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脂多糖对泌乳奶牛乳中氨基酸组成及蛋白质代谢相关基因表达的影响

DOI: 10.3969/j.issn.1006-267x.2012.09.021

Keywords: 脂多糖,乳腺上皮细胞,泌乳奶牛,氨基酸,蛋白质代谢

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Abstract:

本试验旨在通过半体内和体外试验研究灌注脂多糖(lipopolysacchride,LPS)对泌乳奶牛乳中氨基酸组成和蛋白质代谢相关基因表达的影响。半体内试验选用6头经产的荷斯坦奶牛,随机分为对照组和试验组,每组3头。采用交叉试验设计,每期正试期7d,间隔期14d;试验组阴外动脉灌注LPS(大肠杆菌型O111:B4,0.01μg/kg),对照组阴外动脉灌注生理盐水。体外试验以乳腺上皮细胞为模型,基础培养基中LPS添加水平分别为0、0.1和10.0ng/mL,培养24h后,采用实时定量-PCR(qRT-PCR)法检测目的基因的相对表达水平。结果表明,阴外动脉灌注LPS后,乳中必需氨基酸、支链氨基酸含量随着灌注时间的增加呈先升高后降低的趋势(P>0.05),灌注后6h达到最高;乳中非必需氨基酸含量随着灌注时间的增加呈先降低后升高的趋势(P>0.05),灌注后6h达到最低。LPS提高了乳腺上皮细胞中p70核糖体蛋白S6激酶-1(S6K1)和真核起始因子4E结合蛋白-1(4EBP1)mRNA的表达水平(P>0.05),且具有剂量效应。LPS对雷帕霉素靶点(mTOR)、Janus激酶2(JAK2)mRNA表达水平影响差异不显著(P>0.05),但有提高的趋势。与对照组相比,LPS显著提高了信号转导和转录激活因子5(STAT5)mRNA表达水平(P<0.05)。综上所述,LPS通过干扰mTOR通路和JAK2/STAT5通路影响了乳蛋白及氨基酸组成。

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