白藜芦醇通过上调SIRT1/RUNX2表达促进多发性骨髓瘤病人骨髓间充质干细胞成骨分化  

Resveratrol Promotes Osteogenic Differentiation of Multiple Myeloma Derived Bone Marrow Mesenchymal Stem Cells Via Upregulating SIRT1/RUNX2

作　　者：潘杰 陶叠宏[2] 任莉[2] 苏传勇[2] 陈志炉[2] 蒋慧芳[2] 王珏 钱令波 PAN Jie;TAO Die-Hong;REN Li;SU Chuan-Yong;CHEN Zhi-Lu;JIANG Hui-Fang;WANG Jue;QIAN Ling-Bo(Department of Physiology,School of Basic Medical Sciences&Forensic Medicine,Hangzhou Medical College,Hangzhou 310053,China;Department of Hematology,Tongde Hospital of Zhejiang Province,Hangzhou 310012,China)

机构地区：[1]杭州医学院基础医学与法医学院生理学教研室,杭州310053 [2]浙江省立同德医院血液科,杭州310012

出　　处：《中国生物化学与分子生物学报》2021年第3期354-362,共9页Chinese Journal of Biochemistry and Molecular Biology

基　　金：浙江省教育厅一般科研项目(No.Y201840344);浙江省“十三五”第二批教学改革研究项目(No.jg20190549);浙江省第三期医坛新秀培养项目(2017);浙江省基础公益研究计划项目(No.LQY19H080001);浙江省医药卫生科技计划项目(No.2019KY049);魏克民全国名老中医药专家传承工作室资助。

摘　　要：骨髓瘤骨病(myeloma bone disease,MBD)是多发性骨髓瘤(multiple myeloma,MM)最主要的并发症之一。骨髓瘤骨病骨损伤发生的病因是肿瘤细胞浸润以及骨髓微环境改变等因素导致的破骨细胞活化和成骨细胞抑制。骨髓间充质干细胞(bone marrow mesenchymal stem cell,BMSC)是成骨细胞的主要来源,促进BMSC的成骨分化可能是治疗骨髓瘤骨病的可行方法之一。白藜芦醇(resveratrol,RES)是一种天然多酚类植物激素。已有研究发现,白藜芦醇具有调节骨质代谢的功能。但白藜芦醇对多发性骨髓瘤病人骨髓间充质干细胞(MM-BMSC)成骨分化的作用和机制尚不明确。本研究成功分离、培养并鉴定了10例MM-BMSC。通过检测碱性磷酸酶(alkaline phosphatase,ALP)活性和骨钙素(osteocalcin,OCN)水平,以及茜素红染色发现,白藜芦醇具有促进多发性骨髓瘤病人骨髓间充质干细胞成骨分化的作用。生物信息学分析提示,沉默信息调节因子1(silent information regulator 1,SIRT1)可能是白藜芦醇调控多发性骨髓瘤病人骨髓间充质干细胞成骨分化的靶基因。体外细胞研究发现,白藜芦醇可增加SIRT1表达(P<0.001),且用siRNA干扰SIRT1表达后多发性骨髓瘤病人骨髓间充质干细胞碱性磷酸酶活性下降(P<0.05),骨钙素表达减少(P<0.01),茜素红染色矿化结节减少。多发性骨髓瘤病人骨髓间充质干细胞经白藜芦醇处理后,Runt相关转录因子2(runt related transcription factor 2,RUNX2)mRNA(P<0.001)和蛋白质(P<0.01)表达水平上升。应用siRNA干扰多发性骨髓瘤病人骨髓间充质干细胞SIRT1表达后,细胞RUNX2 mRNA(P<0.01)和蛋白质(P<0.05)表达均下降。综上所述,本研究发现,白藜芦醇通过上调SIRT1/RUNX2表达促进多发性骨髓瘤病人骨髓间充质干细胞成骨分化。白藜芦醇有可能成为骨髓瘤骨病治疗的潜在有效药物。Myeloma bone disease(MBD)is one of the most common complications of multiple myeloma(MM).MBD is considered to be caused by the activation of osteoclasts and suppression of osteoblasts resulting from the involvement of neoplastic plasma cells and the change of bone marrow microenvironment.It may be a feasible way to improve the treatment of MBD by promoting osteogenic differentiation of bone marrow mesenchymal stem cell(BMSC),from which the osteoblasts mainly originate.Resveratrol(RES),a naturally occurring polyphenolic flavonoid compound,was reported to function in the modulation of bone metabolism.But the effects of RES on osteogenic differentiation of MM derived BMSC(MM-BMSC)and its underlying mechanism remains unknown.Totally 10 cases of MM-BMSCs were isolated,cultured and identified successfully in the present study.RES was found to promote osteogenic differentiation of MM-BMSC by alkaline phosphatase activity assay,qRT-PCR and alizarin red staining.SIRT1 was predicted to be the target gene of RES in promoting osteogenic differentiation with bioinformatic analysis.RES upregulated the expression of silent information regulator 1(SIRT1)in MM-BMSC(P<0.001)and its osteogenic differentiation was inhibited in the SIRT1 small interfering RNA(si-SIRT1)transfected group.Furthermore,the mRNA(P<0.001)and protein(P<0.01)expression of runt related transcription factor 2(RUNX2)was increased in the RES treated group and decreased(mRNA P<0.01,protein P<0.05)in si-SIRT1 transfected group,respectively.In conclusion,resveratrol promotes osteogenic differentiation of MM-BMSCs via upregulating SIRT1/RUNX2 and seems to be a potential therapeutic agent to counteract bone disease in MM patients.

关 键 词：白藜芦醇 多发性骨髓瘤 骨髓间充质干细胞 成骨分化 沉默信息调节因子1 

分 类 号：R559[医药卫生—血液循环系统疾病]