中国科学院昆明动物研究所机构知识库

人口老龄化是当今社会的一个重要现象，伴随着老年性疾病发病的上升，给个人、家庭和社会带来沉重的负担，如何应对老龄化已经成为了各国面临的共同问题。而长寿人群作为一个健康衰老模型，不仅表现为个体状态较为健康，还可以延缓甚至规避老年性疾病的发病，且这种特性还具有一定的遗传性。这为对抗老龄化提供了一种思路，即以长寿人群为研究对象，探究长寿人群的生存优势及其背后的遗传因素，从而为对抗老年性疾病以及老龄化问题提供参考。有鉴于此，我们对来自海南的长寿家系人群进行了研究。首先，对来自海南省的658个长寿家系外周血样本进行生理指标检测，主要是血常规和血脂血糖，其中包括长寿老人233位，对照个体425位。我们发现长寿老人在一些指标上呈现独有的优势，最为明显的特征是长寿老人在血脂血糖水平上的优势，长寿人群在总胆固醇、低密度脂蛋白胆固醇和血糖等可能引发动脉粥样硬化和心血管疾病的风险因素均随年龄而下降，而具有抵抗和降低这些疾病发病风险的高密度脂蛋白胆固醇在长寿人群中随着年龄上升。除此以为，在血常规指标方面，长寿人群也表现出一些独特优势，比如长寿老人的免疫能力更强，表现为淋巴细胞比例和中性粒细胞比例高于对照且随年龄逐步增加，血红细胞相关功能如血红蛋白浓度、血红细胞比例改变使得他们可以规避一些心血管病的发病，血小板压积在长寿人群中较低。因此推测这些生化指标优势对于长寿人群获得长寿至关重要（第二章）。其次，为了进一步揭示导致这些指标尤其是血脂水平优势的背后原因，在转录组水平对长寿人群进行了研究。首先对转录组差异基因进行分析，发现长寿人群和对照人群之间存在明显的差异基因，而且富集于生物学调控和代谢等多条通路上，比如差异基因SLC16A10本身负责对于甲状腺激素的细胞运输具有重要作用，而血液中甲状腺激素水平在两个群体之间也存在显著差异。接下来，对表达差异基因和生化指标进行了共表达分析，发现差异表达基因对于脂代谢指标变化具有重要贡献。进一步，通过对转录组获得的突变情形进行分析，发现长寿人群在3’UTR上存在较多的突变，结合脂代谢表达差异基因，找到了AQP3和IL15两个基因在3’UTR上呈现突变优势，AQP3对甘油的转运和代谢起重要作用，而IL15对于脂肪组织的脂代谢有重要作用，尤其是影响脂肪酸从头合成的速率。此外，对转录组突变的突变热点进行了研究，发现位于基因MAN1B1和SH3YL1上存在热点差异，这两个基因一方面在长寿和对照群体中存在表达差异，另一方面，在基因的外显子区域存在hotspots差异。MAN1B1基因负责糖基水解酶，该基因表达异常和智力异常有关。SH3YL1基因是磷脂酰肌醇基因，参与G蛋白偶联受体信号通路转导过程。这提示hotspots可能是影响该基因表达差异的原因之一（第三章）。相比于一般衰老，健康长寿人群表现出独特优势，尤其是在老年性疾病发病上，长寿人群往往能够延缓甚至规避老年性疾病的发病。为了探究其背后的遗传学原因，我们选择了常见的老年性疾病阿尔茨海默病和骨质疏松作为研究对象进行研究。首先通过对来自NCBI的疾病芯片数据进行了分析，然后结合长寿人群表达差异基因发现长寿人群和疾病群体的基因表达差异。接下来，采取DNA检测的办法进行了进一步研究。为了研究长寿人群阿尔茨海默病上的低发病率原因，对负责β淀粉样蛋白编码的基因APP基因进行了研究，在海南和四川两独立长寿群体样本中分别采取sanger测序和HRM的方法研究了APP基因上的保护性突变A673T，随后在海南人群中对负责β淀粉样蛋白编码的APP16和17外显子进行了sanger测序检测，最后通过minigene的办法对APP基因15内含子上的两个长寿特有突变进行验证。但是并没有发现相应的保护机制，结合中国人群阿尔茨海默病发病率低于欧美人群，这提示可能存在其他保护机制。此外，和日本科学家一道对和成骨相关的基因编码四联蛋白编码CLEC3B上的错义突变p.S106G进行了研究，通过对953例来自海南的个体进行研究（其中长寿448位，对照490位），发现该突变在中国长寿人群和对照人群中呈现显著差异（p=0.047），结合日本科学家的研究，证实这是一个可能对东亚广泛存在的长寿关联突变（（第四章）。综上，本研究揭示：1）长寿老人在血液细胞和血脂血糖上存在生存优势 2）长寿人群基因表达差异及基因上的突变可能是导致长寿老人血脂优势的重要因素 3）长寿人群在对抗阿尔茨海默病上可能存在其他机制 ，而CLEC3B基因上的显著突变p.S106G可能对减轻骨质疏松具有一定的效果。

The aging of the population is an important phenomenon in the current society. As the physical function of the aging population declines, and the incidence of senile diseases increases, it places a heavy burden on individuals, families and society. The problem of aging has become a problem faced by all countries. However, the longevity population, with a distinct trend, not only shows relatively healthy individual state, but also can delay or even evade the onset of senile diseases, and this characteristic also has a certain heredity. This provides a way to combat ageing, that is, taking the longevity population as the research object, exploring the survival advantages of longevity groups and the genetic factors behind them, so as to provide reference for combating age-related diseases and aging.In view of this, we studied the longevity families from Hainan.First, the physiological indicators of peripheral blood samples from 658 longevity families from Hainan Province were examined, mainly blood routine and blood lipids, including 233 elderly longevity and 425 control individuals. We have found the longevities have unique advantages in some indicators. The most obvious feature is the advantage in blood-lipid and glucose levels. Risk factors for atherosclerosis and cardiovascular disease such as total cholesterol, low-density lipoprotein cholesterol, and blood glucose all decline with age, while high-density lipoprotein cholesterol, which has the ability to resist and reduce the risk of developing these diseases, rises with age in long-lived people. In addition to this, in terms of blood routine indicators, the longevity population also showed some unique advantages. For example, the longevities had stronger immunity, manifested as a higher ratio of lymphocytes and neutrophils than control and gradually increased with age; red blood cells such as hemoglobin concentration , the proportion of red blood cells and platelet pressure also changed in long-lived people which may allow them to avoid the onset of some cardiovascular diseases. Therefore, it is inferred that the advantages of these biochemical indicators may be crucial for longevity groups in achieving healthy longevity (Chapter 2).Second, in order to further reveal the underlying causes of these indicators, especially the level of blood lipids, longevity populations were studied at the transcriptome level. Firstly, we analyzed the transcriptome differential expressed genes, and found that there were significant differences between longevity and control populations, and they were enriched in multiple pathways such as biological regulation and metabolism. For example, the differential gene SLC16A10 is responsible for cell transport of thyroid hormones. Thyroid hormones levels are significant differences in blood thyroid hormone levels between the two groups. Next, co-expression analysis of differentially expressed genes and biochemical markers revealed that differentially expressed genes have important contributions to changes in lipid metabolism. Further, by analyzing the mutations obtained in the transcriptome, it was found that there were more mutations in the longevity group on the 3'UTR, together with genes that were differentially expressed in lipid metabolism, AQP3 and IL15 were found exhibiting mutational superiority in the 3' UTR. AQP3 plays an important role in the transport and metabolism of glycerol, while IL15 plays an important role in the lipid metabolism of adipose tissue, especially the rate of de novo fatty acid synthesis. In addition, the hot spots of mutations in transcriptomes were studied and found that there are hotspot differences in the genes MAN1B1 and SH3YL1. On the one hand, these two genes have differences in expression in longevity and control populations, and on the other hand, exons in genes. There are hotspots differences in the area. The MAN1B1 gene is responsible for glycosyl hydrolase, and abnormal expression of this gene is associated with abnormal intelligence. The SH3YL1 gene is a phosphatidylinositol gene involved in the transduction of G protein-coupled receptor signaling pathways. This suggests that hotspots may be one of the reasons that affect the gene expression differences (Chapter 3).Compared with general aging, people with healthy longevity show unique advantages, especially in the pathogenesis of senile diseases, longevity people can often delay or even avoid the onset of senile diseases. In order to explore the genetic reasons behind it, we chose the common senile disease Alzheimer's disease and osteoporosis as research objects. Firstly, we analyzed the data of disease microarrays from NCBI, and then combined the expression of differential genes in longevity populations to find differences in gene expression between longevity and disease groups. Next, the DNA testing method was used for further study. In order to study the causes of low incidence of Alzheimer's disease in longevity populations, the APP gene responsible for β-amyloid protein was studied, and sanger sequencing and HRM methods were used in the samples of two independent longevity groups in Hainan and Sichuan respectively. A protective mutation A673T on the APP gene, followed by sanger sequencing of the APP16 and 17 exons encoding beta amyloid in the Hainan population, and finally through the minigene approach to the intron 15 of the APP gene 15 was verified. However, no corresponding protection mechanism has been found. The incidence of Alzheimer's disease combined with Chinese population is lower than that of European and American populations, suggesting that there may be other protective mechanisms. In addition, we work together with the Japanese scientists in studing the CLEC3B missense mutation p.S106G 953 individuals from Hainan (including 448 longevity and 490 controls). The mutation was found to be significantly different between Chinese longevity and control populations (p=0.047). Combined with Japanese scientists' studies, it was confirmed that this mutation is a long-lived associated mutation that may be widespread in East Asia (Chapter 4).In summary, this study reveals: 1) longevities have survival advantages in blood cells and blood lipids and blood glucose; 2) longevities have have significant differences in gene expression and genetic mutations. 3)There may be other mechanisms on Alzheimer's disease, and the significant mutation p.S106G on the CLEC3B gene may have a certain effect on reducing osteoporosis.