SIRT1调控BMSCs成骨分化与H型血管生成促进骨质疏松骨缺损愈合的研究

doi:10.13701/j.cnki.kqyxyj.2024.09.006

口腔医学研究 ›› 2024, Vol. 40 ›› Issue (9): 785-792.DOI: 10.13701/j.cnki.kqyxyj.2024.09.006

SIRT1调控BMSCs成骨分化与H型血管生成促进骨质疏松骨缺损愈合的研究

杨启恒, 刘士博, 刘航航, 刘瑶, 邵京京, 罗恩^*

口腔疾病防治全国重点实验室国家口腔疾病临床医学研究中心四川大学华西口腔医院正颌及关节外科四川成都 610041

收稿日期:2024-04-12 出版日期:2024-09-28 发布日期:2024-09-25
通讯作者: ^*罗恩,E-mail:luoen521125@sina.com
作者简介:杨启恒(2000~),女,河北承德人,硕士在读,研究方向:口腔颌面外科。
基金资助:
国家自然科学基金面上项目(编号:82370932)四川省中央引导地方科技发展资金(编号:2023ZYD0107)四川省自然科学基金青年项目(编号:24NSFSC3073,2023NSFSC1393)

SIRT1 Regulates Osteogenic Differentiation of BMSCs and H-type Vessels Formation to Promote Healing of Osteoporotic Bone Defects

YANG Qiheng, LIU Shibo, LIU Hanghang, LIU Yao, SHAO Jingjing, LUO En^*

State Key Laboratory of Oral Diseases ＆ National Clinical Research Center for Oral Diseases, Department of Oral and Maxillofacial Surgery, West China Hospital of Stmatology, Sichuan University, Chengdu 610041, China

Received:2024-04-12 Online:2024-09-28 Published:2024-09-25

摘要/Abstract

摘要： 目的:探究沉默信息调节因子1(silent information regulator 1,SIRT1)通过调控骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)成骨分化与H型血管生成对老年骨质疏松状态下骨缺损愈合的作用,并探讨其具体分子机制。方法:利用免疫组织化学染色检测小鼠骨组织中SIRT1表达的增龄性改变。使用SIRT1激活剂SRT1720与抑制剂EX527作用于BMSCs, 实时荧光定量逆转录聚合酶链反应(quantitative real-time PCR,RT-qPCR)与Western blot检测成骨标志物以及促H型血管形成因子的表达变化,免疫荧光染色与Western blot检测上述过程中Wnt/连环蛋白β(β-catenin)信号通路变化。构建老年骨质疏松骨缺损模型,SRT1720与EX527作用的同时,分别给予Wnt信号通路抑制剂XAV939与激活剂CHIR-99021,Micro-CT、苏木精-伊红(hematoxylin-eosin staining,HE)/Masson染色与免疫荧光染色检测各组骨缺损区域新骨形成与H型血管含量差异。结果:小鼠骨组织中SIRT1表达呈现增龄性下调趋势。SIRT1能够促进成骨因子碱性磷酸酶(alkaline phosphatase, ALP)、矮小相关转录因子2(Runt-related transcription factor 2, RUNX2)和促H型血管形成因子slit同源物3(slit homolog 3, SLIT3)、血管内皮生长因子(vascular endothelial growth factor, VEGF)的表达,并促进老年骨质疏松骨缺损区域内H型血管形成与骨再生,且上述作用由Wnt/β-catenin信号通路介导。结论:SIRT1能够通过调控Wnt/β-catenin信号通路促进BMSCs成骨分化与H型血管生成,进而促进老年骨质疏松小鼠的骨缺损愈合。

关键词: 沉默信息调节因子1, Wnt/β-catenin信号通路, H型血管, 老年骨质疏松, 骨缺损

Abstract: Objective: To investigate the role of silent information regulator 1 (SIRT1) on H-type vessels regeneration and bone defect healing in senile osteoporotic state by regulating osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and to explore its specific molecular mechanism. Methods: Immunohistochemical staining was used to detect age-related changes in SIRT1 expression in mouse bone tissue. BMSCs were treated with SIRT1 activator SRT1720 and inhibitor EX527, and the changes in the expression of osteogenic markers and H-type angiogenic factors were detected by RT-qPCR and Western Blot, and the changes in the Wnt/β-catenin signaling pathway in the above process were detected by immunofluorescence staining and Western Blot. A model of osteoporotic bone defects was constructed in the elderly, and SRT1720 and EX527 were administered with XAV939, an inhibitor of Wnt signaling pathway, and CHIR-99021, an activator of Wnt signaling pathway, respectively. The differences in the formation of new bone and H-vessel content of the defects were detected by Micro-CT, HE/Masson staining, and immunofluorescent staining in the respective groups. Results: The expression of SIRT1 showed an age-increasing down-regulation trend in mouse bone tissues. SIRT1 was able to promote the expression of alkaline phosphatase (ALP), Runt-related transcription factor 2 (RUNX2), and the pro-H-type angiogenic factors slit homolog 3 (SLIT3) and vascular endothelial growth factor (VEGF), and able to promote the H-type angiogenesis and bone regeneration in the region of osteoporotic bone defects in old age, which was mediated by the Wnt/β-catenin signaling pathway. Conclusion: SIRT1 can promote the osteogenic differentiation and H-type angiogenesis of BMSCs by regulating the Wnt/β-catenin signaling pathway, which in turn promotes the healing of bone defects in the osteoporotic state of aged mice.

Key words: SIRT1, Wnt/β-catenin signaling pathway, H-type vessels, senile osteoporosis, bone defect

杨启恒, 刘士博, 刘航航, 刘瑶, 邵京京, 罗恩. SIRT1调控BMSCs成骨分化与H型血管生成促进骨质疏松骨缺损愈合的研究[J]. 口腔医学研究, 2024, 40(9): 785-792.

YANG Qiheng, LIU Shibo, LIU Hanghang, LIU Yao, SHAO Jingjing, LUO En. SIRT1 Regulates Osteogenic Differentiation of BMSCs and H-type Vessels Formation to Promote Healing of Osteoporotic Bone Defects[J]. Journal of Oral Science Research, 2024, 40(9): 785-792.

参考文献

[1] 贺丽英,孙蕴,要文娟,等. 2010-2016年中国老年人骨质疏松症患病率Meta分析[J].中国骨质疏松杂志,2016,22(12): 1590-1596.
[2] Filipowska J, Tomaszewski KA, Niedzwiedzki L, et al. The role of vasculature in bone development, regeneration and proper systemic functioning [J]. Angiogenesis, 2017, 20(3): 291-302.
[3] Fan D, Lu J, Yu N, et al. Curcumin prevents diabetic osteoporosis through promoting osteogenesis and angiogenesis coupling via NF-kappaB signaling [J]. Evid Based Complement Alternat Med, 2022, 2022: 4974343.
[4] Jing H, Liao L, Su X, et al. Declining histone acetyltransferase GCN5 represses BMSC-mediated angiogenesis during osteoporosis [J]. FASEB J, 2017, 31(10): 4422-4433.
[5] Kusumbe AP, Ramasamy SK, Adams RH. Coupling of angiogenesis and osteogenesis by a specific vessel subtype in bone [J]. Nature, 2014, 507(7492): 323-328.
[6] Avilkina V, Chauveau C, Ghali MO. Sirtuin function and metabolism: Role in pancreas, liver, and adipose tissue and their crosstalk impacting bone homeostasis [J]. Bone, 2022, 154: 116232.
[7] Chen W, Chen X, Chen AC, et al. Melatonin restores the osteoporosis-impaired osteogenic potential of bone marrow mesenchymal stem cells by preserving SIRT1-mediated intracellular antioxidant properties [J]. Free Radic Biol Med, 2020, 146: 92-106.
[8] Lv N, Zhou Z, Hou M, et al. Research progress of vascularization strategies of tissue-engineered bone [J]. Front Bioeng Biotechnol, 2023, 11: 1291969.
[9] Stahl A, Yang YP. Regenerative approaches for the treatment of large bone defects [J]. Tissue Eng Part B Rev, 2021, 27(6): 539-547.
[10] Chen J, Li M, Liu AQ, et al. Gli1⁺ cells couple with type H vessels and are required for type H vessel formation [J]. Stem Cell Reports, 2020, 15(1): 110-124.
[11] Xie H, Cui Z, Wang L, et al. PDGF-BB secreted by preosteoclasts induces angiogenesis during coupling with osteogenesis [J]. Nat Med, 2014, 20(11): 1270-1278.
[12] Mercken EM, Mitchell SJ, Martin-Montalvo A, et al. SRT2104 extends survival of male mice on a standard diet and preserves bone and muscle mass [J]. Aging Cell, 2014, 13(5): 787-796.
[13] Song J, Li J, Yang F, et al. Nicotinamide mononucleotide promotes osteogenesis and reduces adipogenesis by regulating mesenchymal stromal cells via the SIRT1 pathway in aged bone marrow [J]. Cell Death Dis, 2019, 10(5): 336.
[14] Lemieux ME, Yang X, Jardine K, et al. The Sirt1 deacetylase modulates the insulin-like growth factor signaling pathway in mammals [J]. Mech Ageing Dev, 2005, 126(10): 1097-1105.
[15] Maeda K, Kobayashi Y, Koide M, et al. The regulation of bone metabolism and disorders by Wnt signaling [J]. Int J Mol Sci, 2019, 20(22):5525.
[16] Nusse R, Clevers H. Wnt/beta-catenin signaling, disease, and emerging therapeutic modalities [J]. Cell, 2017, 169(6): 985-999.
[17] Sun Q, Liu S, Feng J, et al. Current status of microRNAs that target the Wnt signaling pathway in regulation of osteogenesis and bone metabolism: A review [J]. Med Sci Monit, 2021, 27: e929510.
[18] Wong SK, Mohamad NV, Jayusman PA, et al. A review on the crosstalk between insulin and Wnt/beta-catenin signalling for bone health [J]. Int J Mol Sci, 2023, 24(15):12441.

SIRT1调控BMSCs成骨分化与H型血管生成促进骨质疏松骨缺损愈合的研究

SIRT1 Regulates Osteogenic Differentiation of BMSCs and H-type Vessels Formation to Promote Healing of Osteoporotic Bone Defects

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

[1]	刘苗, 李风兰. 数字化技术在上颌骨缺损赝复体修复中的应用[J]. 口腔医学研究, 2024, 40(5): 385-388.
[2]	刘纯, 王静文, 赵蕤, 赵欣欣, 赵熠. 节段性下颌骨切除腓骨重建后全口义齿修复1例[J]. 口腔医学研究, 2023, 39(4): 375-377.
[3]	张舟, 梁想, 崔浩, 朱羿霏, 屠军波, 那思家. 人骨髓间充质干细胞/细胞外基质复合体用于骨缺损再生修复的研究[J]. 口腔医学研究, 2022, 38(4): 372-378.
[4]	卢嘉蕊, 权晶晶. 骨缺损动物模型的研究进展[J]. 口腔医学研究, 2021, 37(9): 783-786.
[5]	管其帅, 许贺, 于甜甜, 许胜, 周文娟, 柳忠豪. 缺牙间隙对美学区水平骨增量延期种植的临床效果影响研究[J]. 口腔医学研究, 2021, 37(7): 607-611.
[6]	王鹏程, 陈春晖, 童熹, 傅新海. β-TCP植入与口腔修复膜覆盖治疗对颌骨囊肿术后骨缺损的修复效果分析[J]. 口腔医学研究, 2021, 37(3): 260-263.
[7]	司超, 邹馨颖, 刘悦, 张佩佩, 徐依山, 郝艺帆, 公柏娟. 载碱性成纤维细胞生长因子温敏水凝胶对大鼠骨髓间充质干细胞体外成骨分化的作用研究[J]. 口腔医学研究, 2021, 37(12): 1139-1144.
[8]	肖健平, 李丽丽, 李厚轩, 谭葆春. 自体新鲜脱矿牙齿移植物在不同部位成骨效果的动物实验[J]. 口腔医学研究, 2021, 37(12): 1145-1149.
[9]	谢惠兰, 方芳, 林毅. Chir99021调控Wnt/β-catenin信号通路干预干细胞成骨/成牙分化的研究[J]. 口腔医学研究, 2021, 37(12): 1150-1155.
[10]	许来俊, 李继遥. 结冷胶复合材料在骨缺损再生领域的研究进展[J]. 口腔医学研究, 2020, 36(6): 512-515.
[11]	左志刚, 李洪发, 王悦, 郑朝, 杨子靓, 刘珊, 刘大勇. SIRT1对正畸静压力刺激下牙周膜干细胞成骨分化的调控作用及机制[J]. 口腔医学研究, 2020, 36(11): 1069-1073.
[12]	薛国平, 刘青梅. 低水平激光治疗应用于颌骨骨再生的研究进展[J]. 口腔医学研究, 2019, 35(9): 830-832.
[13]	李智聪, 邹瑞, Hye-Jin Tak, Sang-Hwy Lee, 朴正国. 抑制Wnt/β-catenin信号通路对上唇发育的影响[J]. 口腔医学研究, 2019, 35(7): 695-700.
[14]	李冀寅, 贺平. 自体牙骨移植材料在牙槽骨缺损修复中的应用进展[J]. 口腔医学研究, 2019, 35(3): 212-214.
[15]	郑钧元, 何俐, 胡图强, 王美钧. 铒激光治疗老年患者前磨牙牙周角型骨缺损的半年疗效观察1例[J]. 口腔医学研究, 2019, 35(3): 307-308.