糖尿病研究的蝙蝠动物模型和SLC2A2基因研究

发布时间：2018-01-09 19:27

本文关键词：糖尿病研究的蝙蝠动物模型和SLC2A2基因研究　出处：《华东师范大学》2016年博士论文　论文类型：学位论文

【摘要】：蝙蝠属于哺乳动物纲翼手目,物种数量占哺乳动物的五分之一。食果蝙蝠较其它蝙蝠具有更好的维持糖稳态、胰岛素敏感性和胰岛素降血糖能力,食虫蝙蝠有在冬眠前期快速增加体重、储存大量脂肪和较强的白色脂肪棕样化能力,因此蝙蝠可以成为人类糖尿病研究和治疗中非常好的动物模型。身体质量指数和血糖是常用的糖尿病研究方法。为了建立糖尿病研究的蝙蝠模型,本文探索并建立了蝙蝠的身体质量指数(body mass index,BMI)和前臂长质量指数(forearm mass index,FMI),并对这两个指数与血糖水平的相关性、蝙蝠在急性糖耐量实验中的表现及其分子机制进行了研究。本研究建立了149种蝙蝠的BMI指数和860种蝙蝠的FMI指数。相比较而言,食虫蝙蝠拥有更低的BMI和FMI,而食果蝙蝠的BMI和FMI更高,杂食蝙蝠、吸血蝙蝠和食肉蝙蝠的BMI和FMI介于食虫蝙蝠和食果蝙蝠之间,反映了食果蝙蝠比食虫蝙蝠有更高的正常体重基准。通过比较,本文发现人类BMI标准不适合蝙蝠,为此需要建立蝙蝠自身超重和肥胖的判断标准,通过对杂食蝙蝠在以昆虫或水果为主要,甚至唯一食物来源时,其BMI和FMI两指数和血糖相关性以及葡萄糖耐量实验将有助于该标准的确立。随后对BMI和FMI指数与蝙蝠血糖的种间相关性研究,发现BMI和FMI指数与血糖有种间水平的负相关性,即拥有较高BMI和FMI值的蝙蝠有较低的血糖水平,而拥有较低BMI和FMI的蝙蝠血糖较高,这与种内的BMI与血糖水平的正相关性不同。针对这种特别的现象,本文又对蝙蝠的急性糖耐量实验进行了研究,在对食果的棕果蝠和食虫的大蹄蝠的2克葡萄糖耐量实验中,棕果蝠表现出了更低的空腹血糖、更快速地到达血糖峰值,更快速地恢复血糖水平到正常范围和更小的血糖曲线下面积的特点,反映了食果蝙蝠比食虫蝙蝠更强的维持糖稳态的能力和胰岛素降低血糖的能力。血糖可以通过葡萄糖运输蛋白(GLUT)被分别吸收进入肝脏、肌肉和脂肪组织,从而降低血糖实现维持糖稳态的能力。SLC2A2基因的编码蛋白(GLUT2)是参与调控葡萄糖进出肝脏的关键蛋白。为了研究其在蝙蝠维持糖稳态和胰岛素降低血糖过程中的作用,本文进一步对蝙蝠SLC2A2基因在分子进化、转录和翻译水平的表达进行了研究。其中,对蝙蝠SLC2A2基因的分子进化研究发现,无论是食果蝙蝠、食虫蝙蝠,还是其各自的祖先进化枝,都未检测到正选择作用信号。然而,在SLC2A2基因启动子区,所有检测的食果蝙蝠都缺失了一段长度为11 bp的核苷酸序列,而该段序列被预测为转录抑制因子ZNF354C的识别结合位点。随后的荧光定量PCR和蛋白质杂交实验证实缺失该段序列的食果蝙蝠SLC2A2基因在转录和翻译水平,都比仍然存在该段序列的食虫蝙蝠有更高的表达水平,这可能导致了食果蝙蝠和食虫蝙蝠糖代谢能力的差别。
[Abstract]:The bat belongs to mammalia Chiroptera species, which accounts for 1/5 of all mammals. Fruit bats than other bats has better maintain glucose homeostasis, insulin sensitivity and insulin hypoglycemic ability, there is a rapid increase in weight in insectivorous bats dormancy, storing large amounts of fat and strong white fat brown like ability, it can become very bat animal model good study and treatment of human diabetes. Body mass index and blood glucose of diabetes is the common research methods. In order to bat model in diabetes research, this paper explores and establishes a body mass index of bats (body mass index, BMI) and forearm length (forearm mass index quality index, FMI), and the correlation of this two index and blood glucose levels, bats in acute glucose tolerance and its molecular mechanism in experimental performance were studied. This study established 149 The bat's BMI index and FMI index of the 860 species of bats. In comparison, insectivorous bats have lower BMI and FMI, while the fruit bat BMI and higher FMI, between the vampire bat and bat omnivorous, carnivorous bat BMI and FMI between insectivorous bats and fruit bats, reflects the fruit eating bat has normal weight higher than the benchmark insectivorous bats. By comparison, this paper found that the human BMI standard is not suitable for bats, this requires the establishment of standards and their fat bat overweight, the bats in omnivorous insects or fruit as the main, or even the only source of food, the BMI and the FMI two index and the correlation between blood glucose and glucose tolerance the establishment of experimental will contribute to this standard. Then the BMI and FMI index and bat blood glucose interspecific correlation study found negative correlation between BMI and FMI index and blood glucose levels between species, which has high BMI and FMI value of the bat There is a low glucose level, while BMI and FMI have lower bat high blood sugar, and this kind of different positive correlation in BMI and the level of blood glucose. In view of this special phenomenon, the acute glucose tolerance test of bats were studied on fruits and large insectivorous bats Hipposideros 2 grams of glucose tolerance test, showed lower Rousettus fasting blood glucose, blood glucose peak arrived more quickly, more quickly to restore blood glucose levels to normal blood glucose curve area and a smaller range of fruit bats reflects stronger than insectivorous bats maintain glucose homeostasis and ability insulin's ability to lower blood sugar. Blood glucose by glucose transport protein (GLUT) were absorbed into the liver, muscle and adipose tissue, thereby reducing the ability of.SLC2A2 gene can maintain glucose homeostasis of blood glucose (GLUT2) encoding protein is involved in the regulation of Key protein import glucose in the liver. In order to study the maintenance of glucose homeostasis and insulin in bat glucose lowering effect in process, in order to bat SLC2A2 gene in molecular evolution, the expression levels of transcription and translation were studied. Among them, the study of molecular evolution of SLC2A2 gene in bats found either frugivorous bats, insectivorous bats still, their ancestral clade, not detected positive selection signal. However, in the promoter region of SLC2A2 gene, all detected fruit bats are missing nucleotide sequence with a length of 11 BP, and the sequence was predicted to identify transcriptional repressor ZNF354C binding sites. Then the fluorescence quantitative PCR and Western blot experiments confirmed that deletion of the sequence of the frugivorous bat SLC2A2 gene on the transcription and translation level than the sequences of insectivorous bats still have higher The expression level, which may lead to the difference between the sugar metabolism of the fruit bats and the insectivorous bats.

【学位授予单位】：华东师范大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R587.1;R-332

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