苜蓿黄酮对脂多糖诱导下奶牛乳腺上皮细胞凋亡的影响

doi:10.11686/cyxb2017098

草业学报 ›› 2018, Vol. 27 ›› Issue (1): 187-194.DOI: 10.11686/cyxb2017098

苜蓿黄酮对脂多糖诱导下奶牛乳腺上皮细胞凋亡的影响

占今舜^{1, 2}, 陈小连², 詹康¹, 苏效双¹, 赵国琦^{1, *}

1.扬州大学动物科学与技术学院,江苏扬州 225009;
2.江西省农业科学院畜牧兽医研究所,江西南昌 330200

收稿日期:2017-03-07 修回日期:2017-04-10 出版日期:2018-01-20 发布日期:2018-01-20
通讯作者: E-mail:gqzhao@yzu.edu.cn
作者简介:占今舜(1985-),男,江西玉山人,博士。E-mail:zhanjinshun1985@163.com
基金资助:
国家自然科学基金项目(No.31572430),现代农业产业技术体系专项资金(CARS-36)和江苏省高校优势学科建设工程项目(PAPD)资助

Effects of alfalfa flavonoids on apoptosis of bovine mammary epithelial cells induced by lipopolysaccharide

ZHAN Jin-shun^{1, 2}, CHEN Xiao-lian², ZHAN Kang¹, SU Xiao-shuang¹, ZHAO Guo-qi^{1, *}

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

Received:2017-03-07 Revised:2017-04-10 Online:2018-01-20 Published:2018-01-20

摘要/Abstract

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。

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-α 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.

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.

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.

参考文献

[1] Piotrowska-Tomala K K, Siemieniuch M J, Szóstek A Z, et al . Lipopolysaccharides, cytokines, and nitric oxide affect secretion of prostaglandins and leukotrienes by bovine mammary gland epithelial cells. Domestic Animal Endocrinology, 2012, 43(4): 278-288.
[2] Shi H, Guo Y, Liu Y, et al . The in vitro effect of lipopolysaccharide on proliferation, inflammatory factors and antioxidant enzyme activity in bovine mammary epithelial cells. Animal Nutrition, 2016, 2(2): 99-104.
[3] Sun Y, Li L, Wu J, et al . Bovine recombinant lipopolysaccharide binding protein (BRLBP) regulated apoptosis and inflammation response in lipopolysaccharide-challenged bovine mammary epithelial cells (BMEC). Molecular Immunology, 2015, 65(2): 205-214.
[4] Zhu L P, Yang W H, Ren T T, et al . The expression of NF-κB and IκB-α induced by LPS in bovine mammary epithelial cells. Chinese Journal of Veterinary Medicine, 2016, 52(5): 14-17.
朱丽萍, 杨文浩, 任婷婷, 等. 脂多糖诱导奶牛乳腺上皮细胞中炎性因子和核转录子- κB的表达. 中国兽医杂志, 2016, 52(5): 14-17.
[5] Testai L. Flavonoids and mitochondrial pharmacology: A new paradigm for cardioprotection. Life Sciences, 2015, 135: 68-76.
[6] Zhang C, Song Y, Zhang Y, et al . Antagonistic action to inhibition of lipopolysaccharide-treated chondrocyte proliferation and anti-inflammatory action of genistein. Chinese Journal of Biologicals, 2011, 24(2): 184-186.
张城, 宋勇, 张艺, 等. 金雀异黄酮对脂多糖处理的软骨细胞增殖抑制的拮抗及抗炎作用. 中国生物制品学杂志, 2011, 24(2): 184-186.
[7] Jiang N, Zhou X B, Ou G W, et al . Effects of 4', 5, 7 trihydroxyisoflavone on mRNA expression of ephrinB2 and IL-6 induced by LPS in human aortic en-dothelial cells. Journal of Modern Medicine & Health, 2016, 32(7): 966-968.
姜念, 周雪冰, 欧刚卫, 等. 4', 5, 7-三羟基异黄酮对LPS诱导的人主动脉内皮细胞ephrinB2及IL-6 mRNA表达的影响. 现代医药卫生, 2016, 32(7): 966-968.
[8] Zhang X, Guo Z, Zhang C, et al . Effect of total flavonoids of Astragalus on secretion levels of cytokines in LPS-induced RAW264.7 cell. Chinese Veterinary Science, 2015, 45(3): 321-324.
张贤, 郭泽, 张冲, 等. 黄芪总黄酮对脂多糖体外诱导的RAW264.7细胞的细胞因子和NO分泌水平的影响. 中国兽医科学, 2015, 45(3): 321-324.
[9] Gao W W. Study on the Bioactivity of Alfalfa and the Influence Factors to Its Flavonoids and Saponins. Beijing: Chinese Peking Union Medical College, 2004.
高微微. 苜蓿生物活性及影响其黄酮和皂苷成分因素的研究. 北京: 中国协和医科大学, 2004.
[10] Su X S, Zhan J S, Zhan K, et al . Proliferation stimulus and antioxidant effect of alfalfa flavonoids on dairy cow mammary epithelial cells cultured in vitro . Acta Prataculturae Sinica, 2015, 24(12): 139-145.
苏效双, 占今舜, 詹康, 等. 苜蓿黄酮对体外培养的奶牛乳腺上皮细胞增殖与抗氧化的影响. 草业学报, 2015, 24(12): 139-145.
[11] Zhan J S, Wei M J, Su X S, et al . Effects of alfalfa flavonoids on bovine mammary epithelial cell cultures in vitro under heat stress. Acta Prataculturae Sinica, 2016, 25(4): 159-165.
占今舜, 魏明吉, 苏效双, 等. 苜蓿黄酮对热应激下体外培养奶牛乳腺上皮细胞凋亡的影响. 草业学报, 2016, 25(4): 159-165.
[12] Zhan J S, Liu M M, Zhan K, et al . Effects of alfalfa flavonoids on lactoprotein, milk fat and lactose synthesis in bovine mammary epithelial cells. Chinese Journal of Animal Science, 2017, 53(1): 91-95.
占今舜, 刘明美, 詹康, 等. 苜蓿黄酮对奶牛乳腺上皮细胞乳蛋白、乳脂和乳糖合成的影响. 中国畜牧杂志, 2017, 53(1): 91-95.
[13] Zhan J S, Liu M M, Su X S, et al . Effects of alfalfa flavonoids on the production performance, immune system, and ruminal fermentation of dairy cows. Asian-Australasian Journal of Animal Sciences, 2017, 30(10): 1416-1424.
[14] Liu L X, Lin Y, Zhang L, et al . Cytotoxicity of LPS and effects on milk protein synthesis in dairy cow mammary epithelial cells. Journal of Northeast Agricultural University, 2015, 46(6): 61-66.
刘立新, 林叶, 张莉, 等. 脂多糖对奶牛乳腺上皮细胞毒性作用及乳蛋白合成的影响. 东北农业大学学报, 2015, 46(6): 61-66.
[15] Wang H, Meng X, Qiu C W, et al . The innate immune responses of bovine mammary epithelial cells stimulated by lipopolysaccharide. Chinese Journal of Veterinary Science, 2010, 30(3): 398-401.
王亨, 孟霞, 邱昌伟, 等. 脂多糖诱导奶牛乳腺上皮细胞先天性免疫反应. 中国兽医学报, 2010, 30(3): 398-401.
[16] Chen J, Wang G L, Li L, et al . Effects of estrogen on proliferation and antioxidation in bovine ( Bos taurus ) mammary epithelial cells. Journal of Agricultural Biotechnology, 2013, 21(2): 216-222.
陈静, 王根林, 李莲, 等. 雌激素对奶牛乳腺上皮细胞增殖及抗氧化性的影响. 农业生物技术学报, 2013, 21(2): 216-222.
[17] Zhu J M, Li N, Zhang Y J, et al . The research progress of alfalfa flavonoids. Prataculturae Science, 2009, 26(9): 156-162.
朱见明, 李娜, 张亚军, 等. 苜蓿黄酮的研究进展. 草业科学, 2009, 26(9): 156-162.
[18] Chanput W, Krueyos N, Ritthiruangdej P. Anti-oxidative assays as markers for anti-inflammatory activity of flavonoids. International Immunopharmacology, 2016, 40: 170-175.
[19] Wang X Q, Ji G Y, Jiang Z Q. Effects of genistein on inflammatory factors, adenosine monophosphate-activated protein kinase phosphorylation in lipopolysaccharides-treated RAW264. 7 cells. Acta Nutrimenta Sinica, 2012, 34(2): 177-180.
王小倩, 纪桂元, 蒋卓勤. 染料木黄酮对脂多糖诱导的RAW264. 7细胞炎症因子、腺苷酸激活蛋白激酶磷酸化的影响. 营养学报, 2012, 34(2): 177-180.
[20] Zhang D D, Ling S, Zhang H P, et al . The study on mechanism of the total flavonoids from Daphne genkwa . Shanghai Journal of Traditional Chinese Medicine, 2010, 8: 58-62.
章丹丹, 凌霜, 张洪平, 等. 芫花总黄酮的抗炎机制研究. 上海中医药杂志, 2010, 8: 58-62.
[21] Gao M, Ao Y, Luan X H. Progress on negative regulatory mechanism of Toll-like receptor signal transduction. Progress in Veterinary Medicine, 2015, 36(1): 96-101.
高明, 敖越, 栾新红. Toll样受体信号转导的负调控机制研究进展. 动物医学进展, 2015, 36(1): 96-101.
[22] Li H, Wang Q, Chen X M, et al . Mangiferin inhibits lipopolysaccharide-induced production of interleukin-6 in human oral epithelial cells by suppressing toll-like receptor signaling. Archives of Oral Biology, 2016, 71: 155-161.
[23] Feng G, Sun B, Li T Z. Daidzein attenuates lipopolysaccharide-induced acute lung injury via toll-like receptor 4/NF-kappaB pathway. International Immunopharmacology, 2015, 26(2): 392-400.
[24] Sun J Y, Li D L, Yan D, et al . Baicalin inhibits toll-like receptor 2/4 expression and downstream signaling in rat experimental periodontitis. International Immunopharmacology, 2016, 36: 86-93.
[25] Chen H, Mo X, Yu J, et al . Alpinetin attenuates inflammatory responses by interfering toll-like receptor 4/nuclear factor kappa B signaling pathway in lipopolysaccharide-induced mastitis in mice. International Immunopharmacology, 2013, 17(1): 26-32.
[26] Zhang C H, Wang Y H, Wang C W, et al . Reactive oxygen species participated in the activation induced cell apoptosis of mice macrophage RAW264.7 provoked by LPS. Chinese Journal of Veterinary Medicine, 2014, 50(11): 24-27.
张宸豪, 王月华, 王长文, 等. 活性氧参与脂多糖诱导小鼠巨噬细胞RAW264.7活化诱导凋亡. 中国兽医杂志, 2014, 50(11): 24-27.
[27] Deng W, Liu Y, Chen C G, et al . Effect of hesperetin on the level of reactive oxygen speries in H9C2 cardiomyocytes which induced by lipopolysaccharide. Journal of Medical Research, 2014, 43(11): 36-38.
邓伟, 刘源, 陈昌贵, 等. 橙皮素对脂多糖诱导H9C2心肌细胞活性氧水平的影响. 医学研究杂志, 2014, 43(11): 36-38.
[28] Li W J, Pan Q J, Li M Y. Research progress of p53 gene and its function. Letters in Biotechnology, 2014, 25(2): 282-285.
李文娟, 潘庆杰, 李美玉. p53基因及其功能研究进展. 生物技术通讯, 2014, 25(2): 282-285.
[29] Zhuang Q Y, Liu J, Han J H. Functions and mechanisms of the p38 MAP Kinase pathway. Chinese Journal of Cell Biology, 2013, 35(2): 123-133.
庄秋宇, 刘俊, 韩家淮. p38丝裂原活化蛋白激酶的功能与调控机制. 中国细胞生物学学报, 2013, 35(2): 123-133.

苜蓿黄酮对脂多糖诱导下奶牛乳腺上皮细胞凋亡的影响

Effects of alfalfa flavonoids on apoptosis of bovine mammary epithelial cells induced by lipopolysaccharide

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