恒河猴川西亚种的Ⅱ型糖尿病遗传特征分析

发布时间：2018-03-17 10:55

本文选题：恒河猴川西亚种　切入点：II型糖尿病　出处：《四川抗菌素工业研究所》2015年硕士论文　论文类型：学位论文

【摘要】：II型糖尿病（T2DM）是环境和遗传共同作用的结果，家族史、肥胖、年龄、高脂饮食、缺乏运动和压力等均为危险因素。研究T2DM的发病机制以及潜在的治疗手段，需要合适的T2DM动物模型。目前使用最广泛的是啮齿类动物T2DM模型，但是由于其与人亲缘关系远，机体生理特征差别大，因此作为模型研究人类疾病并不能很准确模拟患者的状况。非人灵长类动物由于在种属关系上与人接近，被认为是最接近人类的疾病研究模型动物。已有的报道提示，恒河猴自发T2DM发病情况与人十分相似，疾病的发生发展过程及临床表现与人T2DM相似。但是，恒河猴自发T2DM的遗传因素研究未见报道。我们采用静脉葡萄糖耐量试验、BMI、空腹血糖、空腹胰岛素以及糖化血红蛋白等指标对50只在相同环境条件下饲养的恒河猴进行监测，发现了8只T2DM恒河猴。之后，采用微卫星分子标记法（SSR）对发生糖尿病和未发生糖尿病的恒河猴进行了亲缘关系分析，，同时采用直接测序法研究了Genbank中的恒河猴单核苷酸多态性（SNP）位点KCNJ11（rs196622309）、HHEX/IDE（rs195115827，rs194506087）、ADIPOQ（rs196717151，rs196591545，rs196142463，rs196060906，rs195222188）的基因多态性与疾病发生的关联性。结果显示，糖尿病恒河猴个体之间的亲缘关系较近，与不易感恒河猴之间的亲缘关系较远。8个SNP位点与关联性分析结果表明，ADIPOQ（rs196717151）、ADIPOQ（rs195222188）、HHEX/IDE（rs195115827）、ADIPOQ（rs196060906）4个位点的聚合酶链式反应在易感糖尿病的恒河猴中不能得到目的条带，故未进行下一步的测序分析。KCNJ11（rs196622309）、HHEX/IDE （rs194506087）、ADIPOQ（rs196142463）3个SNP位点在T2DM和不易感T2DM恒河猴中均只出现了同一种基因型，没有出现多态性。ADIPOQ（rs196591545）在T2DM和不易感T2DM恒河猴中都出现了GG、GC、CC三种基因型，表现出多态性。四个位点的序列都与Genbank中的序列一致。rs196591545C/G位点的基因型频率和等位基因频率，在T2DM和不易感T2DM恒河猴中分别为GG0.375/0.1428、GC0.375/0.7143、 CC0.25/0.1428、 G0.5625/0.5、 C0.4375/0.5。此外，在rs195522309位点下游发现了一个新的SNP，在T2DM和不易感T2DM恒河猴中都表现出多态性，产生CC、GC、GG三种基因型。易感糖尿病恒河猴的亲缘关系较近，可能有共同的遗传特征。但由于样本量太少，还没有发现SNP与恒河猴T2DM的关联性。
[Abstract]:Type II diabetes mellitus (T2DM) is the result of environmental and genetic interaction. Family history, obesity, age, high-fat diet, lack of exercise and stress are all risk factors. A suitable animal model of T2DM is needed. At present, the rodent model of T2DM is widely used. Therefore, the study of human diseases as a model can not accurately simulate the condition of patients. Non-human primates are considered to be the closest model animals to human disease because they are closely related to human species. The incidence of spontaneous T2DM in rhesus monkey is very similar to that in human, and the process and clinical manifestation of the disease are similar to that of human T2DM. However, the genetic factors of spontaneous T2DM in rhesus monkey have not been reported. We monitored 50 rhesus monkeys fed under the same environment with BMI, fasting blood glucose, fasting insulin and glycosylated hemoglobin, and found 8 T2DM rhesus monkeys. The genetic relationship of rhesus monkeys with and without diabetes mellitus was analyzed by microsatellite molecular marker method (SSRs). At the same time, direct sequencing was used to study the association between the genetic polymorphisms of Rhesus Monkey single nucleotide Polymorphic SNPs (KCNJ11622309) locus KCNJ11622309, rs195115827rs194506087rs194506087rs196717151rs196591545rs196142463rs196060906rs195222188). The results showed that the genetic relationship between individuals of diabetic rhesus monkey was close. The results of 8 SNP loci and association analysis showed that the four loci of ADIPOQN rs1952221888 HHEX / IDEN rs195115827) could not be detected in the rhesus monkey susceptible to diabetes mellitus by polymerase chain reaction (PCR) at 4 loci of the rhesus monkey, which was susceptible to diabetes mellitus (Rhesus mulatta), and the results showed that there were no bands of polymerase chain reaction at the 4 loci of ADIPOQN rs196060906 in the rhesus monkey susceptible to diabetes mellitus (Rhesus mulatta). Therefore, no further sequencing analysis was carried out. KCNJ11rs196622309 (HHEX / Ide rs194506087rs194506087rs196142463) three SNP loci had only the same genotype in T2DM and non-susceptible T2DM rhesus monkeys, and no polymorphism. ADIPOQRs196591545) in T2DM and non-susceptible T2DM rhesus monkeys, there were three genotypes of GGGCCC in both T2DM and non-susceptible T2DM rhesus monkeys. The sequence of the four loci is consistent with the sequence of Genbank. The genotype frequency and allelic frequency of the locus G in T2DM and non-susceptible T2DM rhesus monkeys are G0.375 / 0.1428 GC0.3750.7143, CC0.25 / 0.1428, G0.5625- / 0.5, C0.4375- / 0.5respectively. A new SNPs was found downstream of the rs195522309 locus, which showed polymorphism in both T2DM and non-susceptible T2DM rhesus monkeys, and produced three genotypes of CCGCG-GG. The genetic relationship of susceptible diabetic rhesus monkeys is close, which may have the same genetic characteristics. However, because of the small sample size, no association between SNP and Rhesus monkey T2DM has been found.
【学位授予单位】：四川抗菌素工业研究所
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R587.1

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