蛋氨酸对热处理下体外培养奶牛乳腺上皮细胞增殖、凋亡及相关基因表达的影响
发布时间：2018-07-23 16:42
【摘要】：本试验通过对不同浓度蛋氨酸作用于热处理后的体外培养奶牛乳腺上皮细胞进行研究,测定其对乳腺上皮细胞增殖、抗氧化性、凋亡及相关基因与蛋白表达方面的影响,从细胞水平探讨蛋氨酸对热应激下奶牛乳腺的保护作用。本试验分为两个部分:1、蛋氨酸对热处理及非热处理奶牛乳腺上皮细胞增殖、抗氧化性能的影响。用两组含不同浓度(0、30、60mg/L)蛋氨酸的DMEM培养基培养细胞2d后,37℃组用37℃水浴0.5h,42℃组(热处理组)42℃C水浴0.5h,恢复6h,通过MTT测定高温热处理对体外培养乳腺上皮细胞增殖的影响,培养液中丙二醛(MDA)、一氧化氮(NO)、一氧化氮合酶(NOS)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)的活性以及细胞凋亡率。结果表明,与37℃组相比,42℃组MDA含量、NO含量、NOS活力、LDH活性均升高;细胞活力、SOD活性、CAT活性下降,GSH-Px酶活力升高。与Omg/L蛋氨酸组相比,添加60mg/L蛋氨酸组的细胞活力升高(P0.05);LDH活性显著降低(P0.05);尤其 MDA、NO、NOS 含量极显著降低(P0.01);添加 30mg/L、60mg/L蛋氨酸能增加热处理下乳腺上皮细胞CAT活性(P0.05);SOD中,60mg/L蛋氨酸组含量最高(P0.05);培养液中GSH-Px活性,60mg/L蛋氨酸组最高(P0.01)。这说明42℃0.5h热处理使细胞产生了热应激,细胞受到氧化应激,有氧自由基含量上升。添加蛋氨酸可以有效缓解细胞热应激损伤,减少氧化应激。2、蛋氨酸对热处理及非热处理乳腺上皮细胞凋亡、相关基因及蛋白表达的影响。用含不同浓度(0、30、60mg/L)蛋氨酸的DMEM培养基培养细胞2d后,42℃C水浴0.5h后立即放回37℃恢复培养6h,测定细胞凋亡、HSF-1mRNA、HSP70mRNA、Bax/Bc1-2mRNA的表达、HSP70蛋白的表达以及Caspase-3的活性。结果表明,流式细胞仪测定细胞凋亡,与0mg/L蛋氨酸组相比,添加30、60mg/L蛋氨酸能显著降低细胞晚期凋亡/坏死率(P0.05),总死亡率差异极显著(P0.01);60mg/L蛋氨酸组HSF-1mRNA、HSP70mRNA表达量、HSP70蛋白的表达显著升高(P0.01);降低了 Bax/Bcl-2mRNA的表达量以及Caspase-3的活性(P0.01)。结论:培养基中添加60mg/L蛋氨酸能促进高温条件下乳腺上皮细胞增殖,起到抗凋亡的作用,提高HSF-1、HSP70的表达,降低Bax/Bcl-2表达量以及Caspase-3的活性,对细胞起到保护效应。
[Abstract]:In this study, the effects of methionine at different concentrations on the proliferation, antioxidation, apoptosis and the expression of related genes and proteins of bovine breast epithelial cells after heat treatment were studied, and the effects of methionine on the proliferation, antioxidation, apoptosis and the expression of related genes and proteins were measured. To investigate the protective effect of methionine on dairy cow mammary gland under heat stress at cell level. The experiment was divided into two parts: 1: 1. The effect of methionine on proliferation and antioxidation of mammary epithelial cells after heat treatment and non-heat treatment. The cells were cultured in DMEM medium containing different concentrations of methionine (0.30g / L) for 2 days. After 2 days, the cells were incubated in 37 鈩，
本文编号：2140010
[Abstract]:In this study, the effects of methionine at different concentrations on the proliferation, antioxidation, apoptosis and the expression of related genes and proteins of bovine breast epithelial cells after heat treatment were studied, and the effects of methionine on the proliferation, antioxidation, apoptosis and the expression of related genes and proteins were measured. To investigate the protective effect of methionine on dairy cow mammary gland under heat stress at cell level. The experiment was divided into two parts: 1: 1. The effect of methionine on proliferation and antioxidation of mammary epithelial cells after heat treatment and non-heat treatment. The cells were cultured in DMEM medium containing different concentrations of methionine (0.30g / L) for 2 days. After 2 days, the cells were incubated in 37 鈩，
本文编号：2140010
本文链接：https://www.wllwen.com/yixuelunwen/dongwuyixue/2140010.html
