SIRT6基因促进间充质干细胞对犬的抗衰老作用

The SIRT6 gene promotes the anti-aging effects of mesenchymal stem cells in dogs

摘要:

间充质干细胞(mesenchymal stem cells,MSCs)是延缓犬类衰老的有效治疗策略，但是其在体外长期培养时存在易老化、遗传稳定性差等问题，因此探索一种能提高MSCs抗衰老能力的方法具有重要意义。沉默信息调节因子(sirtuin 6,SIRT6)在抗衰方面起着重要作用，本研究构建了过表达SIRT6基因的MSCs，通过姬姆萨染色及衰老相关β-半乳糖苷酶染色发现，SIRT6可以显著提升MSCs的抗衰能力。通过对细胞进行透射电镜拍摄及对氧化应激相关指标的检测发现，SIRT6通过维持线粒体稳态、降低氧化应激水平提高MSCs的抗衰能力。转录组测序分析发现SIRT6主要作用于磷脂酰肌醇3-激酶、丝裂原活化蛋白激酶等衰老及炎症相关通路。利用小鼠及犬衰老模型发现，SIRT6过表达细胞静脉移植后模型小鼠的空间记忆能力显著升高，脏器指数也有明显变化，犬及小鼠血液的抗氧化应激能力提高；SIRT6过表达细胞处理组的脾脏、肝脏形态能够得到有效恢复，其衰老及炎症相关蛋白的表达量显著降低。本研究为SIRT6介导的MSCs抗衰老研究提供了新思路。

Abstract:

Mesenchymal stem cells (MSCs) are an effective therapeutic strategy to delay aging in dogs, they are prone to aging and have poor genetic stability when cultured for a long time in vitro. Therefore, it is of great significance to explore a method to improve the anti-aging ability of MSCs. Previous studies have shown that sirtuin 6 (SIRT6) plays an important role in anti-aging. This study constructed MSCs with overexpressed SIRT6 gene. Through Giemsa staining and senescence-associated β-galactosidase staining, it was found that SIRT6 significantly enhances the anti-aging capacity of MSCs. Transmission electron microscopy imaging and the detection of oxidative stress-related indicators revealed that SIRT6 improves the anti-aging capacity of MSCs by maintaining mitochondrial homeostasis and reducing oxidative stress levels. Transcriptome sequencing analysis revealed that SIRT6 mainly acted on phosphatidylinositol-3-kinase, mitogen-activated protein kinase and other aging and inflammation related pathways. In the establishment and verification of aging models in mice and dogs, it was found that the spatial memory ability of the model mice was significantly increased after intravenous transplantation of SIRT6 overexpression cells, the organ index was also significantly changed, and the anti-oxidative capacity of the dogs and mice blood was improved. The morphology of the spleens and livers in the SIRT6 overexpression cell treatment group could be effectively restored, and the expression levels of aging and inflammation-related proteins were significantly decreased. This study provides a new idea for the study of SIRT6-mediated anti-aging of MSCs.