蒙花苷消除衰老骨髓间充质干细胞的衰老表型发挥抗衰老作用

Linarin Play an Anti-aging Role by Eliminates the Senescent Phenotype of Bone Marrow Mesenchymal Stem Cells

目的:探讨蒙花苷(Linarin,LR)是否可以消除衰老骨髓间充质干细胞(BMSC)的衰老表型发挥抗衰老作用。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诱导骨髓间充质干细胞衰老,使用SA-β-gal染色、活性氧检测、蛋白免疫印迹验证骨髓间充质干细胞细胞衰老状态,通过蛋白免疫印迹检测衰老BMSC与正常BMSC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,我们通过不同浓度LR处理衰老骨髓间充质干细胞。最后,通过蛋白印迹分析检测未处理组与LR处理组衰老相关蛋白表达情况。观察蒙花苷能否可以消除衰老的大鼠骨髓间充质干细胞衰老表型。结果: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消除衰老的大鼠骨髓间充质干细胞的衰老表型发挥抗衰老作用。

Objective:To investigate whether linarin(LR)can eliminate the aging phenotype of aging bone marrow mesenchymal stem cells(BMSC)and play an anti-aging role.Methods:The bone marrow mesenchymal stem cells(BMSCs)in the thigh marrow cavity of male SD rats aged from 80 g to 100 g were used%DMEM/F12 medium of fetal bovine serum was cultured to the third generation.D-gal was used to induce the aging of BMSCs.SA-β-gal staining,active oxygen detection and Western blotting were used to verify the aging state of BMSCs.The expression of SIRT1,SIRT6 and p16,p21 and p53 were detected by western blotting Level.After that,we treated the aged BMSCs with different concentrations of LR.Finally,Western blot analysis was used to detect the expression of aging related proteins in the untreated and LR treated groups.To observe whether Linarin can eliminate the aging phenotype of bone marrow mesenchymal stem cells in aging BMSC.Results:In senescent cells,SIRT1 and SIRT6 decreased,p16,p21 and p53 increased,and the expression ofγ-H2 AX related to DNA damage increased significantly.However,after LR treatment,p16,p21,p53 andγ-H2 AX protein positive cells were significantly decreased.Conclusions:Linarin can eliminate the aging phenotype of aging bone marrow mesenchymal stem cells in a dose-dependent manner and play an anti-aging role.