.

外文关键词：

AnhydroicaritinNucleus Pulposus Mesenchymal Stem CellsProliferationExtracellular MatrixIntervertebral Disc Degeneration

作者：

崔鹏雷、绳月阳、吴成爱、何达

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作者单位：

首都医科大学附属北京积水潭医院脊柱外科,北京 100035

首都医科大学附属北京积水潭医院,北京市创伤骨科研究所,北京 100035

关键词：

脱水淫羊藿素髓核间充质干细胞增殖细胞外基质椎间盘退变

基金：

国家自然科学基金项目国家骨科与运动康复临床医学研究中心、江苏省中以产业技术研究院"以临床应用为导向的医疗创新基金"资助北京市博士后工作经费资助项目

项目编号：

822027192021-NCRC-CXJJ-ZH-352022-ZZ-020

出版年：

2024

DOI：

10.3969/j.issn.2095-9958.2024.06.10

中华骨与关节外科杂志

中国医学科学院中国协和医学院

中华骨与关节外科杂志

CSTPCD北大核心

影响因子：0.906

ISSN：2095-9958

年,卷(期)：2024.17(6)