脂肪組織差異表達(dá)基因RETN、CAV1、PLA2G16與阿勒泰羊尾脂沉積代謝關(guān)系的研究
發(fā)布時(shí)間：2018-11-18 09:23
【摘要】：阿勒泰羊是一種脂臀型綿羊品種,主要通過尾部脂肪沉積與代謝消長(zhǎng)規(guī)律適應(yīng)惡劣的環(huán)境以達(dá)到維持機(jī)體正常的新陳代謝和體溫,是研究綿羊尾脂沉積與代謝機(jī)制的理想模型。抵抗素(Resistin,RETN)、小凹蛋白1(Caveolin-1,CAV1)、脂肪特異性磷脂酶A2(Adipose-specific phospholipase A2,PLA2G16)是脂肪細(xì)胞的關(guān)鍵調(diào)控因子,在細(xì)胞分化、增值等方面發(fā)揮了重要的作用。近年來研究發(fā)現(xiàn)RETN、CAV1、PLA2G16均參與脂肪的代謝、分化過程,為了進(jìn)一步證實(shí)上述三個(gè)因子對(duì)綿羊的尾脂沉積與代謝性狀的影響。本論文在模擬阿勒泰羊夏秋季節(jié)脂肪沉積、春冬季節(jié)水草匱乏時(shí)尾脂代謝方式基礎(chǔ)上,利用PCR技術(shù)克隆了綿羊RETN、CAV1、PLA2G16基因,并對(duì)其序列進(jìn)行生物信息學(xué)分析;采用半定量RT-PCR方法檢測(cè)了RETN、CAV1、PLA2G16基因在綿羊主要組織中的表達(dá);同時(shí)利用實(shí)時(shí)熒光定量PCR技術(shù)檢測(cè)了持續(xù)饑餓與充足采食狀態(tài)下阿勒泰羊尾脂RETN、CAV1、PLA2G16基因的差異表達(dá)情況。主要結(jié)果如下:1.以阿勒泰羊?yàn)檠芯繉?duì)象,通過模擬其在夏秋季節(jié)脂肪沉積、春冬季節(jié)水草匱乏時(shí)尾脂的代謝方式,構(gòu)建了饑餓與非饑餓實(shí)驗(yàn)?zāi)Ｐ汀?.以阿勒泰羊尾脂組織c DNA為模板,通過RT-PCR獲得RETN、CAV1、PLA2G16基因完整CDS區(qū)序列。3.半定量RT-PCR檢測(cè)結(jié)果表明,RETN、CAV1、PLA2G16基因均在阿勒泰羊脂肪組織(腸脂和尾脂組織)中呈高豐度表達(dá)。提示RETN、CAV1、PLA2G16基因在阿勒泰羊尾脂生理過程中發(fā)揮重要作用。4.應(yīng)用實(shí)時(shí)熒光定量PCR技術(shù)檢測(cè)了持續(xù)饑餓狀態(tài)下阿勒泰羊尾脂組織RETN、CAV1、PLA2G16基因的表達(dá)變化情況。結(jié)果表明,RETN基因在饑餓試驗(yàn)前后阿勒泰羊尾脂組織中的表達(dá)量存在明顯差異,持續(xù)饑餓(饑餓狀態(tài))阿勒泰羊尾脂組織中RETN的表達(dá)量極顯著高于正常采食狀態(tài)下尾脂組織的表達(dá)量(P0.01);CAV1、PLA2G16基因則結(jié)果相反,正常采食(非饑餓狀態(tài))阿勒泰羊尾脂組織CAV1、PLA2G16的表達(dá)量極顯著高于持續(xù)饑餓狀態(tài)下尾脂組織的表達(dá)量(P0.01)。上述結(jié)果表明,RETN、CAV1、PLA2G16基因在阿勒泰羊尾脂沉積于代謝過程中具有一定的調(diào)控作用,可以作為哺乳動(dòng)物脂肪組織發(fā)育調(diào)控過程重要的候選功能基因。
[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.
【學(xué)位授予單位】：石河子大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S826
本文編號(hào)：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.
【學(xué)位授予單位】：石河子大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S826
【參考文獻(xiàn)】
相關(guān)博士學(xué)位論文 前1條
1 張艷芳;CPT1基因?qū)ωi脂肪沉積的影響及其調(diào)控機(jī)制的研究[D];浙江大學(xué);2010年
，本文編號(hào)：2339628
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