脂肪组织差异表达基因RETN、CAV1、PLA2G16与阿勒泰羊尾脂沉积代谢关系的研究
发布时间：2018-11-18 09:23
【摘要】：阿勒泰羊是一种脂臀型绵羊品种,主要通过尾部脂肪沉积与代谢消长规律适应恶劣的环境以达到维持机体正常的新陈代谢和体温,是研究绵羊尾脂沉积与代谢机制的理想模型。抵抗素(Resistin,RETN)、小凹蛋白1(Caveolin-1,CAV1)、脂肪特异性磷脂酶A2(Adipose-specific phospholipase A2,PLA2G16)是脂肪细胞的关键调控因子,在细胞分化、增值等方面发挥了重要的作用。近年来研究发现RETN、CAV1、PLA2G16均参与脂肪的代谢、分化过程,为了进一步证实上述三个因子对绵羊的尾脂沉积与代谢性状的影响。本论文在模拟阿勒泰羊夏秋季节脂肪沉积、春冬季节水草匮乏时尾脂代谢方式基础上,利用PCR技术克隆了绵羊RETN、CAV1、PLA2G16基因,并对其序列进行生物信息学分析;采用半定量RT-PCR方法检测了RETN、CAV1、PLA2G16基因在绵羊主要组织中的表达;同时利用实时荧光定量PCR技术检测了持续饥饿与充足采食状态下阿勒泰羊尾脂RETN、CAV1、PLA2G16基因的差异表达情况。主要结果如下:1.以阿勒泰羊为研究对象,通过模拟其在夏秋季节脂肪沉积、春冬季节水草匮乏时尾脂的代谢方式,构建了饥饿与非饥饿实验模型。2.以阿勒泰羊尾脂组织c DNA为模板,通过RT-PCR获得RETN、CAV1、PLA2G16基因完整CDS区序列。3.半定量RT-PCR检测结果表明,RETN、CAV1、PLA2G16基因均在阿勒泰羊脂肪组织(肠脂和尾脂组织)中呈高丰度表达。提示RETN、CAV1、PLA2G16基因在阿勒泰羊尾脂生理过程中发挥重要作用。4.应用实时荧光定量PCR技术检测了持续饥饿状态下阿勒泰羊尾脂组织RETN、CAV1、PLA2G16基因的表达变化情况。结果表明,RETN基因在饥饿试验前后阿勒泰羊尾脂组织中的表达量存在明显差异,持续饥饿(饥饿状态)阿勒泰羊尾脂组织中RETN的表达量极显著高于正常采食状态下尾脂组织的表达量(P0.01);CAV1、PLA2G16基因则结果相反,正常采食(非饥饿状态)阿勒泰羊尾脂组织CAV1、PLA2G16的表达量极显著高于持续饥饿状态下尾脂组织的表达量(P0.01)。上述结果表明,RETN、CAV1、PLA2G16基因在阿勒泰羊尾脂沉积于代谢过程中具有一定的调控作用,可以作为哺乳动物脂肪组织发育调控过程重要的候选功能基因。
[Abstract]:Altay Sheep is a kind of fat-buttock sheep breed. It is an ideal model to study the mechanism of fat deposition and metabolism in sheep tail, which mainly adapts to the bad environment through the law of fat deposition and metabolism in the tail to maintain the normal metabolism and body temperature of the body. Resistin (Resistin,RETN), concave protein 1 (Caveolin-1,CAV1) and fatty specific phospholipase A2 (Adipose-specific phospholipase A2) are the key regulatory factors of adipocytes, which play an important role in cell differentiation and proliferation. In recent years, it has been found that RETN,CAV1,PLA2G16 is involved in fat metabolism and differentiation. In order to further confirm the effects of these three factors on tail fat deposition and metabolic traits in sheep. On the basis of simulating fat deposition in summer and autumn season and tail fat metabolism mode in spring and winter, sheep RETN,CAV1,PLA2G16 gene was cloned by PCR technique and its sequence was analyzed by bioinformatics. Semi-quantitative RT-PCR was used to detect the expression of RETN,CAV1,PLA2G16 gene in sheep tissues. At the same time, the differential expression of RETN,CAV1,PLA2G16 gene in Altay's tail fat was detected by real-time fluorescence quantitative PCR. The main results are as follows: 1. The experiment model of starvation and non-starvation was established by simulating fat deposition in summer and autumn and tail fat metabolism in spring and winter. The complete CDS region of RETN,CAV1,PLA2G16 gene was obtained by RT-PCR using c DNA from Altay's tail fat tissue as a template. The results of semi-quantitative RT-PCR showed that RETN,CAV1,PLA2G16 gene was highly expressed in fat tissues of Altay sheep (intestinal fat and tail fat). It is suggested that RETN,CAV1,PLA2G16 gene plays an important role in the physiological process of tail fat in Altay. 4. The changes of RETN,CAV1,PLA2G16 gene expression in the tail fat tissue of Altay were detected by real-time fluorescence quantitative PCR. The results showed that there was significant difference in the expression of RETN gene in the tail fat of Altay sheep before and after starvation test. The expression of RETN in the tail fat of Altay was significantly higher than that in the normal feeding (P0.01). On the contrary, the expression of CAV1,PLA2G16 in normal (non-hungry) Altay tail fat tissues was significantly higher than that in persistent starvation (P0.01). These results suggest that RETN,CAV1,PLA2G16 gene may play an important role in the regulation of fat deposition in Altay and may be an important candidate gene for the regulation of adipose tissue development in mammals.
【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S826
本文编号：2339628
[Abstract]:Altay Sheep is a kind of fat-buttock sheep breed. It is an ideal model to study the mechanism of fat deposition and metabolism in sheep tail, which mainly adapts to the bad environment through the law of fat deposition and metabolism in the tail to maintain the normal metabolism and body temperature of the body. Resistin (Resistin,RETN), concave protein 1 (Caveolin-1,CAV1) and fatty specific phospholipase A2 (Adipose-specific phospholipase A2) are the key regulatory factors of adipocytes, which play an important role in cell differentiation and proliferation. In recent years, it has been found that RETN,CAV1,PLA2G16 is involved in fat metabolism and differentiation. In order to further confirm the effects of these three factors on tail fat deposition and metabolic traits in sheep. On the basis of simulating fat deposition in summer and autumn season and tail fat metabolism mode in spring and winter, sheep RETN,CAV1,PLA2G16 gene was cloned by PCR technique and its sequence was analyzed by bioinformatics. Semi-quantitative RT-PCR was used to detect the expression of RETN,CAV1,PLA2G16 gene in sheep tissues. At the same time, the differential expression of RETN,CAV1,PLA2G16 gene in Altay's tail fat was detected by real-time fluorescence quantitative PCR. The main results are as follows: 1. The experiment model of starvation and non-starvation was established by simulating fat deposition in summer and autumn and tail fat metabolism in spring and winter. The complete CDS region of RETN,CAV1,PLA2G16 gene was obtained by RT-PCR using c DNA from Altay's tail fat tissue as a template. The results of semi-quantitative RT-PCR showed that RETN,CAV1,PLA2G16 gene was highly expressed in fat tissues of Altay sheep (intestinal fat and tail fat). It is suggested that RETN,CAV1,PLA2G16 gene plays an important role in the physiological process of tail fat in Altay. 4. The changes of RETN,CAV1,PLA2G16 gene expression in the tail fat tissue of Altay were detected by real-time fluorescence quantitative PCR. The results showed that there was significant difference in the expression of RETN gene in the tail fat of Altay sheep before and after starvation test. The expression of RETN in the tail fat of Altay was significantly higher than that in the normal feeding (P0.01). On the contrary, the expression of CAV1,PLA2G16 in normal (non-hungry) Altay tail fat tissues was significantly higher than that in persistent starvation (P0.01). These results suggest that RETN,CAV1,PLA2G16 gene may play an important role in the regulation of fat deposition in Altay and may be an important candidate gene for the regulation of adipose tissue development in mammals.
【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S826
【参考文献】
相关博士学位论文 前1条
1 张艳芳;CPT1基因对猪脂肪沉积的影响及其调控机制的研究[D];浙江大学;2010年
，本文编号：2339628
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