
浏览全部资源
扫码关注微信
山东第二医科大学附属医院脊柱外科/潍坊市脊柱外科微创脊柱内镜重点实验室,山东 潍坊 261000
滕海军,E-mail:tenghaijun89@163.com
收稿:2025-02-11,
纸质出版:2026-02-25
移动端阅览
胡红涛,王胜,赵司顺等.脂肪间充质干细胞改善椎间盘髓核细胞衰老的机制研究[J].局解手术学杂志,2026,35(02):133-138.
HU Hong-tao,WANG Sheng,ZHAO Si-shun,et al.Mechanistic study of adipose-derived mesenchymal stem cell ameliorating nucleus pulposus cell senescence[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2026,35(02):133-138.
胡红涛,王胜,赵司顺等.脂肪间充质干细胞改善椎间盘髓核细胞衰老的机制研究[J].局解手术学杂志,2026,35(02):133-138. DOI: 10.11659/jjssx.02E025056.
HU Hong-tao,WANG Sheng,ZHAO Si-shun,et al.Mechanistic study of adipose-derived mesenchymal stem cell ameliorating nucleus pulposus cell senescence[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2026,35(02):133-138. DOI: 10.11659/jjssx.02E025056.
目的
2
探究脂肪间充质干细胞在氧化应激下衰老髓核细胞(NPC)中的作用及其介导凋亡调控的相关分子机制。
方法
2
根据处理条件不同将NPC分为空白组(NPC组)、实验组(NPC+ADSC组)、氧化损伤模型组(NPC+H
2
O
2
组)、实验+氧化损伤组(NPC+ADSC+H
2
O
2
组)。通过CCK-8法确定合适的H
2
O
2
浓度以构建氧化损伤模型,然后通过β-半乳糖苷酶染色分析各组NPC衰老程度。免疫荧光检测NPC组和NPC+ADSC组细胞中Ⅱ型胶原(Collagen Ⅱ)和聚集蛋白聚糖(Aggrecan)的表达;Western blot分析各组NPC中P53、P16、P-Rb、SIRT1、FoxO1、p-FoxO1、β-catenin、Bax、Bcl-2、Caspase-3的蛋白表达水平。
结果
2
与NPC+H
2
O
2
组相比,NPC+ADSC+H
2
O
2
组NPC衰老数量减少(
P
<
0.05)。NPC与ADSC共培养后细胞中Collagen Ⅱ、Aggrecan蛋白表达增加(
P
<
0.05)。Western blot检测发现,与ADSC共培养可以抑制H
2
O
2
诱导的NPC衰老相关蛋白(P53、P16及P-Rb)及线粒体凋亡相关蛋白(Bax及Caspase-3)的表达(
P
<
0.05),而SIRT1、p-FoxO1及Bcl-2蛋白表达升高(
P
<
0.05),FoxO1、β-catenin蛋白表达降低(
P
<
0.05)。
结论
2
ADSC可以修复氧化应激导致的NPC线粒体损伤,抑制其细胞衰老,该过程可能与调控SIRT1/FoxO1/β-catenin信号通路有关。
Objective
2
To explore the role of adipose-derived mesenchymal stem cells in nucleus pulposus cell (NPC) senescence induced by oxidative stress and the related molecular mechanisms mediating apoptosis regulation.
Methods
2
The NPCs were divided into the blank group (NPC group),experimental group (NPC+ADSC group),oxidative damage model group (NPC+H
2
O
2
group),experimental+oxidative damage group (NPC+ADSC+H
2
O
2
group) according to the different treatment conditions. The appropriate concentration of H
2
O
2
was determined by the CCK-8 method to construct the oxidative damage model, and then the senescence degree of NPCs in each group was analyzed by β-galactosidase staining. The expressions of Collagen Ⅱ and Aggrecan were detected in the NPC and NPC+ADSC groups by immunofluorescence. The expression levels of P53,P16,P-Rb,SIRT1,FoxO1,p-FoxO1,β-catenin,Bax,Bcl-2,Caspase-3 proteins in each group were detected by Western blot.
Results
2
Compared with the NPC+H
2
O
2
group, the number of aging NPCs in the NPC+ADSC+H
2
O
2
group was decreased (
P
<
0.05). After co-culture with ADSC,the expressions of Collagen Ⅱ and Aggrecan proteins in NPCs were increased(
P
<
0.05). Western blot results showed that co-culture with ADSC could inhibit the expression of aging-related proteins (P53, P16 and P-Rb) and mitochondrial apoptosis-related proteins (Bax and Caspase-3) in NPCs induced by H
2
O
2
(
P
<
0.05), and the expressions of SIRT1, p-FoxO1 and Bcl-2 proteins increased(
P
<
0.05), the expressions of FoxO1 and β-catenin proteins decreased(
P
<
0.05).
Conclusion
2
ADSC can repair oxidative stress-induced mitochondrial damage in NPC and inhibit its cellular senescence,the process that may be related to the regulation of the SIRT1/FoxO1/β-catenin signaling pathway.
Urits I , Burshtein A , Sharma M , et al . Low back pain,a comprehensive review: pathophysiology,diagnosis,and treatment [J]. Curr Pain Headache Rep , 2019 , 23 ( 3 ): 23 . doi: 10.1007/s11916-019-0757-1 http://dx.doi.org/10.1007/s11916-019-0757-1 .
Vergroesen PPA , Kingma I , Emanuel KS , et al . Mechanics and biology in intervertebral disc degeneration: a vicious circle [J]. Osteoarthritis Cartilage , 2015 , 23 ( 7 ): 1057 - 1070 . doi: 10.1016/j.joca.2015. 03.028 http://dx.doi.org/10.1016/j.joca.2015.03.028 .
Novais EJ , Tran VA , Miao J , et al . Comparison of inbred mouse strains shows diverse phenotypic outcomes of intervertebral disc aging [J]. Aging Cell , 2020 , 19 ( 5 ): e13148 . doi: 10.1111/acel.13148 http://dx.doi.org/10.1111/acel.13148 .
Nakamichi R , Asahara H . The transcription factors regulating inter-vertebral disc development [J]. JOR Spine , 2020 , 3 ( 1 ): e1081 . doi: 10.1002/jsp2.1081 http://dx.doi.org/10.1002/jsp2.1081 .
Eisenstein SM , Balain B , Roberts S . Current treatment options for intervertebral disc pathologies [J]. Cartilage , 2020 , 11 ( 2 ): 143 - 151 . doi: 10.1177/1947603520907665 http://dx.doi.org/10.1177/1947603520907665 .
Bowles RD , Setton LA . Biomaterials for intervertebral disc regeneration and repair [J]. Biomaterials , 2017 , 129 : 54 - 67 . doi: 10.1016/j.biomaterials.2017.03.013 http://dx.doi.org/10.1016/j.biomaterials.2017.03.013 .
Kang L , Liu S , Li J , et al . Parkin and Nrf2 prevent oxidative stress-induced apoptosis in intervertebral endplate chondrocytes via inducing mitophagy and anti-oxidant defenses [J]. Life Sci , 2020 , 243 : 117244 . doi: 10.1016/j.lfs.2019.117244 http://dx.doi.org/10.1016/j.lfs.2019.117244 .
Sakai D , Andersson GBJ . Stem cell therapy for intervertebral disc regeneration: obstacles and solutions [J]. Nat Rev Rheumatol , 2015 , 11 ( 4 ): 243 - 256 . doi: 10.1038/nrrheum.2015.13 http://dx.doi.org/10.1038/nrrheum.2015.13 .
Zhang Y , Wang Y , Zhou X , et al . Osmolarity controls the differentiation of adipose-derived stem cells into nucleus pulposus cells via histone demethylase KDM4B [J]. Mol Cell Biochem , 2020 , 472 ( 1-2 ): 157 - 171 . doi: 10.1007/s11010-020-03794-8 http://dx.doi.org/10.1007/s11010-020-03794-8 .
Wang B , Xu N , Cao L , et al . miR-31 from mesenchymal stem cell-derived extracellular vesicles alleviates intervertebral disc degeneration by inhibiting NFAT5 and upregulating the Wnt/β-catenin pathway [J]. Stem Cells Int , 2022 : 2164057 . doi: 10.1155/2022/2164057 http://dx.doi.org/10.1155/2022/2164057 .
Bhujel B , Shin HE , Choi DJ , et al . Mesenchymal stem cell-derived exosomes and intervertebral disc regeneration:review [J]. Int J Mol Sci , 2022 , 23 ( 13 ): 7306 . doi: 10.3390/ijms23137306 http://dx.doi.org/10.3390/ijms23137306 .
Zhao Y , Qin Y , Wu S , et al . Mesenchymal stem cells regulate inflammatory milieu within degenerative nucleus pulposus cells via p38 MAPK pathway [J]. Exp Ther Med , 2020 , 20 ( 5 ): 22 . doi: 10.3892/etm.2020.9150 http://dx.doi.org/10.3892/etm.2020.9150 .
Zhu L , Shi Y , Liu L , et al . Mesenchymal stem cells-derived exosomes ameliorate nucleus pulposus cells apoptosis via delivering miR-142-3p: therapeutic potential for intervertebral disc degenerative diseases [J]. Cell Cycle , 2020 , 19 ( 14 ): 1727 - 1739 . doi: 10.1080/15384101. 2020.1769301 http://dx.doi.org/10.1080/15384101.2020.1769301 .
Yoshikawa T , Ueda Y , Miyazaki K , et al . Disc regeneration therapy using marrow mesenchymal cell transplantation: a report of two case studies [J]. Spine , 2010 ,15, 35 ( 11 ): E475 - E480 . doi: 10.1097/BRS.0b013e3181cd2cf4 http://dx.doi.org/10.1097/BRS.0b013e3181cd2cf4 .
Li M , Li R , Yang S , et al . Exosomes derived from bone marrow mesenchymal stem cells prevent acidic pH-induced damage in human nucleus pulposus cells [J]. Med Sci Monit , 2020 , 26 : e922928 . doi: 10.12659/MSM.922928 http://dx.doi.org/10.12659/MSM.922928 .
Chen TS , Liao WY , Huang CW , et al . Adipose-derived stem cells preincubated with green tea EGCG enhance pancreatic tissue regeneration in rats with type 1 diabetes through ROS/Sirt1 signaling regulation [J]. Int J Mol Sci , 2022 , 23 ( 6 ): 3165 . doi: 10.3390/ijms23063165 http://dx.doi.org/10.3390/ijms23063165 .
Hsieh DJY , Marte L , Kuo WW , et al . Epigallocatechin-3-gallate preconditioned adipose-derived stem cells confer neuroprotection in aging rat brain [J]. Int J Med Sci , 2020 , 17 ( 13 ): 1916 - 1926 . doi: 10.7150/ijms.46696 http://dx.doi.org/10.7150/ijms.46696 .
Shi RF , Jin YP , Hu WW , et al . Exosomes derived from mmu_circ_0000250-modified adipose-derived mesenchymal stem cells promote wound healing in diabetic mice by inducing miR-128-3p/SIRT1-mediated autophagy [J]. Am J Physiol Cell Physiol , 2020 , 318 ( 5 ): C848 - C856 . doi: 10.1152/ajpcell.00041.2020 http://dx.doi.org/10.1152/ajpcell.00041.2020 .
Li M , Li RY , Yang SD , et al . Exosomes derived from bone marrow mesenchymal stem cells prevent acidic ph-induced damage in human nucleus pulposus cells [J]. Med Sci Monit , 2020 , 26 : e922928 . doi: 10.12659/MSM.922928 http://dx.doi.org/10.12659/MSM.922928 .
Zhuo Y , Wang H , Zou L , et al . SIRT1 attenuates apoptosis of nucleus pulposus cells by targeting interactions between LC3B and Fas under high-magnitude compression [J]. Oxid Med Cell Longev , 2021 : 2420969 . doi: 10.1155/2021/2420969 http://dx.doi.org/10.1155/2021/2420969 .
Santos BF , Grenho I , Martel PJ , et al . FOXO family isoforms [J]. Cell Death Dis , 2023 , 14 ( 10 ): 702 . doi: 10.1038/s41419-023-06177-1 http://dx.doi.org/10.1038/s41419-023-06177-1 .
Iyer S , Han L , Bartell SM , et al . Sirtuin1 (Sirt1) promotes cortical bone formation by preventing β-catenin sequestration by FoxO transcription factors in osteoblast progenitors [J]. J Biol Chem , 2014 , 289 ( 35 ): 24069 - 24078 . doi: 10.1074/jbc.M114.561803 http://dx.doi.org/10.1074/jbc.M114.561803 .
Yao H , Yao Z , Zhang S , et al . Upregulation of SIRT1 inhibits H 2 O 2 ‑induced osteoblast apoptosis via FoxO1/β‑catenin pathway [J]. Mol Med Rep , 2018 , 17 ( 5 ): 6681 - 6690 . doi: 10.3892/mmr. 2018.8657 http://dx.doi.org/10.3892/mmr.2018.8657 .
0
浏览量
2
下载量
0
CSCD
关联资源
相关文章
相关作者
相关机构
京公网安备11010802024621