本文選題：能量負(fù)平衡　切入點(diǎn)：脂肪細(xì)胞　出處：《吉林大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：圍產(chǎn)期是奶牛酮病和脂肪肝等能量代謝障礙性疾病的高發(fā)期,由于奶牛干物質(zhì)攝入減少而能量需求增加導(dǎo)致能量負(fù)平衡,進(jìn)而引起脂肪大量動(dòng)員,造成酮病、脂肪肝等能量代謝障礙性疾病。所以能量負(fù)平衡所引起的脂肪動(dòng)員是奶牛圍產(chǎn)期能量代謝障礙性疾病的重要環(huán)節(jié)。此外,脂肪組織作為機(jī)體重要的內(nèi)分泌器官,分泌多種細(xì)胞因子如腫瘤壞死因子(TNFα)、白介素1β(IL-1β)和白介素6(IL-6)等,而這些細(xì)胞因子與酮病和脂肪肝等能量代謝障礙性疾病及乳房炎和子宮內(nèi)膜炎等感染性疾病的發(fā)生發(fā)展密切相關(guān)。PLIN1是在脂肪細(xì)胞中大量表達(dá)的一種脂滴包被蛋白,在脂代謝過(guò)程中起著關(guān)鍵的作用。明確圍產(chǎn)期能量負(fù)平衡奶牛脂肪組織的脂代謝特征,特別是脂代謝酶表達(dá)和活性的改變是緩解脂肪動(dòng)員,防治圍產(chǎn)期能量代謝障礙性疾病的關(guān)鍵。同時(shí),PLIN1對(duì)脂肪細(xì)胞炎性因子的表達(dá)分泌可能也有著特別的作用。因此,本題擬通過(guò)圍產(chǎn)期病牛在體實(shí)驗(yàn)和原代培養(yǎng)脂肪細(xì)胞過(guò)表達(dá)及沉默PLIN1體外實(shí)驗(yàn),以期揭示PLIN1對(duì)脂肪組織脂代謝的調(diào)節(jié)機(jī)制以及炎性反應(yīng)的發(fā)生機(jī)制,為采用緩解脂肪動(dòng)員防治奶牛圍產(chǎn)期能量負(fù)平衡性疾病奠定理論基礎(chǔ)。體外分離培養(yǎng)奶牛原代脂肪細(xì)胞,轉(zhuǎn)染PLIN1過(guò)表達(dá)腺病毒和沉默si RNA。結(jié)果表明PLIN1能顯著上調(diào)SREBP-1c m RNA和蛋白表達(dá)并增強(qiáng)其下游脂肪酸合成關(guān)鍵酶ACC、FAS、SCD以及DGAT1和DGAT2的表達(dá),同時(shí)下調(diào)PPARγm RNA和蛋白表達(dá),且抑制脂肪分解關(guān)鍵酶HSL和ATGL的表達(dá),最終導(dǎo)致脂肪細(xì)胞中脂質(zhì)的積累和脂滴的增大;沉默PLIN1能顯著下調(diào)SREBP-1c m RNA和蛋白表達(dá)并抑制其下游脂肪酸合成關(guān)鍵酶ACC、FAS、SCD以及DGAT1和DGAT2的表達(dá),同時(shí)上調(diào)PPARγm RNA和蛋白表達(dá),且促進(jìn)脂肪分解關(guān)鍵酶HSL和ATGL的表達(dá),最終導(dǎo)致脂肪細(xì)胞中脂質(zhì)的消耗和脂滴的減少。體外分離培養(yǎng)奶牛原代脂肪細(xì)胞,轉(zhuǎn)染PLIN1過(guò)表達(dá)腺病毒和沉默si RNA并添加LPS刺激。結(jié)果表明,過(guò)表達(dá)PLIN1可以抑制NF-κB炎癥信號(hào)通路的激活,使TNFα、IL-1β和IL-6等炎性因子的表達(dá)量下降,起到抗炎作用;沉默PLIN1后,NF-κB炎癥信號(hào)通路的激活增強(qiáng),TNFα、IL-1β和IL-6等炎性因子的表達(dá)量顯著增加,促進(jìn)炎癥反應(yīng)的發(fā)生及發(fā)展。以上體外實(shí)驗(yàn)結(jié)果表明,PLIN1促進(jìn)TG在脂肪細(xì)胞的積累和脂滴的增多增大,同時(shí)可以抑制由NF-κB炎癥信號(hào)通路引起的炎癥反應(yīng)。
[Abstract]:Perinatal period is the period of high incidence of energy metabolic disorders such as ketosis and fatty liver in dairy cows. The decrease of dry matter intake and the increase of energy demand in dairy cows lead to negative energy balance, which leads to fat mobilization and ketosis. Fatty liver and other disorders of energy metabolism. Therefore, fat mobilization caused by negative energy balance is an important part of energy metabolic disorders in dairy cows during perinatal period. In addition, adipose tissue is an important endocrine organ of the body. Many cytokines, such as tumor necrosis factor TNF- 偽, interleukin-1 尾 interleukin-1 尾 (IL-1 尾) and interleukin-6 (IL-6), are secreted. These cytokines are closely related to the development of energy metabolic disorders such as ketosis and fatty liver, as well as the occurrence and development of infectious diseases such as mastitis and endometritis. PLIN1 is a lipid coated protein that is widely expressed in adipocytes. It is important to understand the characteristics of lipid metabolism in adipose tissue of cow during perinatal negative energy balance, especially the changes in the expression and activity of lipid metabolism enzymes to alleviate fat mobilization. The key to prevent and cure perinatal disorders of energy metabolism is that PLIN1 may also play a special role in the expression and secretion of inflammatory factors in adipocytes. In order to reveal the mechanism of PLIN1 regulating lipid metabolism and inflammatory response of adipose tissue, the in vivo experiment and primary cultured adipocyte overexpression and silencing of PLIN1 were carried out in perinatal infected cattle in order to reveal the mechanism of regulation of PLIN1 on lipid metabolism in adipose tissue. In order to establish the theoretical foundation for the prevention and treatment of negative energy balance disease in the perinatal period, the primary adipocytes were isolated and cultured in vitro. The results showed that PLIN1 could significantly up-regulate the expression of SREBP-1c m RNA and protein and enhance the expression of key fatty acid synthase ACC-FAS-SCD, DGAT1 and DGAT2, and down-regulate the expression of PPAR 緯 m RNA and protein. Inhibiting the expression of HSL and ATGL resulted in the accumulation of lipid and the increase of lipid droplets in adipocytes. Silencing PLIN1 could significantly down-regulate the expression of SREBP-1c m RNA and protein and inhibit the expression of ACC-FAS-SCD, DGAT1 and DGAT2, and up-regulate the expression of PPAR 緯 m RNA and protein, and promote the expression of HSL and ATGL. In vitro, the primary adipocytes of dairy cattle were isolated and cultured, transfected with PLIN1 overexpression of adenovirus and silencing of si RNA, and then stimulated by LPS. Overexpression of PLIN1 could inhibit the activation of NF- 魏 B inflammatory signaling pathway, decrease the expression of inflammatory factors such as TNF 偽, IL-1 尾 and IL-6, and play an anti-inflammatory role, and after silencing PLIN1, the activation of NF- 魏 B inflammatory signaling pathway enhanced the expression of inflammatory factors such as TNF- 偽, IL-1 尾 and IL-6. The results showed that PLIN1 promoted the accumulation of TG in adipocytes and the increase of lipid droplets, and inhibited the inflammatory response induced by NF- 魏 B inflammatory signaling pathway.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S858.23

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本文編號(hào)：1647207