摘要: |
目的探讨柚皮素在体外对人脂肪间充质干细胞(adipose-derived stem cell, ADSC)增殖的影响。方法采用CCK-8法检测柚皮素对ADSC细胞活性的影响,流式细胞术检测细胞周期,Western印迹法检测P16ink4a的表达水平,β-半乳糖苷酶染色试剂盒检测细胞衰老的情况,活性氧检测试剂盒检测细胞内活性氧类(ROS)含量,mRNA测序(RNA-seq)分析相关调控信号通路。结果CCK-8结果表明柚皮素在浓度为25μmol/L时能够明显促进ADSC的增殖(P<0.01),流式细胞术结果显示柚皮素能使ADSC快速进入S期,促进了细胞的增殖(P<0.05),Western印迹法检测结果显示柚皮素能够下调P16ink4a的表达(P<0.05),β-半乳糖苷酶染色检测结果显示柚皮素能够延缓细胞衰老(P<0.01),活性氧检测结果显示柚皮素能够清除细胞内活性氧的含量(P<0.01),RNA-seq结果表明柚皮素能够通过激活核苷酸结合寡聚结构域样受体(NOD-like receptors, NLRs)信号通路上调SOD2基因的表达促进hADSC的增殖。结论柚皮素能够促进ADSC的增殖,其相关机制可能与激活核苷酸结合寡聚结构域样受体信号通路上调SOD2基因的表达有关。 |
关键词: 柚皮素 脂肪间充质干细胞 细胞增殖 核苷酸结合寡聚结构域样受体信号通路 |
DOI:10.12289/j.issn.1008-0392.20367 |
投稿时间:2020-08-25 |
基金项目:国家重点研发计划项目(2020YFC2002800) |
|
Effect of naringenin on proliferation of adipose-derived stem cell |
WU Ya-ting,LIU Hai-liang |
(Ministry of Education Key Laboratory of Xinjiang Phytomedicine Resource and Utilization College of Life Sciences, Shihezi University, Shihezi 832003, Xinjiang Urgur Autonomous Region, China;Ministry of Education Key Laboratory of Xinjiang Phytomedicine Resource and Utilization College of Life Sciences, Shihezi University, Shihezi 832003, Xinjiang Urgur Autonomous Region, China; Institute for Regenerative Medicine, East Hospital, Tongji University School of Medicine, Shanghai 200123, China) |
Abstract: |
ObjectiveTo investigate the effect of naringenin (Nar) on proliferation of adipose-derived stem cell (ADSC). MethodsHuman ADSC was treated with Nar of different concentrations. Cell viability was measured by CCK-8 assay, cell cycle was detected by flowcytometry, the expression of P16ink4a was evaluated by Western blotting, aging cells were identified by β-galactosidase staining (SA-β-gal), reactive oxygen species (ROS) levels were examined by ROS reagent, the related regulate signaling pathway was determined by RNA-seq analysis. ResultsCCK-8 assay showed that 25μmol/L Nar increased the cell viability (P<0.01). Flow cytometry showed that Nar enhanced S phase and improved the proliferation of ADSC (P<0.05). Western blotting demonstrate that Nar decreased the expression of P16ink4a in ADSC (P<0.05). SA-β-gal result show that Nar inhibited the ADSC aging compared with control (P<0.01) and downregulate the ROS level (P<0.01). RNA-seq result showed that through NOD-like receptors (NLRs) signaling pathway Nar upregulate SOD2 mRNA expression and promoted the proliferation of ADSC. ConclusionNaringenin can promote the proliferation of adipose-derived stem cells, which is associated with upregulation of SOD2 mRNA expression through NOD-like receptors (NLRs) signaling pathway. |
Key words: naringenin adipose-derived stem cell cell proliferation NOD-like receptors signaling pathway |