本文選題：維生素A　切入點(diǎn)：乙酸　出處：《內(nèi)蒙古農(nóng)業(yè)大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：本試驗旨在研究維生素A(V_A)對奶牛乳腺上皮細(xì)胞(BMECs)內(nèi)乳脂乳蛋白合成相關(guān)基因表達(dá)的影響,并在此基礎(chǔ)上添加乙酸,進(jìn)而觀察在補(bǔ)充了乳脂肪合成前體物的條件下,二者對乳成分合成的影響是否存在互作關(guān)系,以期能深入了解V_A對乳脂乳蛋白合成的影響機(jī)制,為奶牛生產(chǎn)中V_A的合理添加以及改善乳脂乳蛋白的合成提供依據(jù)。試驗一采用單因子完全隨機(jī)試驗設(shè)計,將貼壁生長的第三代BMECs以2×10~5個/mL的密度接種于6孔板中繼續(xù)培養(yǎng),待其貼壁率達(dá)到80%左右時隨機(jī)分為6個處理組,每組6個重復(fù)。使用無血清培養(yǎng)基饑餓24 h后,添加V_A濃度分別為0、0.05、0.1、0.2、1.0 和 2.0μg/mL 的細(xì)胞培養(yǎng)液,0μg/mLV_A 為對照組,繼續(xù)培養(yǎng)24h后收集細(xì)胞,進(jìn)行相關(guān)指標(biāo)的測定。試驗結(jié)果表明:].0、2.0μg/mL V_A可以顯著提高BMECs細(xì)胞活力以及甘油三酯(TG)的合成;V_A能上調(diào)乳脂肪合成調(diào)控因子過氧化酶體增殖物激活受體(PPARG)、固醇調(diào)節(jié)元件結(jié)合蛋白1(SREBP1)、硬脂酰輔酶A去飽和酶(SCD)的基因表達(dá);V_A也能促進(jìn)信號轉(zhuǎn)導(dǎo)轉(zhuǎn)錄激活因子5(STAT5)、αsl-酪蛋白(CSN1S1)、K-酪蛋白(CSN3)等乳蛋白合成相關(guān)基因的相對表達(dá)。試驗二采用二因素完全隨機(jī)試驗設(shè)計,將貼壁生長的第三代BMECs隨機(jī)分為6個處理組,每組6個重復(fù)。使用無血清培養(yǎng)基饑餓處理24 h后,分別添加不同V_A濃度和乙酸濃度(乙酸鈉為原料)的工作液,6種工作液的濃度分別是:0μg/mLV_A+0 mmol/L 乙酸,0μg/mL V_A+6 mmol/L 乙酸,0 μg/mL V_A+10 mmol/L乙酸,1 μg/mLV_A+Ommol/L 乙酸,1 μg/mL V_A+6 mmol/L 乙酸,1 μg/mLV_A+10 mmol/L乙酸;0μg/mLV_A+O mmol/L乙酸為對照組。繼續(xù)培養(yǎng)24 h后按照測定指標(biāo)要求的方法進(jìn)行細(xì)胞收集和相關(guān)指標(biāo)的測定。試驗結(jié)果表明:V_A和乙酸在乳脂合成方面存在互作效應(yīng),乙酸能提高BMECs內(nèi)的TG含量,但同時添加V_A則抑制TG合成;乙酸能上調(diào)BMECs內(nèi)乙酰輔酶A羧化酶(ACACA)、脂肪酸合成酶(FASN)的活性及PPARG基因的表達(dá),同時添加V_A則起抑制作用;乙酸能顯著提高BMECs內(nèi)哺乳動物雷帕霉素靶蛋白(mTOR)含量、S6K1酶活性,以及上調(diào)CSN3基因的表達(dá),V_A與乙酸在mTOR與CSN2的基因表達(dá)方面具有互作效應(yīng),二者同時添加會減弱單一添加V_A或乙酸時對mTOR與CSN2的基因表達(dá)的下調(diào)作用。綜合考慮,0V_A+10乙酸組對乳脂合成的促進(jìn)作用較好,1V_A+10乙酸組較弱;OV_A+6乙酸組對乳蛋白合成促進(jìn)作用較好,1V_A+6乙酸組較弱。
[Abstract]:The purpose of this study was to investigate the effect of vitamin A / V _ (V _ (A)) on the expression of genes related to milk fat milk protein synthesis in milk epithelial cells (BMECs) of dairy cows, and to add acetic acid on the basis of this, and then observe the condition of supplementation of precursor of milk fat synthesis. Whether there is an interaction between the two effects on the synthesis of milk ingredients, in order to understand the mechanism of the effect of VSAT on the synthesis of milk milk protein, In experiment 1, the third generation BMECs of adherent growth was inoculated in 6 hole plate with density of 2 脳 10 ~ 5 / mL, and the third generation of BMECs was inoculated in 6 hole plate with the density of 2 脳 10 ~ 5 / mL. When the adherence rate was about 80%, each group was randomly divided into six treatment groups, each group was subjected to 6 replicas. After starvation in serum-free medium for 24 hours, the cells were collected after 24 hours of starvation with 0 0. 05 渭 g / mL and 2. 0 渭 g / mL of Va, respectively. The results showed that:] 0 渭 g / mL Vapa could significantly increase the viability of BMECs cells and the synthesis of triglyceride (TG) could up-regulate the expression of lipid synthesis regulator, peroxisome proliferator activator receptor (PPARGN) and steroid regulator. The gene expression of stearyl coenzyme A desaturase (SCD1) could also promote the relative expression of signal transduction activator 5 (STAT5), 偽 sl-casein 1 (CSN1S1) K- casein casein (CSN3) and other genes related to milk protein synthesis. Experiment 2 used two factors to study the expression of genes related to milk protein synthesis, such as protein binding protein 1 (SREBP1) and stearyl coenzyme A desaturase (SCD). Complete random trial design, The third generation BMECs of adherent growth was randomly divided into 6 treatment groups with 6 replicates in each group. The concentration of six kinds of working liquid with different VA concentration and different concentration of acetic acid (sodium acetate as raw material) were 1: 0 渭 g / mLVA0 mmol/L, 0 渭 g / mLVA6 mmol/L acetic acid = 0 渭 g / mLVA 10 mmol/L acetic acid, 1 渭 g / mLVA Ommol/L acetic acid 1 渭 g / mLVA Ommol/L Ommol/L 1 渭 g / mLVA / 6 mmol/L / mmol/L, 1 渭 g / mLVA = 10 渭 g / mLVA = 10 渭 g / mLVA = 10 渭 g / mLVA = 1 渭 g / mLVA = 1 渭 g / mLVA = 1 渭 g / mLVA10. Mmol/L acetic acid 0 渭 g / mLVA O mmol/L acetic acid was used as the control group. After 24 hours of culture, cell collection and related indexes were determined according to the method required for the determination of the indexes. The results showed that there was an interaction effect between the two groups in the synthesis of milk fat. Acetic acid could increase TG content in BMECs, but at the same time add VA to inhibit TG synthesis, acetic acid could upregulate the activity of acetyl coA carboxylase, fatty acid synthase (FASN) and the expression of PPARG gene in BMECs. Acetic acid could significantly increase the activity of S6K1 enzyme in mammalian rapamycin target protein mTORand up-regulate the expression of CSN3 gene in BMECs. VA and acetic acid could interact with each other in the gene expression of mTOR and CSN2. The down-regulation of gene expression of mTOR and CSN2 was weakened by the addition of both of them. Considering the effect of VA10 acetic acid on the synthesis of milk fat, the effect of VA10 acetic acid on milk fat synthesis was better. 1 VA10 acetic acid group was weaker than OVA6 acetic acid group on milk protein. The effect of synthesis was better than that of 1 V A 6 acetic acid group.
【學(xué)位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S823

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