摘要
Spexin是一种具有多种生物学功能的神经肽,在代谢疾病及神经系统疾病中发挥重要作用.为了深入研究spexin的生理功能和相关机制,构建了spexin转基因小鼠,通过检测血液中spexin含量以及糖耐受实验,对模型小鼠进行了初步的表型分析.利用piggyBac(PB)转座子技术培育spexin转基因小鼠,采用聚合酶链反应(polymerase chain reaction,PCR)和酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)分别在基因层面和蛋白层面验证了模型动物是否构建成功.之后用饲喂高脂饲料的雄性小鼠进行糖耐受实验,对转基因模型小鼠进行表型分析.PCR结果显示,以靶向spexin的两对引物扩增,转基因型小鼠基因组DNA中能分别扩增出300 bp和301 bp的片段,而野生型小鼠则不能.ELISA结果中,转基因型小鼠血清中spexin含量较野生型小鼠显著升高;同一表型中,雌雄小鼠间血清中spexin水平无明显差异.饲喂高脂饲料85 d内,转基因型小鼠和野生型小鼠体重变化未观察到显著差异;糖耐受结果显示,转基因型小鼠每个时间点血糖浓度均低于野生型小鼠,且在15 min时具有显著性差异.结果显示成功构建了过表达spexin基因的小鼠模型.spexin表达水平的升高在一定程度上提高了C57BL/6小鼠的糖耐受能力.该模型为进一步研究spexin基因在动物体内的功能以及在代谢疾病发病机制中的作用奠定了基础.
Spexin,a neuropeptide with diverse biological functions,plays important roles in metabolic diseases and nervous system diseases.In order to further study the physiological functions of spexin and the underlying mechanisms,we successfully generated transgenic mice with overexpression of spexin.The spexin transgenic mice were generated by using piggyBac(PB)transposon system.The construction of the model animals was verified by PCR and ELISA at the gene level and protein level,respectively.After that,a glucose tolerance test was carried out in male mice fed with high fat diet to analyze the phenotype of transgenic mice.The results of PCR showed that segments with the size of 300 bp and 301 bp were amplified with two pairs of primers targeting spexin from the genomic DNA of spexin transgenic mice but not wild-type C57BL/6 mice.The serum spexin level of transgenic mice was significantly higher than that of the wild-type mice.Meanwhile,there was no significant difference in the serum spexin level between male and female mice in the same phenotype.After 85 days of feeding high-fat diet,there was no significant difference in body weight change between transgenic mice and wild-type mice.The results of glucose tolerance test showed that the blood glucose concentration of transgenic mice at each time point was lower than that of wild-type mice with significant difference at 15 minutes.The results showed that we have successfully generated a mouse model overexpressing spexin gene.Increased expression of spexin can enhance the glucose tolerance ability of C57BL/6 mice to some extent.This model lays a foundation for further study of the function of spexin gene in animals and its role in the pathogenesis of metabolic diseases.
作者
杨春菊
庄敏
赖奇
尹俊林
樊保敏
卞兆祥
林成源
曾广智
YANG Chun-ju;ZHUANG Min;LAI Qi;YIN Jun-lin;FAN Bao-min;BIAN Zhao-xiang;LIN Cheng-yuan;ZENG Guang-zhi(YMU-HKBU Joint Laboratory of Traditional Natural Medicine, Yunnan Minzu University, Kunming 650500, China;School of Chinese Medicine, Hong Kong Baptist University, Hong Kong, China)
出处
《云南民族大学学报（自然科学版）》
CAS
2019年第4期337-343,共7页
Journal of Yunnan Minzu University:Natural Sciences Edition
基金
国家自然科学基金(31660335)
