单细胞海藻提取物对人脐带间充质干细胞的抗衰老作用  

Anti-aging effects of marine phytoplankton on human umbilical cord derived mesenchymal stem cells

作　　者：姚宁[1] 马珊珊[1] 崔渊博[1] 渠瑞娜 王欣欣[2] 孟楠[2] 宋及时 赵云[1] 高军 关方霞[1,2] 

机构地区：[1]郑州大学生命科学学院,郑州450001 [2]郑州大学第一附属医院干细胞研究室,郑州450052 [3]北京高新源新兴技术开发有限公司,北京100029

出　　处：《郑州大学学报（医学版）》2015年第4期458-462,共5页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金资助项目81071008;81471306;U1404313;河南省科技创新人才计划基金资助项目;河南省高校科技创新团队基金资助项目15IRTSTHN02;河南省产学研合作项目142107000008

摘　　要：目的:探讨单细胞海藻(MPPT)提取物对人脐带间充质干细胞(hUC-MSCs)的抗衰老作用。方法:取剖宫产新生儿脐带,分离、传代培养hUC-MSCs,显微镜下观察细胞形态,收集第3、10、15代细胞,qRT-PCR分析不同代细胞中p16、p21、p53、PCNA、Sirt2 mRNA的表达情况。CCK-8法检测不同质量浓度MPPT提取物对第15代hUCMSCs增殖的影响;流式细胞术检测MPPT提取物对hUC-MSCs细胞周期和凋亡的影响;β-半乳糖苷酶染色法检测MPPT提取物对hUC-MSCs衰老的影响;qRT-PCR检测MPPT提取物对hUC-MSCs中p16、p21、p53、PCNA、Sirt2 mRNA表达的影响。结果:第3代细胞呈梭形贴壁细胞,第15代细胞呈不规则状态,易脱壁。随着传代次数的增加,hUC-MSCs中PCNA、Sirt2 mRNA表达减少(F=76.944、25.649,P<0.001),而p16、p21和p53 mRNA表达增加(F=46.714、77.645、845.676,P<0.001);与第15代细胞相比,1 g/L的MPPT提取物可促进第15代hUC-MSCs增殖,促进细胞周期进入S期,抑制细胞凋亡(P均<0.05);MPPT提取物处理组衰老细胞率降低(F=773.557,P<0.001),细胞中PCNA、Sirt2 mRNA表达增加,p16、p21和p53 mRNA表达降低(P均<0.05)。结论:MPPT提取物可能通过调节hUC-MSCs中p16、p21、p53、PCNA和Sirt2等衰老相关基因的表达发挥抗衰老作用。Aim： To explore the anti-aging effects of marine phytoplankton（MPPT） on human umbilical cord derived mesenchymal stem cells（hUC-MSCs）. Methods： hUC-MSCs were isolated from umbilical cords of mature healthy newbo-rns delivered by caesarean section, and then cultured and passaged, while the morphology was observed under microscope. The passage 3, 10, and 15 cells were collected and the mRNA expressions of p16, p21, p53, PCNA, and Sirt2 in different passages of hUC-MSCs were detected by qRT-PCR. CCK-8 assay was performed to detect the effect of MPPT on cell prolif-eration and flow cytometry was used to detect cell cycle and apoptosis in the passage 15 cells. β-galactosidase staining was used to detect the effect of MPPT on cell senescence, and the effect of MPPT on the mRNA expressions of p16, p21, p53, PCNA, and Sirt2 of hUC-MSCs were detected by qRT-PCR. Results： The cell morphology of passage 3 was fusiform and easily adherent, however, the passage 15 hUC-MSCs were flat and poorly adherent. The mRNA expressions of PCNA and Sirt2 reduced gradually along with passaging（F = 76. 944,25. 649,P 〈 0. 001）, while those of p16, p21, and p53 in-creased（F = 46. 714,77. 645,845. 676, P 〈 0. 001）. Compared with the passage 15 cells, 1 g/ L MPPT could promote the proliferation, induce cells into S phase, inhibit apoptosis of the passage 15 hUC-MSCs（P 〈 0. 05）, decrease the positive rate of β-galactosidase positive staining cells（F = 773. 557,P 〈 0. 001）, and increase the mRNA expressions of PCNA and Sirt2,while decrease the mRNA expressions of p16, p21, and p53（P 〈 0. 05）. Conclusion： MPPT has anti-aging effects on hUC-MSCs, and the mechanism is partly through regulating the expressions of p16, p21, p53, PCNA, and Sirt2.

关 键 词：单细胞海藻 人脐带间充质干细胞 衰老 P16 p53 / P21 

分 类 号：Q291[生物学—细胞生物学]