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段嘉豪,李兆勇,陈龙,刘恩旭,李硕夫,杨雷,杨少锋.脂肪间充质干细胞修复氧化损伤髓核细胞机制研究[J].湖南中医药大学学报英文版,2023,43(8):1408-1414.[Click to copy
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| 脂肪间充质干细胞修复氧化损伤髓核细胞机制研究 |
| 段嘉豪,李兆勇,陈龙,刘恩旭,李硕夫,杨雷,杨少锋 |
| (湖南中医药大学第一附属医院, 湖南 长沙 410007) |
| 摘要: |
| 目的 探究脂肪间充质干细胞（adipose-derived mesenchymal stem cell，ADSC）修复氧化损伤髓核细胞（nucleus pulposus cell，NPC）的作用机制。方法 实验分为NPC组、ADSC组、NPC+ADSC组、H2O2+NPC+ADSC组、H2O2+NPC组5组。采用H2O2构建NPC氧化损伤模型；SA-β-Gal染色分析NPC衰老程度；流式细胞仪检测NPC氧化损伤水平；Western blot检测ADSC中Collagen Ⅱ、Aggrecan蛋白表达水平，以及NPC中Collagen Ⅱ、p53、p21、TGF-β1、Smad2、p-Smad2蛋白表达水平；qPCR检测ADSC中Collagen Ⅱ、Aggrecan mRNA表达水平，以及NPC中Collagen Ⅱ、p53、p21、TGF-β1、Smad2 mRNA表达。结果 与H2O2+NPC组相比较，H2O2+NPC+ADSC组的NPC衰老数量减少（P<0.05），NPC氧化损伤减轻（P<0.05），p53、p21蛋白和基因表达下降（P<0.05），Collagen Ⅱ、TGF-β1、p-Smad2蛋白和基因表达升高（P<0.05）。与ADSC组相比，NPC+ADSC组Collagen Ⅱ、Aggrecan基因和蛋白表达升高（P<0.05）。结论 ADSC与NPC共培养不仅可以促进ADSC类髓核化，而且ADSC可修复氧化损伤的NPC，抑制其衰老，该过程可能与调控TGF-β1/Smad2信号通路有关。 |
| 关键词: 脂肪间充质干细胞 髓核细胞 氧化损伤 细胞衰老 TGF-β1/Smad2信号通路 |
| DOI：10.3969/j.issn.1674-070X.2023.08.010 |
| Received:March 27, 2023 |
| 基金项目:国家自然科学基金项目（82174402）；湖南省自然科学基金项目（2023JJ60345；2023JJ60342）；湖南省教育厅科学研究项目（21C0233）；湖南省卫健委科研计划项目（202204074858）；湖南省研究生科研创新课题项目（CX20220807）；湖南中医药大学校级项目（2021XJJJ041）。 |
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| Mechanism of adipose-derived mesenchymal stem cells repairing oxidatively damaged nucleus pulposus cells |
| DUAN Jiahao,LI Zhaoyong,CHEN Long,LIU Enxu,LI Shuofu,YANG Lei,YANG Shaofeng |
| (The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China) |
| Abstract: |
| Objective To investigate the mechanism of adipose-derived mesenchymal stem cell (ADSC) repairing oxidatively damaged nucleus pulposus cell (NPC). Methods Five groups were divided as NPC group, ADSC group, NPC+ADSC group, H2O2+NPC+ADSC group, and H2O2+NPC group. Hydrogen peroxide (H2O2) was used to construct the oxidative damage model of NPC; SA-β-Gal staining was performed to analyze the degree of senescence in NPC; flow cytometer was used to determine the level of oxidative damage in NPC; Western blot was used to check the protein expressions of Collagen Ⅱ and Aggrecan in ADSC, and the protein expressions of Collagen Ⅱ, p53, p21, TGF-β1, Smad2, and p-Smad2 in NPC; qPCR was carried out to determine the mRNA expressions of Collagen Ⅱ and Aggrecan in ADSC, and the mRNA expressions of Collagen Ⅱ, p53, p21, TGF-β1, and Smad2 in NPC. Results Compared with H2O2+NPC group, H2O2+NPC+ADSC group showed a reduced number of senescent NPC (P<0.05), attenuated oxidative damage to NPC (P<0.05), decreased protein and mRNA expressions of p53 and p21 (P<0.05), and elevated protein and mRNA expressions of Collagen II, TGF-β1, and p-Smad2 (P<0.05). Compared with ADSC group, the protein and mRNA expressions of Collagen Ⅱ and Aggrecan were elevated in NPC+ADSC group (P<0.05). Conclusion Co-culture of ADSC and NPC can promote ADSC-like myelination, and ADSC can repair oxidatively damaged NPC and inhibit their senescence, which may be related to the regulation of TGF-β1/Smad2 pathway. |
| Key words: adipose-derived mesenchymal stem cells nucleus pulposus cells oxidative damage cell senescence TGF-β1/Smad2 |
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