分子营养 Molecular Nutrition

苜蓿素对脂多糖刺激下体外培养奶牛乳腺上皮细胞活性和炎症相关基因表达的影响

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  • 江西省农业科学院畜牧兽医研究所, 南昌 330200
占今舜(1985-),男,江西玉山人,助理研究员,博士,主要从事反刍动物微生态营养及分子营养。E-mail:zhanjinshun1985@163.com

收稿日期: 2017-08-21

  网络出版日期: 2018-02-02

基金资助

草畜一体化关键技术研究与示范(JXXTCX201702-04)

Effects of Tricin on Viability and Expressions of Genes Related to Inflammation of Bovine Mammary Epithelial Cells Stimulated by Lipopolysaccharide in Vitro

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  • Institute of Animal Husbandry and Veterinary, Jiangxi Academy of Agricultural Sciences, Nanchang 330200, China

Received date: 2017-08-21

  Online published: 2018-02-02

摘要

本试验旨在研究苜蓿素对脂多糖(LPS)刺激下体外培养奶牛乳腺上皮细胞活性和炎症相关基因表达的影响。将体外培养的奶牛乳腺上皮细胞分成6组,对照组采用基础培养基,试验组在基础培养基中分别加入1 μg/mL LPS(L组)、1 μg/mL LPS和2.5 μg/mL苜蓿素(L+2.5组)、1 μg/mL LPS和5.0 μg/mL苜蓿素(L+5组)、1 μg/mL LPS和10.0 μg/mL苜蓿素(L+10组)以及1 μg/mL LPS和15.0 μg/mL苜蓿素(L+15组),培养24 h。结果表明:1)与对照组相比,L组奶牛乳腺上皮细胞相对增殖率极显著降低(P<0.01),而L+2.5组和L+10组则极显著升高(P<0.01);2)与L组相比,L+10组和L+15组细胞超氧化物歧化酶活性显著升高(P<0.05),而乳酸脱氢酶活性和一氧化氮浓度显著降低(P<0.05);3)与L组相比,L+5组和L+10组细胞的白细胞介素8、白细胞介素1β、肿瘤坏死因子α、Toll样受体2和Toll样受体4的相对表达量均极显著降低(P<0.01),L+5组细胞的髓样分化因子88相对表达量均显著降低(P<0.05)。结果提示,在LPS刺激下,添加苜蓿素能够提高奶牛乳腺上皮细胞活性和抗氧化性能,抑制炎症相关基因的表达,发挥抗炎作用。

本文引用格式

占今舜, 谷德平, 胡利珍, 钟小军, 霍俊宏 . 苜蓿素对脂多糖刺激下体外培养奶牛乳腺上皮细胞活性和炎症相关基因表达的影响[J]. 动物营养学报, 2018 , 30(2) : 641 -648 . DOI: 10.3969/j.issn.1006-267x.2018.02.029

Abstract

The aim of this experiment was to investigate the effects of tricin on viability and expressions of genes related to inflammation of bovine mammary epithelial cells (BMECs) stimulated by lipopolysaccharide (LPS) in vitro. The bovine mammary epithelial cells cultured in vitro were divided into 6 groups and cultured for 24 h. The BMECs cultured in basal culture medium were used as the control group. The BMECs cultured in basal culture medium supplemented with 1 μg/mL LPS, 1 μg/mL LPS+2.5 μg/mL tricin, 1 μg/mL LPS+5.0 μg/mL tricin, 1 μg/mL LPS+10.0 μg/mL tricin and 1 μg/mL LPS+15.0 μg/mL tricin were used as L, L+2.5, L+5, L+10 and L+15 groups, respectively. The results showed as follow:1) compared with the control group, the relative growth rate of BMECs in L group was significantly decreased (P<0.01), whereas that in L+2.5 and L+10 groups was significantly increased (P<0.01); 2) compared with L group, the activity of superoxide dismutase of cells was significantly increased (P<0.05), but the activity of lactate dehydrogenase and the concentrations of nitric oxide and malondialdehyde of cells were significantly reduced in L+10 and L+15 groups (P<0.05); 3) compared with L group, the relative expression levels of interleukin-8, interleukin-1β, tumor necrosis factor-α, Toll-like receptor 2 and Toll-like receptor 4 in L+5 and L+10 groups were significantly decreased (P<0.01), and the relative expression level of myeloid differentiation primary response 88 in L+5 group was significantly decreased (P<0.05). The results indicated that the supplementation of tricin can enhance the viability and oxidation resistance of BMECs stimulated by LPS, and plays a role of anti-inflammatory effect by inhibiting gene expressions related to inflammation.

参考文献

[1] MORITA-TAKEMURA S,NAKAHARA K,TATSUMI K,et al.Changes in endothelial cell proliferation and vascular permeability after systemic lipopolysaccharide administration in the subfornical organ[J].Journal of Neuroimmunology,2016,298:132-137.

[2] 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)[J].Molecular Immunology,2015,65(2):205-214.  

[3] SHI H Y,GUO Y M,LIU Y,et al.The in vitro effect of lipopolysaccharide on proliferation,inflammatory factors and antioxidant enzyme activity in bovine mammary epithelial cells[J].Animal Nutrition,2016,2(2):99-104.  

[4] 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[J].中国兽医杂志,2016,52(5):14-17,49.

[5] ZHOU J M,IBRAHIM R K.Tricin-a potential multifunctional nutraceutical[J].Phytochemistry Reviews,2010,9(3):413-424.  

[6] CHUNG I M,HAHN S J,AHMAD A.Confirmation of potential herbicidal agents in hulls of rice,Oryza sativa[J].Journal of Chemical Ecology,2005,31(6):1339-1352.  

[7] YAZAWA K,KUROKAWA M,OBUCHI M,et al.Anti-influenza virus activity of tricin,4',5,7-trihydroxy-3',5'-dimethoxyflavone[J].Antiviral Chemistry & Chemotherapy,2011,22(1):1-11.  

[8] BICHOFF E M,LIVINGSTON A L,BOOTH A N.Tricin from alfalfa:isolation and physiological activity[J].Journal of Pharmaceutical Sciences,1964,53(11):1411-1412.  

[9] 陈艳,姚宏,林新华.苜蓿素对人直肠癌SW1116细胞增殖和凋亡的作用及其机制[J].天然产物研究与开发,2013,25(1):31-35.

[10] 刘利艳,殷玉婷,陈力,等.苜蓿素对小鼠免疫功能的影响[J].广东医学,2016,37(8):1118-1121.

[11] LI F Y,WANG W,CAO Y G,et al.Inhibitory effects of astragalin on lipopolysaccharide-induced inflammatory response in mouse mammary epithelial cells[J].Journal of Surgical Research,2014,192(2):573-581.  

[12] WANG J J,GUO C M,WEI Z K,et al.Morin suppresses inflammatory cytokine expression by downregulation of nuclear factor-κB and mitogen-activated protein kinase (MAPK) signaling pathways in lipopolysaccharide-stimulated primary bovine mammary epithelial cells[J].Journal of Dairy Science,2016,99(4):3016-3022.  

[13] 任婷婷,张东君,朱丽萍,等.葛根素对奶牛乳腺上皮细胞炎症模型中NF-κB信号通路的影响[J].农业生物技术学报,2016,24(1):44-51.

[14] 詹康,贡笑笑,左晓昕,等.奶牛乳腺上皮细胞系的培养与鉴定[J].动物营养学报,2015,27(8):2544-2550.

[15] 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[J].草业学报,2016,25(4):159-165.

[16] 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[J].中国畜牧杂志,2017,53(1):91-95.

[17] 刘春龙,李忠秋,张帆,等.大豆黄酮和染料木素对体外培养奶牛乳腺上皮细胞增殖及抗氧化水平的影响[J].畜牧兽医学报,2008,39(11):1517-1522.

[18] 穆莹,江连洲.大豆异黄酮对奶牛乳腺上皮细胞增殖及泌乳性能的影响[J].中国畜牧兽医,2013,40(1):187-190.

[19] 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[J].草业学报,2015,24(12):139-145.

[20] 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[J].东北农业大学学报,2015,46(6):61-66.

[21] 刘立新,张羽男,张磊,等.紫花苜蓿总黄酮对LPS诱导乳腺上皮细胞炎性因子的抑制作用[J].黑龙江医药科学,2016,39(4):1-3.

[22] 肖定福,钟佳,刘进辉,等.壳寡糖对脂多糖诱导猪空肠上皮细胞氧化损伤的作用[J].动物营养学报,2016,28(7):2090-2095.

[23] NGUYEN D H,ZHAO B T,LE D D,et al.Phenolic constituents and their anti-inflammatory activity from Echinochloa utilis grains[J].Natural Product Sciences,2016,22(2):140-145.  

[24] LIU X C,ZHENG L,Li Q F,et al.Naringin protects against lipopolysaccharide-induced cardiac injury in mice[J].Environmental Toxicology and Pharmacology,2016,48:1-6.

[25] 刘翠翠,赵龙,石晓岚,等.茶多酚对脂多糖诱导的支气管上皮细胞损伤的保护作用及其机制[J].中国儿童保健杂志,2017,25(10):1015-1018.

[26] 汪娟,蒋维,王毅.降香中黄酮类化合物对脂多糖诱导的RAW264.7细胞抗炎作用研究[J].细胞与分子免疫学杂志,2013,29(7):681-684.

[27] KANG B M,AN B K,JUNG W S,et al.Anti-inflammatory effect of tricin isolated from Alopecurus aequalis Sobol. on the LPS-induced inflammatory response in RAW 264.7 cells[J].International Journal of Molecular Medicine,2016,38(5):1614-1620.  

[28] 王秀梅,姚菲菲,孙立秋,等.麦冬对脂多糖作用下大鼠腹膜间皮细胞TNF-α和TGF-β1表达分泌的影响[J].中医药学报,2012,40(2):17-20.

[29] 王海波,刘玉兰,李权,等.脂多糖对仔猪下丘脑-垂体-肾上腺轴内应激基因和NOD信号通路关键基因表达的影响[J].中国畜牧杂志,2015,51(1):20-24.

[30] 涂治骁,刘玉兰,吴欢听,等.脂多糖刺激对仔猪肠道、肝脏和肌肉组织NODs信号通路关键基因及其负调控因子mRNA表达的影响[J].中国畜牧杂志,2016,52(11):35-38,54.

[31] SHALINI V,BHASKAR S,KUMAR K S,et al.Molecular mechanisms of anti-inflammatory action of the flavonoid,tricin from Njavara rice (Oryza sativa L.)in human peripheral blood mononuclear cells:possible role in the inflammatory signaling[J].International Immunopharmacology,2012,14(1):32-38.  

[32] SHALINI V,JAYALEKSHMY A,HELEN A.Mechanism of anti-inflammatory effect of tricin,a flavonoid isolated from Njavara rice bran in LPS induced hPBMCs and carrageenan induced rats[J].Molecular Immunology,2015,66(2):229-239.  

[33] SHALINI V,PUSHPAN C K,SINDHU G,et al.Tricin,flavonoid from Njavara reduces inflammatory responses in hPBMCs by modulating the p38MAPK and PI3K/Akt pathways and prevents inflammation associated endothelial dysfunction in HUVECs[J].Immunobiology,2016,221(2):137-144.  

[34] 殷玉婷,刘丽丽,刘利艳.苜蓿素对哮喘小鼠肺泡巨噬细胞TLR4/MyD88/NF-κB通路的抑制作用[J].中成药,2017,39(3):450-454.
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