| 摘要: |
| 目的研究甘草中5种活性物质甘草酸(GA)、甘草苷(LQ)、异甘草苷(ILQ)、芹糖甘草苷(LA)和芹糖异甘草苷(ILA)对人脂肪间充质干细胞（human adipose-derived stem cells， hADSCs）抗衰老效果。方法用CCK-8法测定不同浓度甘草5种活性物质对hADSCs增殖的影响，选择5种化合物的适宜浓度对hADSCs进行培养，并用Ki67染色观察其增殖能力；利用细胞衰老β-半乳糖苷酶(senescence-associated β-galactosidase, SA-β-gal)染色法观察细胞衰老情况；用细胞内活性氧类(reactive oxygen species, ROS)试剂盒检测细胞内ROS含量；采用流式细胞仪检测hADSCs细胞周期；实时荧光定量聚合酶链式反应(real-time PCR)检测p16INK4a、p53、p21 mRNA的表达，测定端粒相对长度；对hADSCs进行成脂和成骨诱导培养，并分别利用油红O和茜素红染色观察脂滴和矿化结节。最后对化合物处理后细胞进行测序分析。结果CCK-8结果显示，甘草酸、甘草苷、异甘草苷、芹糖甘草苷以及芹糖异甘草苷在hADSCs实验中适宜浓度分别是3.125、3.125、3.125、3.125、6.25 μmol/L；在此浓度下，5种活性物质都能使Ki67阳性细胞数目显著增多，使β-半乳糖苷酶阳性数目显著减少；除芹糖甘草苷外能使细胞内ROS水平降低；芹糖异甘草苷能使G1期细胞比例下降；并且甘草酸、甘草苷和芹糖异甘草苷能使p16INK4a基因表达量下降，能减弱hADSCs成脂分化能力，促进细胞成骨分化能力。结论在甘草酸、甘草苷、异甘草苷、芹糖异甘草苷和芹糖甘草苷5种活性物质中，甘草酸的抗衰老效果最佳，其次是甘草苷和芹糖异甘草苷。 |
| 关键词: 甘草 甘草酸 人脂肪间充质干细胞 抗衰老 |
| DOI：10.12289/j.issn.1008-0392.20291 |
| 投稿时间：2020-07-04 |
| 基金项目:国家自然科学基金项目（31671539） |
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| Anti-aging effect of five active substancesfrom licorice on human adipose-derived stem cells |
| JIANG Rui-chan,LIU Hai-liang |
| (Ministry of Education Key Laboratory of Xinjiang Phytomedicine Resource and Utilization, College of Life Sciences,
Shihezi University, Shihezi 832003, Xinjiang Uygur Autonomous Region, China;Ministry of Education Key Laboratory of Xinjiang Phytomedicine Resource and Utilization, College of Life Sciences, Shihezi University, Shihezi 832003, Xinjiang Uygur Autonomous Region, China; Institute for Regenerative Medicine, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai 200123, China; Translational Medical Center for Stem Cell Therapy, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai 200123, China) |
| Abstract: |
| ObjectiveTo investigate the anti-aging effects of glycyrrhizic acid（GA), liquiritin（LQ), isoliquiritin（ILQ), liquiritin apioside（LA) and isoliquiritin apioside（ILA) from licorice on human adipose-derived stem cells（hADSCs). MethodsThe cell count kit（CCK-8) was used to detect the effect of different concentrations of five active substances on hADSCs, and the appropriate concentration was determined to treat hADSCs in cell culture. Cell proliferation ability was observed by Ki67 staining; cell senescence was observed by senescence-associated β-galactosidase（SA-β-gal) staining. Intracellular reactive oxygen species（ROS) kit was used to detect the content of reactive oxygen species. Cell cycle was observed by flow cytometry. Real-time fluorescence quantitative polymerase chain reaction（real-time RT-PCR) was used to detect the mRNA expression of p16INK4a, p53 and p21 and the relative length of telomeres. Adipogenic and osteoblastic induction cultures were performed on hADSCs, and lipid droplets and mineralized nodules were observed by oil red O and alizarin red staining, respectively. Finally, RNA sequencing was used to further study the changes of cellular pathways. ResultsCCK-8 results showed that the appropriate concentrations of GA, LQ, ILQ, LA and ILA in the hADSCs experiment were 3.125, 3.125, 3.125, 6.25 μmol/L, respectively. At this concentration, the positive number of Ki67 increased significantly, and the positive number of β-galactosidase decreased significantly. After treatment with these substances（except LA), the intracellular ROS levels were decreased. ILA treatment decreased the proportion of G1 phase cells. Treatment with GA, LQ and ILA reduced the expression of p16INK4a gene, reduced the adipogenic differentiation ability of hADSCs, and promoted the osteogenic differentiation ability of cells. ConclusionAmong the five active substances of licorice GA has the best anti-aging effect in hADSCs, followed by LQ and ILA. |
| Key words: licorice glycyrrhizic acid human adipose-derived stem cells anti-aging |
