本文選題：肉仔雞 + 糖皮質(zhì)激素　； 參考：《山東農(nóng)業(yè)大學》2017年碩士論文

【摘要】：FATP1作為脂肪酸轉(zhuǎn)運蛋白,在機體脂肪酸轉(zhuǎn)運攝取方面發(fā)揮著重要作用。本論文從"脂肪酸轉(zhuǎn)運影響脂肪沉積"的角度出發(fā),利用活體和離體試驗,在核酸、蛋白、細胞、組織等水平系統(tǒng)研究了FATP1在糖皮質(zhì)激素(DEX)調(diào)控肉雞骨骼肌脂肪異位沉積中的作用及機制。1.FATP1在應(yīng)激肉雞骨骼肌脂肪代謝中的作用。處理組根據(jù)肉雞體重注射DEX。結(jié)果表明,DEX處理會導(dǎo)致肉雞肌肉組織FATP1的mRNA水平升高,脂肪沉積量增多,血液中VLDL和INS含量顯著升高。腿肌糖皮質(zhì)激素受體(GR)、脂聯(lián)素受體(ADPNR)、過氧化物酶體增殖物激活受體α(PPARα)的mRNA水平也升高。由這些結(jié)果推測,DEX調(diào)控骨骼肌的FATP1促進脂肪攝取可能是通過GR-ADPNR-PPARα通路實現(xiàn)的。2.脂肪水平對糖皮質(zhì)激素調(diào)控FATP1的影響。分別用不同脂肪水平的日糧飼喂肉雞,35日齡時用DEX處理。試驗結(jié)果表明,低脂日糧飼喂肉雞組用DEX處理后肌肉組織TG沉積量增多,但高脂日糧飼喂肉雞組用DEX處理后各組織脂肪沉積量無顯著變化;低脂日糧飼喂肉雞并用DEX處理后,肉雞肌肉組織GR、ADPNR、PPARα和FATP1的mRNA水平與組織TG沉積量有相同變化趨勢。但是,這種趨勢在飼喂高脂日糧并用DEX處理后的肉雞腿肌上表現(xiàn)不明顯。由此可以推出,不同日糧處理下DEX對肌肉GR-ADPNR-PPARα-FAPT1通路基因水平的影響并不一致。3.脂肪酸類型對糖皮質(zhì)激素調(diào)控FATP1的影響。利用添加了相同濃度油酸和棕櫚酸的條件培養(yǎng)基分別培養(yǎng)成肌細胞,細胞分化至80%左右分別用不同濃度的DEX處理。結(jié)果表明,用飽和脂肪酸培養(yǎng)的細胞用DEX處理后,細胞脂肪沉積增多,而不飽和脂肪酸培養(yǎng)的細胞用DEX處理后細胞脂肪沉積量有所下降;檢測細胞AKT蛋白的磷酸化水平,結(jié)果顯示DEX處理對其無顯著影響;油酸培養(yǎng)細胞DEX處理后檢測到相關(guān)基因的mRNA水平無顯著差異,而棕櫚酸培養(yǎng)的成肌細胞用DEX處理后檢測到GR、ADPNR、PPARα和FATP1的mRNA水平變化趨勢與細胞中TG沉積量的變化趨勢基本一致。這說明不同脂肪酸類型會影響DEX對FATP1的調(diào)控作用,進而影響細胞TG的沉積量。4.糖皮質(zhì)激素通過FATP1調(diào)控成肌細胞脂肪沉積的途徑。單獨DEX處理成肌細胞,發(fā)現(xiàn)FATP1和PPARα的mRNA水平無顯著變化;利用添加了相同濃度油酸和棕櫚酸的條件培養(yǎng)基分別培養(yǎng)成肌細胞,細胞分化至80%左右分別利用特異性抑制劑RU486、U0126、GW6471特異性抑制GR、ERK2/MAPK和PPARα在細胞中的作用,并用DEX處理。發(fā)現(xiàn)油酸培養(yǎng)的成肌細胞用抑制劑和DEX處理后脂肪酸攝取無顯著差異,棕櫚酸培養(yǎng)的成肌細胞用抑制劑處理后脂肪酸攝取能力下降。這說明DEX會通過GR-PPARα途徑或ERK2/MAPK-PPARα途徑調(diào)控細胞對飽和脂肪酸的攝取。綜上所述,糖皮質(zhì)激素可能通過升高FATP1的mRNA水平促進細胞對脂肪酸的轉(zhuǎn)運攝取,進而導(dǎo)致脂肪沉積。日糧不同脂肪水平會影響到糖皮質(zhì)激素通過FATP1對肉雞骨骼肌脂肪沉積的調(diào)控作用,細胞培養(yǎng)底物的脂肪酸類型也會影響到糖皮質(zhì)激素通過FATP1對成肌細胞脂肪沉積的調(diào)控作用。糖皮質(zhì)激素應(yīng)激條件下,細胞傾向于通過GR-PPARα或ERK2/MAPK-PPARα通路攝取飽和脂肪酸。本研究揭示了應(yīng)激導(dǎo)致骨骼肌脂肪異位沉積的調(diào)控靶點,闡明了日糧脂肪與糖皮質(zhì)激素的相互作用。
[Abstract]:FATP1, as a fatty acid transporter, plays an important role in the transport of fatty acid in the body. From the point of view of "fatty acid transport affecting adipose deposition", this paper studies the regulation of FATP1 by Glucocorticoid (DEX) in the skeletal muscle of Broilers by living and in vitro experiments at the level of nucleic acid, protein, cell and tissue. The role of.1.FATP1 in the lipid metabolism in the skeletal muscle of broilers. The results of the treatment group by injection of DEX. according to the body weight of broiler showed that DEX treatment could lead to the increase of mRNA level in the muscle tissue of the broiler, the increase of the fat deposition, the increase of the content of VLDL and INS in the blood. The leg muscle glucocorticoid receptor (GR) and the adiponectin were affected. The mRNA level of the peroxisome proliferator activated receptor alpha (PPAR alpha) is also elevated in the body (ADPNR). From these results, it is speculated that the DEX regulation of the FATP1 of skeletal muscle may be the effect of the.2. fat level on the glucocorticoid regulated FATP1 through the GR-ADPNR-PPAR alpha pathway. The diets of broilers were fed on the diet of different fat levels, 35 DEX treatment was used at the age of age. The results showed that the amount of TG deposition in the muscle tissue was increased after DEX treatment in the low fat diet diet, but there was no significant change in the fat deposition in the tissues of the broiler group fed with DEX. The mRNA levels of GR, ADPNR, PPAR A and FATP1 in broiler muscle tissues were compared with those of low fat diet fed broilers and DEX treated with DEX. TG deposition in tissue has the same trend of change. However, this trend is not obvious on the meat chicken leg muscles treated with high fat diet and DEX treatment. Therefore, the effect of DEX on the GR-ADPNR-PPAR alpha -FAPT1 pathway gene level in different diets does not agree with the effect of.3. fatty acid type on the regulation of FATP1 in glucocorticoids. The myocytes were cultured with the condition medium added with the same concentration of oleic acid and palmitic acid. The cell differentiation to about 80% was treated with different concentrations of DEX. The results showed that after the cells cultured with saturated fatty acids were treated with DEX, the cell fat deposition was increased, and the cells cultured in unsaturated fatty acid were treated with DEX after DEX treatment. The deposition of AKT protein was decreased, and the results showed that the DEX treatment had no significant effect on it. The mRNA level of the related genes was not significantly different after DEX treatment in oleic acid culture cells, and the mRNA level of GR, ADPNR, PPAR A and FATP1 in the myoblast cultured by palmitic acid was detected by DEX. The change trend of TG deposition is basically the same. This shows that different types of fatty acids can affect the regulation of DEX on FATP1, and then influence the deposition of TG,.4. glucocorticoid, through FATP1 regulation of myoblast fat deposition. The single DEX treatment of myoblasts, found that FATP1 and PPAR alpha mRNA levels have no significant changes; use of addition to the use of addition. The condition medium with the same concentration of oleic acid and palmitic acid were cultured into myocytes, and the cells differentiated to about 80% by specific inhibitors RU486, U0126, GW6471, the role of GR, ERK2/MAPK and PPAR alpha in the cells, and DEX treatment. The fatty acid uptake of the myoblasts cultured by oleic acid and after DEX treatment was found. There is no significant difference between the palmitic acid cultured myoblasts and the decrease in fatty acid uptake by inhibitors. This suggests that DEX regulates the uptake of saturated fatty acids through the GR-PPAR alpha pathway or ERK2/MAPK-PPAR alpha pathway. To sum up, glucocorticoids may promote the uptake of fatty acids by increasing the mRNA level of FATP1. And then lead to fat deposition. Different levels of dietary fat may affect the regulation of glucocorticoid through FATP1 on the fat deposition of skeletal muscle in broilers. The type of fatty acids in cell culture substrates also affects the regulation of glucocorticoid through FATP1 on the adipose deposition of myoblasts. Under glucocorticoid stress conditions, cell tendencies This study reveals the regulatory target of stress induced adipose deposition in skeletal muscle and elucidates the interaction between dietary fat and glucocorticoid through the uptake of saturated fatty acids through the GR-PPAR alpha or ERK2/MAPK-PPAR alpha pathway.

【學位授予單位】：山東農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S831

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