熊果酸通过缓解骨髓间充质干细胞衰老改善骨质疏松的研究

doi:10.13591/j.cnki.kqyx.2026.05.001

摘要/Abstract

摘要：

目的探讨熊果酸（ursolic acid，UA）能否通过缓解骨髓间充质干细胞（bone marrow mesenchymal stem cells，BMMSCs）衰老改善骨质疏松相关颌骨骨损伤。方法建立去卵巢（ovariectomy，OVX）骨质疏松小鼠模型，UA干预后采用Micro-CT评估股骨与颌骨骨小梁微结构；结合多数据库整合分析、KEGG通路富集、CellAge数据库及机器学习方法筛选UA作用于骨质疏松的关键衰老相关基因，基于骨质疏松单细胞转录组数据，分析关键基因在不同细胞类型中的表达特征，并采用SenMayo衰老相关基因集对不同细胞类型进行衰老评分；体外采用H₂O₂诱导BMMSCs衰老并给予UA处理，通过SA-β-gal、ALP/ARS染色及RT-PCR评价衰老表型与成骨能力。结果 Micro-CT结果显示，UA可显著改善骨质疏松模型小鼠股骨及颌骨的骨小梁微结构。生物信息学分析表明，UA作用于骨质疏松的潜在靶点显著富集于p53信号通路及细胞衰老相关通路。单细胞转录组分析结果显示，多种关键衰老相关基因在BMMSCs中表达，且基于SenMayo衰老相关基因集的多种评分方法一致提示在骨质疏松样本中BMMSCs具有较高的衰老水平。体外实验结果表明，UA可显著缓解H₂O₂诱导的BMMSCs成骨抑制及衰老表型。结论 UA可通过缓解 BMMSCs衰老改善骨质疏松相关骨损伤，对颌骨具有潜在骨保护作用。

关键词: 熊果酸, 骨质疏松, 颌骨, 骨髓间充质干细胞, 细胞衰老

Abstract:

Objective To investigate whether ursolic acid （UA） can ameliorate osteoporosis-related jaw bone injuries by alleviating the senescence of bone marrow mesenchymal stem cells （BMMSCs）. Methods An ovariectomized osteoporotic mouse model was established. After UA intervention，Micro-CT was used to evaluate the trabecular microstructure of the femur and jaw bone. Integrated multidatabase analysis，KEGG pathway enrichment，the CellAge database，and machine learning methods were employed to screen key senescence-related genes through which UA exerts its effects on osteoporosis. Based on single cell transcriptomic data from osteoporotic bone，the expression profiles of these key genes across different cell types were analyzed，and senescence scores for distinct cell populations were calculated using the SenMayo senescence-associated gene set. In vitro，BMMSCs were induced to senesce by H₂O₂ and then treated with UA. Senescence phenotypes and osteogenic capacity were assessed through SA-β-gal staining，ALP/ARS staining，and RT-PCR. Results Micro-CT analysis demonstrated that UA significantly improved trabecular bone micro-architecture in both the femur and jaw of osteoporotic mice. Bioinformatics analyses revealed that the potential targets of UA in osteoporosis were significantly enriched in the p53 signaling pathway and cellular senescence-related pathways. Single-cell transcriptomic analysis showed that multiple key senescence-related genes were expressed in BMMSCs，and senescence scoring based on the SenMayo gene set consistently indicated a higher level of senescence in BMMSCs from osteoporotic samples. Experiments further demonstrated that UA significantly alleviated hydrogen peroxide-induced senescent phenotypes in BMMSCs. Conclusion UA can ameliorate osteoporosis-associated bone injuries by alleviating senescence in BMMSCs，thereby exhibiting a potential bone-protective effect on the jaw.

Key words: ursolic acid, osteoporosis, jaw bone, bone marrow mesenchymal stem cells, cellular senescence

中图分类号:

R782.4

孙雪雨, 赵炜业, 张柯佳, 曹丹, 张瀚文, 严斌. 熊果酸通过缓解骨髓间充质干细胞衰老改善骨质疏松的研究[J]. 口腔医学, 2026, 46(5): 321-329.

SUN Xueyu, ZHAO Weiye, ZHANG Kejia, CAO Dan, ZHANG Hanwen, YAN Bin. Study on ursolic acid ameliorating osteoporosis by alleviating senescence of bone marrow mesenchymal stem cells[J]. Stomatology, 2026, 46(5): 321-329.

图/表 9

表1

图1

图2

图3

图4

图5

图6

图7

图8

参考文献 36

[1]	Antoniou T, MacDonald EM, Yao Z, et al. A population-based study of the risk of osteoporosis and fracture with dutasteride and finasteride[J]. BMC Musculoskelet Disord, 2018, 19(1): 160.
[2]	Rachner TD, Khosla S, Hofbauer LC. Osteoporosis: Now and the future[J]. Lancet, 2011, 377(9773): 1276-1287.
[3]	Shu XY, Mei M, Ma LQ, et al. Postmenopausal osteoporosis is associated with elevated aldosterone/renin ratio[J]. J Hum Hypertens, 2018, 32(7): 524-530.
[4]	Oliveira ML, Pedrosa EFNC, Cruz AD, et al. Relationship between bone mineral density and trabecular bone pattern in postmenopausal osteoporotic Brazilian women[J]. Clin Oral Investig, 2013, 17(8): 1847-1853.
[5]	Lemos CAA, de Oliveira AS, Faé DS, et al. Do dental implants placed in patients with osteoporosis have higher risks of failure and marginal bone loss compared to those in healthy patients?A systematic review with meta-analysis[J]. Clin Oral Investig, 2023, 27(6): 2483-2493.
[6]	Zaidi M. Skeletal remodeling in health and disease[J]. Nat Med, 2007, 13(7): 791-801.
[7]	Bianco P, Sacchetti B, Riminucci M. Stem cells in skeletal physiology and endocrine diseases of bone[J]. Endocr Dev, 2011, 21: 91-101.
[8]	Liu GH, Xiao GZ, Su JC, et al. Editorial: Tissue stem cells during trauma: From basic biology to translational medicine[J]. Front Cell Dev Biol, 2022, 10: 914582.
[9]	Teven CM, Rossi MT, Shenaq DS, et al. Bone morphogenetic protein-9 effectively induces osteogenic differentiation of reversibly immortalized calvarial mesenchymal progenitor cells[J]. Genes Dis, 2015, 2(3): 268-275.
[10]	Li Y, Fan LK, Hu J, et al. miR-26a rescues bone regeneration deficiency of mesenchymal stem cells derived from osteoporotic mice[J]. Mol Ther, 2015, 23(8): 1349-1357.
[11]	Hu ZB, Zhang LJ, Wang H, et al. Targeted silencing of miRNA-132-3p expression rescues disuse osteopenia by promoting mesenchymal stem cell osteogenic differentiation and osteogenesis in mice[J]. Stem Cell Res Ther, 2020, 11(1): 58.
[12]	Zupan J, Strazar K, Kocijan R, et al. Age-related alterations and senescence of mesenchymal stromal cells: Implications for regenerative treatments of bones and joints[J]. Mech Ageing Dev, 2021, 198: 111539.
[13]	Weng ZJ, Wang YG, Ouchi T, et al. Mesenchymal stem/stromal cell senescence: Hallmarks, mechanisms, and combating strategies[J]. Stem Cells Transl Med, 2022, 11(4): 356-371.
[14]	Neri S, Borzì RM. Molecular mechanisms contributing to mesenchymal stromal cell aging[J]. Biomolecules, 2020, 10(2): 340.
[15]	Wang BS, Han J, Elisseeff JH, et al. The senescence-associated secretory phenotype and its physiological and pathological implications[J]. Nat Rev Mol Cell Biol, 2024, 25(12): 958-978.
[16]	Pan Y, Gu ZZ, Lyu YS, et al. Link between senescence and cell fate: Senescence-associated secretory phenotype and its effects on stem cell fate transition[J]. Rejuvenation Res, 2022, 25(4): 160-172.
[17]	Farr JN, Roforth MM, Fujita K, et al. Effects of age and estrogen on skeletal gene expression in humans as assessed by RNA sequencing[J]. PLoS One, 2015, 10(9): e0138347.
[18]	Wang Z, Li XH, Yang JX, et al. Single-cell RNA sequencing deconvolutes the heterogeneity of human bone marrow-derived mesenchymal stem cells[J]. Int J Biol Sci, 2021, 17(15): 4192-4206.
[19]	Qiu X, Liu Y, Shen H, et al. Single-cell RNA sequencing of human femoral head in vivo[J]. Aging, 2021, 13(11): 15595-15619.
[20]	Saul D, Kosinsky RL, Atkinson EJ, et al. A new gene set identifies senescent cells and predicts senescence-associated pathways across tissues[J]. Nat Commun, 2022, 13(1): 4827.
[21]	Wen JH. Ursolic acid: Pharmacokinetics process in vitro and in vivo, a mini review[J]. Arch Pharm, 2019, 352(3): e1800222.
[22]	赵玉芹, 张成杰, 相淑芳. 熊果酸衍生物对糖尿病骨质疏松骨重塑的影响[J]. 滨州医学院学报, 2019, 42(4): 291-294.
[23]	Poudel K, Gautam M, Maharjan S, et al. Dual stimuli-responsive ursolic acid-embedded nanophytoliposome for targeted antitumor therapy[J]. Int J Pharm, 2020, 582: 119330.
[24]	Qian ZZ, Wang XH, Song Z, et al. A phase I trial to evaluate the multiple-dose safety and antitumor activity of ursolic acid liposomes in subjects with advanced solid tumors[J]. Biomed Res Int, 2015, 2015: 809714.
[25]	Prasad S, Yadav VR, Sung B, et al. Ursolic acid inhibits growth and metastasis of human colorectal cancer in an orthotopic nude mouse model by targeting multiple cell signaling pathways: Chemosensitization with capecitabine[J]. Clin Cancer Res, 2012, 18(18): 4942-4953.
[26]	Ahmad Dar B, Lone AM, Shah WA, et al. Synthesis and screening of ursolic acid-benzylidine derivatives as potential anti-cancer agents[J]. Eur J Med Chem, 2016, 111: 26-32.
[27]	Checker R, Sandur SK, Sharma D, et al. Potent anti-inflammatory activity of ursolic acid, a triterpenoid antioxidant, is mediated through suppression of NF-κB, AP-1 and NF-AT[J]. PLoS One, 2012, 7(2): e31318.
[28]	郑茜, 刘萍萍, 顾煜婕, 等. 熊果酸干预人牙周膜干细胞成骨分化的效应[J]. 中国组织工程研究, 2025, 29(1): 80-86.
[29]	Hao LY, Li J, Tian YW, et al. Changes in the microRNA profile of the mandible of ovariectomized mice[J]. Cell Physiol Biochem, 2016, 38(4): 1267-1287.
[30]	Luan MZ, Wang HY, Wang JZ, et al. Advances in anti-inflammatory activity, mechanism and therapeutic application of ursolic acid[J]. Mini Rev Med Chem, 2022, 22(3): 422-436.
[31]	Chen L, Li F, Ni JH, et al. Ursolic acid alleviates lupus nephritis by suppressing SUMO1-mediated stabilization of NLRP3[J]. Phytomedicine, 2024, 130: 155556.
[32]	Liu SJ, Guo BD, Gao QH, et al. Ursolic acid alleviates chronic prostatitis regulating NLRP3 inflammasome-mediated Caspase-1/GSDMD pyroptosis pathway[J]. Phytother Res, 2024, 38(1): 82-97.
[33]	Lei P, Li ZY, Hua QW, et al. Ursolic acid alleviates neuroinflammation after intracerebral hemorrhage by mediating microglial pyroptosis the NF-κB/NLRP3/GSDMD pathway[J]. Int J Mol Sci, 2023, 24(19): 14771.
[34]	Li HK, Xu N, Li SC, et al. Exosomes derived from umbilical cord mesenchymal stem cells alleviate jaw bone marrow mesenchymal stem cells senescence and restore osteogenic differentiation potential[J]. Stem Cell Res Ther, 2025, 16(1): 475.
[35]	Li PS, Ou QM, Shi ST, et al. Immunomodulatory properties of mesenchymal stem cells/dental stem cells and their therapeutic applications[J]. Cell Mol Immunol, 2023, 20(6): 558-569.
[36]	Jin H, Zhang YY, Hou CY, et al. Artesunate reverses BMSC senescence and ameliorates osteoporosis by upregulating NSD2[J]. Eur J Pharmacol, 2026, 1011: 178447.

引物	Forward primer 5'—3'	Reverse primer 5'—3'
Alp	ATAACGAGATGCCACCAGAGG	TTCCACATCAGTTCTGTTCTTCG
Runx2	AGAATGGACGTGCCCCCTA	CTGGGGAAGCAGCAACACTA
Osx	AAAGGAGGCACAAAGAAGC	CAGGAAATGAGTGAGGGAAG
Ocn	TTCTGCTCACTCTGCTGACCC	CTGATAGCTCGTCACAAGCAGG
Gapdh	TCCATGACAACTTTGGTATCG	TGTAGCCAAATTCGTTGTCA