过氧化氢诱导的奶牛乳腺上皮细胞氧化损伤模型的建立
首发时间:2014-06-20
摘要:本试验旨在利用过氧化氢(H2O2)作为刺激源,以细胞存活率及抗氧化指标为判断指标,研究建立奶牛乳腺上皮细胞(BMEC)的氧化损伤模型。(方法)试验分两部分进行。试验一采用单因子完全随机试验设计,将第三代贴壁生长的BMEC随机分为42个处理组,每个处理组10个重复。细胞中分别添加0、100、200、400、600、800和1 000 μmol/L H2O2,使之分别作用2、4、6、8、12、24 h,以测定其对细胞存活率的影响。试验二是在试验一得出的适宜H2O2处理时间(本试验的结果为6h)的基础上,采用单因子随机试验设计,将第三代贴壁生长的BMEC随机分为7个处理组,每个处理组6个重复,BMEC中添加不同浓度的H2O2(0、100、200、400、600、800和1 000 μmol/L)作用细胞6 h,收集细胞和培养液测定抗氧化指标,筛选使得细胞发生氧化损伤的适宜H2O2处理浓度。(结果)结果表明,600 μmol/L H2O2作用细胞6 h,BMEC的存活率降低至79.79%;600~1 000μmol/L H2O2处理组与对照组相比均显著降低了谷胱甘肽过氧化物酶、超氧化物歧化酶和过氧化氢酶活性,提高了丙二醛含量(P<0.05),与400μmol/L H2O2处理组比较差异显著(P<0.05),但600、800和1 000 μmol/L H2O2处理组之间差异不显著(P>0.10)。(结论)结果提示,H2O2作用浓度为600 μmol/L、作用时间为6 h,对BMEC产生了明显的氧化应激,可作为建立细胞氧化损伤模型时的适宜剂量与作用时间。
关键词: 动物营养学 奶牛乳腺上皮细胞 过氧化氢 氧化损伤模型
For information in English, please click here
Establishment of Oxidative Damage Model of Bovine Mammary Epithelial Cells Induced by Hydrogen Peroxide
Abstract:This study was conducted to determine the effect of hydrogen peroxide (H2O2) on cell viability and antioxidant paramaters to establish the oxidative damage model of bovine mammary epithelial cells (BMEC). The experiment was divided into two parts. The experiment 1 was conducted as a single factor randomized arrangement, and the 3th passage BMEC were divided into 42 groups with 10 replicates in each group. Cells were incubated in different concentrations of H2O2 (0, 100, 200, 400, 600, 800 and 1 000 μmol/L) for 2 h, 4 h, 6 h, 8 h, 12 h and 24 h in order to explore the effect of concentrations and action time of H2O2 on the cell viability. On the basis of the experiment 1, the experiment 2 was also conducted as a single factor randomized arrangement, and the 3th passage BMEC were divided into 7 groups with 6 replicates in each group. Culture mediums containing different concentrations of H2O2 (0, 100, 200, 400, 600, 800 and 1 000 μmol/L) for 6 h were used in each treatment, then cells and the medium were collected for the analysis of antioxidant paramaters, to select the appropriate concentration of H2O2 ensuring oxidative damage to cells. The results showed as follows: the BMEC survival reduced to 79.79% when the cells were induced by 600 μmol/L H2O2 for 6 h. Cell induced by 600-1 000 μmol/L H2O2 significantly decreased the activities of glutathione peroxidase, superoxide dismutase and catalase, significantly increased malondialdehyde content compared with the control group or 400 μmol/L H2O2 treatment group(P<0.05). But the difference between 600, 800 and 1 000 μmol/L H2O2 treatment groups was not significant (P> 0.10). When H2O2 concentration is increased to 600 μmol/L and its reaction time lasts 6 h, a significant oxidative stress is produced in BMEC, which is used to establish a standard model of oxidative stress.
Keywords: animal nutrition bovine mammary epithelial cells hydrogen peroxide oxidative damage model
论文图表:
引用
No.4601092921287140****
同行评议
勘误表
过氧化氢诱导的奶牛乳腺上皮细胞氧化损伤模型的建立
评论
全部评论0/1000