sirt2基因高表达对人脐带间充质干细胞衰老的抑制作用
Inhibition of sirt2 overexpression on aging of human umbilical cord mesenchymal stem cells

目的：观察sirt2基因高表达对人脐带间充质干细胞（hUC？？MSCs）衰老的影响。方法：扩增人sirt2 ORF序列，利用腺病毒载体构建sirt2基因高表达的重组腺病毒（pAd？？sirt2），空斑法测定病毒滴度；感染hUC？？MSCs，倒置荧光显微镜下观察荧光表达情况；实验分为P3组（第3代hUC？？MSCs）、P15组（第15代hUC？？MSCs）和感染组（用pAd？？sirt2腺病毒感染第15代hUC？？MSCs），CCK？？8检测细胞增殖情况，流式细胞术分析细胞周期变化，β？舶肴樘擒彰溉旧？后检测阳性细胞率。结果：成功制备pAd？？sirt2重组腺病毒，病毒滴度为1.8×1011 pfu/L；荧光观察证实Sirt2蛋白在hUC？？MSCs中有效表达；与P15组相比，sirt2高表达可促进hUC？？MSCs增殖，并将细胞周期阻滞在S期，β？舶肴樘擒彰秆粜韵赴？率降低(P均＜0.05)。结论：sirt2基因高表达可促进hUC？？MSCs增殖，延长细胞周期，并有效抑制hUC？？MSCs的衰老。
Aim: To construct a recombinant adenovirus containing sirt2 gene and to explore the impact of sirt2 overexpression on the aging of human umbilical cord mesenchymal stem cells(hUC？？MSCs). Methods: Human sirt2 ORF was generated by RT？？PCR and the recombinant adenovirus(pAd？？sirt2) was constructed.The viral titer was determined by air？？dilution method.After being transfected into hUC？？MSCs, the green fluorescence was observed under fluorescence microscope. hUC？？MSCs were allocated into P3 group(P3 hUC？？MSCs), P15 group(P15 hUC？？MSCs) and transfected group(transfected hUC？？MSCs by pAd？？sirt2). CCK？？8 assay was performed to detect the proliferation of hUC？？MSCs and flow cytometry, to analyze the cell cycle. SA？拨陋？Gal was used to detect the cell senescence. Results: The recombinant adenovirus was constructed and the titer was 1.8×1011 pfu/L. After infection by pAd？？sirt2, P15 hUC？？MSCs expressed green fluorescence protein clearly under the fluorescence microscope. Compared with P15 group, sirt2 overexpression could promote the cell proliferation, induced more cells at S phase,and decrease the positive rate of SA？拨陋？Gal(P＜0.05). Conclusion: sirt2 overexpression could promote hUC？？MSCs proliferation,extend cell cycle,and inhibit cell senescence