赖氨酸对奶牛乳腺上皮细胞内乳脂肪合成相关基因和蛋白表达的影响

发布时间：2018-04-29 06:24

本文选题：奶牛 + 乳腺上皮细胞　；参考：《动物营养学报》2017年09期

【摘要】：本试验旨在研究赖氨酸(Lys)对奶牛乳腺上皮细胞(BMECs)内乳脂肪合成相关基因和蛋白表达的影响,探讨Lys影响乳脂肪合成的机理。将第3代BMECs随机分为6组,每组6个重复,每个重复1个培养孔。各组培养基中Lys的浓度分别为0.5(基础培养基,对照)、1.0、2.0、4.0、8.0和16.0mmol/L,37℃、5%CO2培养48h后测定BMECs甘油三酯(TAG)含量、乳脂肪合成相关基因和蛋白的表达量。结果表明:BMECs内TAG含量(P=0.013)以及脂肪酸结合蛋白3(FABP3,P=0.001)、脂蛋白脂酶(LPL,P=0.096)、脂肪酸合成酶(FASN,P=0.003)、乙酰甘油磷酸脂酰转移酶6(AGPAT6,P=0.038)和甘油-3-磷酸酰基转移酶(GPAM,P=0.022)基因表达量对Lys呈显著或趋于显著的浓度依赖效应。FABP3基因表达量以2.0、4.0、8.0、16.0mmol/L组和LPL基因表达量以1.0、2.0、4.0、8.0、16.0 mmol/L组显著高于0.5mmol/L组(P0.05);FASN基因表达量以2.0mmol/L组最高,显著高于16.0mmol/L组(P0.05);硬脂酰辅酶A去饱和酶1(SCD1)基因表达量以2.0、4.0mmol/L组显著高于其他组(P0.05);磷脂酸磷酸酯酶1(LPIN1)、嗜乳脂蛋白亚家族1成员1(BTN1 A1)和黄嘌呤脱氢酶(XDH)基因表达量均以1.0、2.0、4.0、8.0mmol/L组显著高于0.5mmol/L组(P0.05);过氧化物酶体增殖物激活受体γ(PPARγ)基因及蛋白表达量均以2.0、4.0mmol/L组显著高于0.5和8.0、16.0mmol/L组(P0.05);固醇调节元件结合蛋白1(SREBP1)基因表达量以1.0、2.0、4.0mmol/L组显著高于其他组(P0.05),蛋白表达量以1.0 mmol/L组显著高于其他组(P0.05)。但高浓度Lys抑制AGPAT6和GPAM的基因表达,AGPAT6基因表达量以2.0、4.0、8.0、16.0mmol/L组显著低于0.5、1.0mmol/L组(P0.05),GPAM基因表达量以16.0mmol/L组显著低于0.5、1.0、2.0、4.0mmol/L组(P0.05)。可见,Lys对BMECs的乳脂肪合成具有显著的促进效果,但高浓度的Lys抑制了乳脂肪合成相关基因的表达。本试验条件下,培养基中Lys适宜浓度为2.0~4.0mmol/L。
[Abstract]:The purpose of this experiment was to investigate the effect of lysine (Lys) on the gene and protein expression of milk fat synthesis in dairy cow's mammary epithelial cells (BMECs), and to explore the mechanism of Lys affecting milk fat synthesis. Third generation of BMECs were randomly divided into 6 groups, 6 replicates in each group, and 1 culture holes per repeat. The concentration of Lys in each culture medium was 0.5 (basic medium, The content of BMECs triglyceride (TAG) and the expression of related genes and proteins of milk fat synthesis were measured by 1.0,2.0,4.0,8.0 and 16.0mmol/L, 37 C and 5%CO2 for 48h. The results showed that the content of TAG in BMECs (P=0.013), fatty acid binding protein 3 (FABP3, P=0.001), lipoprotein lipase, fatty acid synthetase, acetyl Gan The expression of oil phospholipid transferase 6 (AGPAT6, P=0.038) and glycerol -3- phosphate acyl transferase (GPAM, P=0.022) gene expressed a significant or significant concentration dependence effect on Lys,.FABP3 gene expression in 2.0,4.0,8.0,16.0mmol/L group and LPL gene expression in 1.0,2.0,4.0,8.0,16.0 mmol/L group was significantly higher than that of 0.5mmol/L group. The expression was highest in group 2.0mmol/L, significantly higher than that in group 16.0mmol/L (P0.05), and the expression of stearyl coenzyme A dehydrogenase 1 (SCD1) gene was significantly higher than that in other groups (P0.05); phosphatidic acid phosphatase 1 (LPIN1), 1 member of the fomiphin subfamily 1 (BTN1 A1) and xanthine dehydrogenase (XDH) gene expression were all 1.0,2.0,4.0,8.0mmol The /L group was significantly higher than the 0.5mmol/L group (P0.05), and the gene and protein expression of peroxisome proliferator activated receptor gamma (PPAR gamma) were significantly higher in 2.0,4.0mmol/L group than in group 0.5 and 8.0,16.0mmol/L (P0.05), and the expression of sterol regulatory element binding protein 1 (SREBP1) gene was significantly higher than other groups (P0.05) in 1.0,2.0,4.0mmol/L group (P0.05). The 1 mmol/L group was significantly higher than the other groups (P0.05), but the high concentration of Lys inhibited the gene expression of AGPAT6 and GPAM, and the expression of AGPAT6 gene in 2.0,4.0,8.0,16.0mmol/L group was significantly lower than that in 0.5,1.0mmol/L group (P0.05). The expression of GPAM gene in 16.0mmol/L group was significantly lower than that in 0.5,1.0,2.0,4.0mmol group. The high concentration of Lys inhibited the expression of genes related to milk fat synthesis. Under the experimental conditions, the suitable concentration of Lys in culture medium was 2.0~4.0mmol/L.

【作者单位】：内蒙古农业大学动物科学学院;
【基金】：国家奶业“973计划”项目(2011CB1008003)
【分类号】：S823.5

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