中国组织工程研究

二甲基氧化甘氨酸对人骨髓间充质干细胞成骨、成脂分化及线粒体自噬的影响

PDF下载

陈启衡 ¹,,² , 翁土军 ¹, , 彭江 ¹,

中国组织工程研究 | 骨髓干细胞 2026,30(1): 50-57

收起

中国组织工程研究 |骨髓干细胞 2026 , 30 (1) : 50 -57

二甲基氧化甘氨酸对人骨髓间充质干细胞成骨、成脂分化及线粒体自噬的影响

全屏

[Author(id=1321500927832469947, tenantId=1146029695717560320, journalId=1246415837536497731, articleId=1321500922446983602, orderNo=1, firstName=null, middleName=null, lastName=null, nameCn=陈启衡, orcid=0009-0007-7265-335X, stid=null, country=null, authorPic=null, dead=null, email=[], emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=2, ext={CN=AuthorExt(id=1321500927928938942, tenantId=1146029695717560320, journalId=1246415837536497731, articleId=1321500922446983602, authorId=1321500927832469947, language=CN, stringName=陈启衡, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=¹,,², address=³,^2.放军医学院，北京市 100853放军总医院第四医学中心骨科医学部研究所，北京市 100853, bio=陈启衡: 陈启衡，男，1997年生，北京市人，汉族，解放军医学院在读硕士，主要从事股骨头坏死组织工程修复研究, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1321500927765361081, tenantId=1146029695717560320, journalId=1246415837536497731, articleId=1321500922446983602, xref=3^*, ext=[AuthorCompanyExt(id=1321500927769555386, tenantId=1146029695717560320, journalId=1246415837536497731, articleId=1321500922446983602, companyId=1321500927765361081, language=CN, country=null, province=null, city=null, postcode=null, companyName=放军总医院第四医学中心骨科医学部研究所，北京市 100853, departmentName=null, remark=^3.放军总医院第四医学中心骨科医学部研究所，北京市 100853)]), AuthorCompany(id=1321500927681474999, tenantId=1146029695717560320, journalId=1246415837536497731, articleId=1321500922446983602, xref=2, ext=[AuthorCompanyExt(id=1321500927689863608, tenantId=1146029695717560320, journalId=1246415837536497731, articleId=1321500922446983602, companyId=1321500927681474999, language=CN, country=null, province=null, city=null, postcode=null, companyName=放军医学院，北京市 100853, departmentName=null, remark=^2.放军医学院，北京市 100853)])]), Author(id=1321500927996047807, tenantId=1146029695717560320, journalId=1246415837536497731, articleId=1321500922446983602, orderNo=2, firstName=null, middleName=null, lastName=null, nameCn=翁土军, orcid=null, stid=null, country=null, authorPic=null, dead=null, email=null, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=2, ext={CN=AuthorExt(id=1321500928084128193, tenantId=1146029695717560320, journalId=1246415837536497731, articleId=1321500922446983602, authorId=1321500927996047807, language=CN, stringName=翁土军, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=¹,, address=³放军总医院第四医学中心骨科医学部研究所，北京市 100853, bio=翁土军: 彭江，主任医师，研究员，解放军总医院第四医学中心骨科医学部研究所，北京市 100853, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1321500927765361081, tenantId=1146029695717560320, journalId=1246415837536497731, articleId=1321500922446983602, xref=3^*, ext=[AuthorCompanyExt(id=1321500927769555386, tenantId=1146029695717560320, journalId=1246415837536497731, articleId=1321500922446983602, companyId=1321500927765361081, language=CN, country=null, province=null, city=null, postcode=null, companyName=放军总医院第四医学中心骨科医学部研究所，北京市 100853, departmentName=null, remark=^3.放军总医院第四医学中心骨科医学部研究所，北京市 100853)])]), Author(id=1321500928142848450, tenantId=1146029695717560320, journalId=1246415837536497731, articleId=1321500922446983602, orderNo=3, firstName=null, middleName=null, lastName=null, nameCn=彭江, orcid=null, stid=null, country=null, authorPic=null, dead=null, email=null, emailSecond=null, emailThird=null, correspondingAuthor=1, authorType=2, ext={CN=AuthorExt(id=1321500928222540228, tenantId=1146029695717560320, journalId=1246415837536497731, articleId=1321500922446983602, authorId=1321500928142848450, language=CN, stringName=彭江, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=¹,, address=³放军总医院第四医学中心骨科医学部研究所，北京市 100853, bio=彭江: 翁土军，副研究员，解放军总医院第四医学中心骨科医学部研究所，北京市 100853, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1321500927765361081, tenantId=1146029695717560320, journalId=1246415837536497731, articleId=1321500922446983602, xref=3^*, ext=[AuthorCompanyExt(id=1321500927769555386, tenantId=1146029695717560320, journalId=1246415837536497731, articleId=1321500922446983602, companyId=1321500927765361081, language=CN, country=null, province=null, city=null, postcode=null, companyName=放军总医院第四医学中心骨科医学部研究所，北京市 100853, departmentName=null, remark=^3.放军总医院第四医学中心骨科医学部研究所，北京市 100853)])])]

陈启衡¹,,², 翁土军¹,, 彭江¹,

作者信息

^1.放军总医院第四医学中心骨科医学部研究所，北京市 100853

^2.放军医学院，北京市 100853

^3.放军总医院第四医学中心骨科医学部研究所，北京市 100853

Effect of dimethylglyoxal glycine on osteogenic, adipogenesis differentiation, and mitophagy of human bone marrow mesenchymal stem cells

Affiliations

出版时间: 2026-01-08 doi: 10.12307/2025.578

文章导航

摘要

收起

背景：骨髓间充质干细胞在股骨头坏死等疾病的治疗中发挥重要作用，其治疗效果与细胞质量息息相关，如何为细胞赋能成为如今研究的热点。目的：探究低氧模拟剂二甲基氧化甘氨酸预处理对人骨髓间充质干细胞线粒体自噬、分化能力的影响。方法：从患者髂骨的骨髓血中提取骨髓间充质干细胞，体外培养至第3代，用0，10，50，100 μmol/L 二甲基氧化甘氨酸处理24 h，然后更换成骨诱导分化培养基，此为预处理组；而在细胞贴壁后直接用含有0，10，50，100 μmol/L二甲基氧化甘氨酸的成骨诱导培养基培养，此为持续处理组。诱导7 d后进行碱性磷酸酶染色，选出最利于成骨分化的条件为实验组，正常培养的骨髓间充质干细胞为对照组。采用碱性磷酸酶染色、碱性磷酸酶活性、油红O染色及RT-qPCR评估两组骨髓间充质干细胞的成骨、成脂分化差异；采用MitoSox染色检测线粒体活性氧水平；采用Mito-tracker和Lyso-tracker染色检测线粒体和溶酶体的共定位情况；使用荧光探针JC-1检测线粒体膜电位。结果与结论：碱性磷酸酶染色显示，对骨髓间充质干细胞成骨最有利的处理方式为10 μmol/L二甲基氧化甘氨酸预处理24 h。与对照组相比，实验组骨髓间充质干细胞的碱性磷酸酶染色表达增强，碱性磷酸酶活性和成骨相关基因表达升高，油红O染色显示脂滴生成减少以及成脂相关基因表达下降，线粒体活性氧生成减少，线粒体和溶酶体的共定位增多，线粒体膜电位升高。结果表明，10 μmol/L二甲基氧化甘氨酸预处理可以促进骨髓间充质干细胞成骨分化，抑制成脂分化，并增强线粒体自噬。

关键词

骨髓间充质干细胞 / 二甲基氧化甘氨酸 / 预处理 / 成骨分化 / 成脂分化 / 线粒体 / 线粒体自噬 / 线粒体膜电位 / 线粒体活性氧 / 溶酶体

Abstract

收起

BACKGROUND: Bone marrow mesenchymal stem cells play a crucial role in treatment of diseases, such as femoral head necrosis, and the therapeutic efficacy is closely related to the quality of the cells. The empowerment of cells has become a research focus. OBJECTIVE: To investigate the effects of hypoxia mimetic dimethylglyoxal glycine pretreatment on mitophagy and differentiation capacity of human bone marrow mesenchymal stem cells. METHODS: Bone marrow mesenchymal stem cells were extracted from the bone marrow of patients’ iliac crest and cultured in vitro to the third passage. The cells were treated with dimethylglyoxal glycine at 0, 10, 50, and 100 μmol/L for 24 hours, followed by the replacement with an osteogenic induction differentiation medium, which constituted the pretreatment group. The continuous treatment group was cultured directly in osteogenic induction medium containing 0, 10, 50, and 100 μmol/L dimethylglyoxal glycine after cell adhesion. After 7 days of induction, alkaline phosphatase staining was performed to select the most favorable conditions for osteogenic differentiation for subsequent experiments, with normally cultured bone marrow mesenchymal stem cells serving as the control group. Alkaline phosphatase staining, alkaline phosphatase activity, oil red O staining, and related RT-qPCR were used to evaluate the osteogenic and adipogenic differentiation differences of bone marrow mesenchymal stem cells between the two groups. MitoSox staining was used to detect mitochondrial reactive oxygen species levels. Mito-tracker and Lyso-tracker staining were used to detect the co-localization of mitochondria and lysosomes. The fluorescent probe JC-1 was used to measure mitochondrial membrane potential. RESULTS AND CONCLUSION: Alkaline phosphatase staining indicated that the most beneficial treatment for bone marrow mesenchymal stem cell osteogenesis was pretreatment with 10 μmol/L dimethylglyoxal glycine for 24 hours. Compared with the control group, the experimental group showed enhanced alkaline phosphatase staining expression, increased alkaline phosphatase activity and osteogenic gene expression, reduced lipid droplet formation and adipogenic gene expression as indicated by oil red O staining, decreased mitochondrial reactive oxygen species production, increased co-localization of mitochondria and lysosomes, and elevated mitochondrial membrane potential. The results suggest that 10 μmol/L dimethylglyoxal glycine pretreatment can promote osteogenic differentiation of bone marrow mesenchymal stem cells, inhibit adipogenic differentiation, and enhance mitophagy.

Key words

bone marrow mesenchymal stem cell / dimethylglyoxal glycine / pretreatment / osteogenic differentiation / adipogenic differentiation / mitochondria / mitophagy / mitochondrial membrane potential / mitochondrial reactive oxygen species / lysosome

引用本文

陈启衡, 翁土军, 彭江. 二甲基氧化甘氨酸对人骨髓间充质干细胞成骨、成脂分化及线粒体自噬的影响. 中国组织工程研究, 2026 , 30 (1) : 50 -57 . DOI: 10.12307/2025.578

. Effect of dimethylglyoxal glycine on osteogenic, adipogenesis differentiation, and mitophagy of human bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2026 , 30 (1) : 50 -57 . DOI: 10.12307/2025.578

基金

收起

国家重点研发计划课题
解放军总医院青年自主创新科学基金项目

参考文献

收起

文献

收起

参考文献引证文献

排序方式：

FU ZJ, WANG ZY, XU L, et al. HIF-1α-BNIP3-mediated mitophagy in tubular cells protects against renal ischemia/reperfusion injury. Redox Biol. 2020;36:101671.

JIANG Y, FAN X, YU Y, et al. USP13 Overexpression in BMSCs Enhances Anti-Apoptotic Ability and Guards Against Methylprednisolone-Induced Osteonecrosis in Rats. Stem Cells. 2024;43(1):sxae069.

LI J, LIN Q, SHAO X, et al. HIF1α-BNIP3-mediated mitophagy protects against renal fibrosis by decreasing ROS and inhibiting activation of the NLRP3 inflammasome. Cell Death Dis. 2023;14(3):200.

SHAO W, LI Z, WANG B, et al. Dimethyloxalylglycine Attenuates Steroid-Associated Endothelial Progenitor Cell Impairment and Osteonecrosis of the Femoral Head by Regulating the HIF-1α Signaling Pathway. Biomedicines. 2023;11(4):992.

ZHANG P, HA N, DAI Q, et al. Hypoxia suppresses osteogenesis of bone mesenchymal stem cells via the extracellular signal‑regulated 1/2 and p38‑mitogen activated protein kinase signaling pathways. Mol Med Rep. 2017;16(4):5515-5522.

CHEN W, WU P, YU F, et al. HIF-1α Regulates Bone Homeostasis and Angiogenesis, Participating in the Occurrence of Bone Metabolic Diseases. Cells. 2022;11(22):3552.

XU Y, JIANG Y, XIA C, et al. Stem cell therapy for osteonecrosis of femoral head: Opportunities and challenges. Regen Ther. 2020;15: 295-304.

HOUDEK MT, WYLES CC, PACKARD BD, et al. Decreased Osteogenic Activity of Mesenchymal Stem Cells in Patients With Corticosteroid-Induced Osteonecrosis of the Femoral Head. J Arthroplasty. 2016; 31(4):893-898.

YANG N, LI M, LI X, et al. MAGL blockade alleviates steroid-induced femoral head osteonecrosis by reprogramming BMSC fate in rat. Cell Mol Life Sci. 2024;81(1):418.

SAMAL JRK, RANGASAMI VK, SAMANTA S, et al. Discrepancies on the Role of Oxygen Gradient and Culture Condition on Mesenchymal Stem Cell Fate. Adv Healthc Mater. 2021;10(6):e2002058.

YAN C, SHI Y, YUAN L, et al. Mitochondrial quality control and its role in osteoporosis. Front Endocrinol (Lausanne). 2023;14:1077058.

NOWAK-STĘPNIOWSKA A, OSUCHOWSKA PN, FIEDOROWICZ H, et al. Insight in Hypoxia-Mimetic Agents as Potential Tools for Mesenchymal Stem Cell Priming in Regenerative Medicine. Stem Cells Int. 2022;2022: 8775591.

LU Z, JIANG L, LESANI P, et al. Nicotinamide Mononucleotide Alleviates Osteoblast Senescence Induction and Promotes Bone Healing in Osteoporotic Mice. J Gerontol A Biol Sci Med Sci. 2023; 78(2):186-194.

ABU-SHAHBA AG, GEBRAAD A, KAUR S, et al. Proangiogenic Hypoxia-Mimicking Agents Attenuate Osteogenic Potential of Adipose Stem/Stromal Cells. Tissue Eng Regen Med. 2020;17(4):477-493.

FENG X, XIANG Q, JIA J, et al. CircHGF suppressed cell proliferation and osteogenic differentiation of BMSCs in ONFH via inhibiting miR-25-3p binding to SMAD7. Mol Ther Nucleic Acids. 2022;28:99-113.

YANG Q, ZOU Y, WEI X, et al. PTP1B knockdown alleviates BMSCs senescence via activating AMPK-mediated mitophagy and promotes osteogenesis in senile osteoporosis. Biochim Biophys Acta Mol Basis Dis. 2023;1869(7):166795.

DONG Q, FEI X, ZHANG H, et al. Effect of Dimethyloxalylglycine on Stem Cells Osteogenic Differentiation and Bone Tissue Regeneration-A Systematic Review. Int J Mol Sci. 2024;25(7):3879.

ZHANG F, PENG W, WANG T, et al. Lnc Tmem235 promotes repair of early steroid-induced osteonecrosis of the femoral head by inhibiting hypoxia-induced apoptosis of BMSCs. Exp Mol Med. 2022;54(11): 1991-2006.

LU Y, LI Z, ZHANG S, et al. Cellular mitophagy: Mechanism, roles in diseases and small molecule pharmacological regulation. Theranostics. 2023;13(2):736-766.

WANG K, PENG X, ZHANG R, et al. COL6A3 enhances the osteogenic differentiation potential of BMSCs by promoting mitophagy in the osteoporotic microenvironment. Mol Biol Rep. 2024;51(1):206.

WU D, LIU L, FU S, et al. Osteostatin improves the Osteogenic differentiation of mesenchymal stem cells and enhances angiogenesis through HIF-1α under hypoxia conditions in vitro. Biochem Biophys Res Commun. 2022;606:100-107.

ZHAO F, VELDHUIS JJ, DUAN Y, et al. Low oxygen tension and synthetic nanogratings improve the uniformity and stemness of human mesenchymal stem cell layer. Mol Ther. 2010;18(5):1010-1018.

ZHAO D, LIU L, CHEN Q, et al. Hypoxia with Wharton’s jelly mesenchymal stem cell coculture maintains stemness of umbilical cord blood-derived CD34+ cells. Stem Cell Res Ther. 2018;9(1):158.

SONG S, ZHANG G, CHEN X, et al. HIF-1α increases the osteogenic capacity of ADSCs by coupling angiogenesis and osteogenesis via the HIF-1α/VEGF/AKT/mTOR signaling pathway. J Nanobiotechnology. 2023;21(1):257.

YI L, JU Y, HE Y, et al. Intraperitoneal injection of Desferal® alleviated the age-related bone loss and senescence of bone marrow stromal cells in rats. Stem Cell Res Ther. 2021;12(1):45.

LIN Q, LI S, JIANG N, et al. Inhibiting NLRP3 inflammasome attenuates apoptosis in contrast-induced acute kidney injury through the upregulation of HIF1A and BNIP3-mediated mitophagy. Autophagy. 2021;17(10):2975-2990.

BAI H, WANG Y, ZHAO Y, et al. HIF signaling: A new propellant in bone regeneration. Biomater Adv. 2022;138:212874.

HU Y, LOU B, WU X, et al. Comparative Study on In Vitro Culture of Mouse Bone Marrow Mesenchymal Stem Cells. Stem Cells Int. 2018; 2018:6704583.

WU J, NIU J, LI X, et al. Hypoxia induces autophagy of bone marrow-derived mesenchymal stem cells via activation of ERK1/2. Cell Physiol Biochem. 2014;33(5):1467-1474.

LIAO Z, JIN Y, CHU Y, et al. Single-cell transcriptome analysis reveals aberrant stromal cells and heterogeneous endothelial cells in alcohol-induced osteonecrosis of the femoral head. Commun Biol. 2022;5(1):324.

ZHAO D, ZHANG F, WANG B, et al. Guidelines for clinical diagnosis and treatment of osteonecrosis of the femoral head in adults (2019 version). J Orthop Translat. 2020;21:100-110.

ZHENG C, WU Y, XU J, et al. Exosomes from bone marrow mesenchymal stem cells ameliorate glucocorticoid-induced osteonecrosis of femoral head by transferring microRNA-210 into bone microvascular endothelial cells. J Orthop Surg Res. 2023;18(1):939.

MA S, LI S, ZHANG Y, et al. BMSC-Derived Exosomal CircHIPK3 Promotes Osteogenic Differentiation of MC3T3-E1 Cells via Mitophagy. Int J Mol Sci. 2023;24(3):2785.

MAS-BARGUES C, SANZ-ROS J, ROMÁN-DOMÍNGUEZ A, et al. Relevance of Oxygen Concentration in Stem Cell Culture for Regenerative Medicine. Int J Mol Sci. 2019;20(5):1195.

LIU F, YUAN Y, BAI L, et al. LRRc17 controls BMSC senescence via mitophagy and inhibits the therapeutic effect of BMSCs on ovariectomy-induced bone loss. Redox Biol. 2021;43:101963.

SUN K, JING X, GUO J, et al. Mitophagy in degenerative joint diseases. Autophagy. 2021;17(9):2082-2092.

LI G, JIAN Z, WANG H, et al. Irisin Promotes Osteogenesis by Modulating Oxidative Stress and Mitophagy through SIRT3 Signaling under Diabetic Conditions. Oxid Med Cell Longev. 2022;2022:3319056.

LI Y, FU G, GONG Y, et al. BMP-2 promotes osteogenic differentiation of mesenchymal stem cells by enhancing mitochondrial activity. J Musculoskelet Neuronal Interact. 2022;22(1):123-131.

XIANG K, REN M, LIU F, et al. Tobacco toxins trigger bone marrow mesenchymal stem cells aging by inhibiting mitophagy. Ecotoxicol Environ Saf. 2024;277:116392.

LEE JS, LEE JS, ROH HL, et al. Alterations in the differentiation ability of mesenchymal stem cells in patients with nontraumatic osteonecrosis of the femoral head: comparative analysis according to the risk factor. J Orthop Res. 2006;24(4):604-609.

2026年第30卷第1期

PDF下载

引用本文

BibTeX

文章信息

doi: 10.12307/2025.578

首发时间：2025-05-30
出版时间：2026-01-08

补充材料

相关文章

文章信息

作者

出版历史

录用日期：2025-03-08

基金

国家重点研发计划课题

解放军总医院青年自主创新科学基金项目

作者信息

^1.放军总医院第四医学中心骨科医学部研究所，北京市 100853

^2.放军医学院，北京市 100853

^3.放军总医院第四医学中心骨科医学部研究所，北京市 100853

通讯作者:

彭江: 翁土军，副研究员，解放军总医院第四医学中心骨科医学部研究所，北京市 100853

参考文献

分享链接

https://castjournals.cast.org.cn/joweb/zgzzgcyj/CN/10.12307/2025.578

分享至

全文二维码

扫描看全文

引用本文

BibTeX

本文的引用情况

2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

关闭全屏

BibTeX
EndNote
RefWorks
TxT