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| 蒙花苷清除衰老的骨髓间充质干细胞发挥抗衰老作用* |
| Linarin play an anti-aging role by directed elimination bone marrow mesenchymal stem cells |
| 投稿时间：2020-02-23 修订日期：2020-03-02 |
| DOI： |
| 中文关键词: 蒙花苷 抗衰老 骨髓间充质干细胞 细胞衰老 |
| 英文关键词: linarin anti-aging bone marrow mesenchymal stem cells cell senescence |
| 基金项目:国家自然科学基金项目（面上项目，重点项目，重大项目） |
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| 中文摘要: |
| 目的:探讨蒙花苷(Linarin)是否可以清除衰老的大鼠骨髓间充质干细胞发挥抗衰老作用.方法: 取4周龄80 g-100 g的雄性SD大鼠大腿骨髓腔内的骨髓间充质干细胞,用含10 %胎牛血清的DMEM/F12培养基培养至三代,使用D-gal诱导及多次复制传代使骨髓间充质干细胞衰老并用SA-β-gal染色来确认细胞处于衰老状态, 通过蛋白印迹检测对照组与衰老组中细胞保护性蛋白sirt1、sirt6及衰老相关蛋白p16、p21、p53的表达水平.之后,通过cck-8检测蒙花苷的细胞毒性,找到目的浓度,用不同浓度处理衰老骨髓间充质干细胞.最后,通过蛋白印迹分析、免疫荧光检测对照组与处理组衰老相关蛋白表达情况.观察蒙花苷能否可以清除衰老的大鼠骨髓间充质干细胞发挥抗衰老作用.结果:与正常细胞相比,衰老的骨髓间充质干细胞细胞保护性蛋白sirt1、sirt6降低及衰老相关蛋白p16、p21、p53明显增高, 衰老细胞中与DNA损伤相关的细胞核内蛋白γ-H2AX表达明显增加.而蒙花苷处理后,衰老细胞组衰老相关蛋白p16、p21、p53明显降低,γ-H2AX蛋白阳性细胞明显减少.结论: 蒙花苷可以剂量依赖性的清除衰老的大鼠骨髓间充质干细胞发挥抗衰老作用. |
| 英文摘要: |
| Objective: To investigate whether Linarin can remove the aged rat bone marrow mesenchymal stem cells and play an anti-aging role. Methods: the bone marrow mesenchymal stem cells in the thigh bone marrow cavity of 80 g-100 g male SD rats aged 4 weeks were cultured to the third generation with DMEM / F12 medium containing 10 % fetal bovine serum, D-gal was used to induce bone marrow mesenchymal stem cells to senescence and SA-β-gal staining was used to confirm that the cells were in the state of senescence. The expression levels of SIRT1, SIRT6 and p16, p21 and p53 in the control group and the senescence group were detected by Western blotting. After that, CCK-8 was used to detect the cytotoxicity of linarin and find the target concentration. At last, by Western blotting analysis and immunofluorescence, the expression of aging related proteins in the control group and the treatment group were detected. It was observed whether linarin could clear the aging rat bone marrow mesenchymal stem cells and play an anti-aging role. Results: compared with normal cells, linarin can inhibit the aging of rat bone marrow mesenchymal stem cells, SIRT1, SIRT6 and p16, p21 and p53 of aging MSCs decreased, while the expression of γ - H2AX, a protein related to DNA damage, increased significantly in aging cells, Conclusions: Linarin can scavenge the aged rat bone marrow mesenchymal stem cells in a dose-dependent manner and play an anti-aging role. |
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