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苜蓿黄酮对脂多糖诱导下奶牛乳腺上皮细胞凋亡的影响
引用本文:占今舜,陈小连,詹康,苏效双,赵国琦.苜蓿黄酮对脂多糖诱导下奶牛乳腺上皮细胞凋亡的影响[J].草业学报,2018,27(1):187-194.
作者姓名:占今舜  陈小连  詹康  苏效双  赵国琦
作者单位:扬州大学动物科学与技术学院,江苏扬州225009;江西省农业科学院畜牧兽医研究所,江西南昌330200;江西省农业科学院畜牧兽医研究所,江西南昌,330200;扬州大学动物科学与技术学院,江苏扬州,225009
基金项目:国家自然科学基金项目,现代农业产业技术体系专项资金,江苏省高校优势学科建设工程项目
摘    要:研究苜蓿黄酮对脂多糖(LPS)诱导下奶牛乳腺上皮细胞凋亡的影响。将奶牛乳腺上皮细胞分成4个组,即基础培养基、基础培养基中加入1 μg·mL-1的LPS、基础培养基中加入1 μg·mL-1的LPS和75 μg·mL-1苜蓿黄酮、基础培养基中加入75 μg·mL-1苜蓿黄酮。细胞在37 ℃, 5% CO2的培养箱中培养。结果表明:1)LPS刺激12 h后奶牛乳腺上皮细胞活性下降,而添加苜蓿黄酮能够极显著抑制LPS诱导下细胞活性的下降(P<0.01)。2)在LPS刺激下,细胞内的活性氧(ROS)浓度升高,而添加苜蓿黄酮能够显著降低其浓度(P<0.05)。3)LPS显著上调细胞的IL-1β、IL-6、TNF-α、TLR2、TLR4和MyD88表达(P<0.01),而苜蓿黄酮能够显著下调细胞的IL-1β、IL-6、TNF-α和TLR2表达(P<0.01或P<0.05)。4)在LPS刺激下,p53、Caspase3、p38和P-p38蛋白的表达显著升高(P<0.01或P<0.05),而添加苜蓿黄酮能够显著降低p53和p38蛋白的表达(P<0.05)。在LPS诱导下,苜蓿黄酮能够通过降低ROS浓度,抑制细胞凋亡,提高细胞活性;可能通过抑制TLR2/MyD88信号通路来降低细胞炎症因子的表达,从而保护细胞免受炎性损伤。

关 键 词:苜蓿黄酮  脂多糖  奶牛乳腺上皮细胞  细胞凋亡
收稿时间:2017-03-07
修稿时间:2017-04-10

Effects of alfalfa flavonoids on apoptosis of bovine mammary epithelial cells induced by lipopolysaccharide
ZHAN Jin-shun,CHEN Xiao-lian,ZHAN Kang,SU Xiao-shuang,ZHAO Guo-qi.Effects of alfalfa flavonoids on apoptosis of bovine mammary epithelial cells induced by lipopolysaccharide[J].Acta Prataculturae Sinica,2018,27(1):187-194.
Authors:ZHAN Jin-shun  CHEN Xiao-lian  ZHAN Kang  SU Xiao-shuang  ZHAO Guo-qi
Institution: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
Abstract:The aim of this study was to examine the effect of alfalfa flavonoids (AF) on apoptosis of bovine mammary epithelial cells (BMECs) induced by lipopolysaccharide (LPS).The BMECs were exposed to 4 treatments;medium containing 0 μg · mL 1 LPS and AF (control),1 μg · mL-1 LPS (L),1 μg · mL-1 LPS and 75 μg · mL-1 AF(L+F),and 75 μg · mL-1 AF(F),respectively.The BMECs were cultured in cell incubator at 37 ℃,5% CO2.The results were as follow:1) AF supplementation significantly reduced the viability of BMECs stimulated by LPS for 12 h (P<0.01).2) LPS significantly increased the concentration of ROS in cells (P<0.01),whereas AF supplementation significantly reduced the concentration of ROS in cells induced by LPS (P<0.05).3) LPS significantly increased the relative expression of IL-1β,IL-6,TNF-α,TLR2,TLR4 and MyD88 in cells (P<0.01),but the relative expression of IL-1β,IL-6,TNF-a and TLR2 in cells induced by LPS was significantly reduce by AF supplementation (P<0.01 or P<0.05).4) LPS significantly increased the expression of p53,Caspase3,p38 and P-p38 proteins in cells (P<0.01 or P<0.05),whereas AF supplementation inhibited the expression of p53 and p38 proteins in cells induced by LPS (P<0.05).The results showed that AF could improve the viability of cells and inhibit apoptosis by reducing the concentration of ROS and that AF might play a role in protecting cells against inflammatory injure by inhibiting the TLR2/MyD88 signaling pathway.
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