| 摘要: |
| 为探索苜蓿黄酮对脂多糖（LPS）刺激下乳成分合成相关基因表达的影响，采用体外细胞培养技术，将奶牛乳腺上皮细胞分成4个组：1）基础培养基（Con）；2）基础培养基中加入1 μg/mL的LPS（L）；3）基础培养基中加入1 μg/mL 的LPS和75 μg/mL苜蓿黄酮（L+F）；4）基础培养基中加入75 μg/mL苜蓿黄酮（F）。细胞在37 ℃， 5% CO2的培养箱中培养12 h后，对其乳成分合成相关基因的表达进行测定。结果表明：1）相对于对照组，L组和L+F组的酪氨酸激酶（JAK2）表达显著降低（P<0.01），信号转导子和转录激活子（STAT5）和真核起始因子4E（eIF4E）表达显著升高（P<0.05）。L+F组真核细胞始动因子4E结合蛋白1（4EBP1）的表达显著高于F组（P<0.01），核糖体S6蛋白激酶1（S6K1）的表达显著高于L组 和对照组（P<0.05）。2）F组胆固醇调节元件结合蛋白1 （SREBP1）的表达显著低于L组（P<0.05），过氧化物酶体增殖物激活受体γ（PPAR-γ）的表达则相反；F组脂肪酸转运蛋白1（FATP1）和脂肪酸转运蛋白4（FATP4）的表达显著低于L和L+F组（P<0.01），脂肪酸结合蛋白3（FABP3）和乙酰辅酶A羧化酶（ACACA）的表达则相反；对照组硬脂酰辅酶A去饱和酶1（SCD1）的表达显著高于其他各组（P<0.01）。3） L+F组葡萄糖转运蛋白1（Glut1）（P<0.05）、葡萄糖转运蛋白4（Glut4）（P<0.05）和葡萄糖转运蛋白8（Glut8）（P<0.01）的表达显著高于对照组，F组己糖激酶2（HK2）的表达显著低于L组和L+F组（P<0.05），而β-1，4-半乳糖基转移酶（β-1，4-Gal T）的表达最低。因此，在无LPS刺激下，添加苜蓿黄酮可能会抑制乳脂合成；在LPS刺激下，添加苜蓿黄酮可能会促进乳蛋白和乳糖的合成。 |
| 关键词: 苜蓿黄酮 脂多糖 乳腺上皮细胞 奶牛 |
| DOI：10.11841/j.issn.1007-4333.2018.02.07 |
| 分类号: |
| 基金项目:国家自然科学基金项目（31572430）；江苏省高校优势学科建设工程项目（PAPD）；江苏省农业三新工程项目（SXGC[2016]326） |
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| Effects of alfalfa flavonoids on gene expression of milk component synthesis in bovine mammary epithelial cells induced by lipopolysaccharide |
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ZHAN Jinshun1,2, CHEN Xiaolian2, ZHAN Kang1, ZHAO Guoqi1
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1.College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;2.Institute of Animal Husbandry and Veterinary, Jiangxi Academy of Agricultural Sciences, Nanchang 330200, China
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| Abstract: |
| The aim of this study was to examine the effect of alfalfa flavonoids (AF) on the gene expression of milk component synthesis in bovine mammary epithelial cells (BMECs) induced by lipopolysaccharide(LPS).BMECs were cultured on media without LPS+AF (Con),and with 1 μ g/mL LPS (L),1 μ g/mL LPS+75 μ g/mL AF(L+F),75 μ g/mL AF(F) at 37℃ with 5% CO2 for 12 h.The expression of gene related to milk component synthesis was evaluated.The results showed:1) Compared with the control group,the L and L+F treatment groups showed significant decrease in JAK2 expression (P<0.01),but significant increase in the expressions of STAT5 and eIF4E (P<0.01).The relative expressions of 4EBP1 and S6K1 were significantly higher in the L+F treatment group than in the F treatment group (P<0.01),L treatment and control groups (P<0.05),respectively.2) The relative expression of SREBP1 was significantly lower in the F treatment group than that of the L treatment group (P<0.05),whereas the PPAR-γ showed the opposite result.The relative expressions of FATP1 and FATP4 were significantly lower in the F treatment group than in the L and L+F treatment groups (P<0.01),whereas the FABP3 and ACACA had the opposite result.The relative expression of SCD1 was significantly higher in the control group than in other groups (P<0.01).3) Compared with the control group,the L+F treatment group showed significantly higher expression levels of Glut1 (P<0.05), Glut8 (P<0.01) and Glut4 (P<0.05).The relative expression of HK2 was significantly lower in the F treatment group than in the L+F and L treatment groups (P<0.05),and the relative expression of β-1,4-Gal T was the lowest in the F treatment group.This study showed that alfalfa flavonoids might inhibit milk fat synthesis of BMECs cultured without LPS.Alfalfa flavonoids might promote lactose and protein synthesis of BMECs induced by LPS. |
| Key words: alfalfa flavonoids lipopolysaccharide mammary epithelial cells dairy cow |
