摘要
试验旨在探究在H_(2)O_(2)诱导的氧化应激状态下,超氧化物歧化酶模拟物(SODm)对仔猪空肠上皮细胞系(IPEC-J2)的保护作用。利用MTT法筛选出构建氧化应激模型H_(2)O_(2)的适宜浓度;将IPEC-J2细胞分别用0、0.05、0.5、5、50、500、2000 U/mL SODm进行培养,利用MTT法分别在2、4、8、12 h时测定各组细胞存活率,筛选出SODm作用的适宜浓度和时间;根据构建的氧化应激模型和SODm适宜浓度和时间,将IPEC-J2细胞随机分为空白组、模型组、SODm处理组(SODm_(0.5)、SODm_(5)、SODm_(50))和超氧化物歧化酶(SOD)处理组(SOD_(0.5)、SOD_(5)、SOD_(50)),分别测定各组细胞存活率、活性氧(ROS)含量及细胞内SOD、谷胱甘肽过氧化物酶(GSH-Px)活性及丙二醛(MDA)含量和总抗氧化能力(T-AOC)。结果表明:①H_(2)O_(2)构建细胞氧化应激模型的适宜浓度是1.0 mmol/mL。②当不同浓度SODm分别作用4、8、12 h时,0.5~50 U/mL SODm组细胞存活率显著高于空白组(P<0.05);且8 h时存活率最高,在12 h时处于下降趋势。③除空白组外,SODm5和SOD5预处理的IPEC-J2存活率显著高于其他组(P<0.05);且SODm和SOD组中细胞ROS含量显著低于模型组(P<0.05);模型组与空白组相比,均显著降低了SOD、GSH-Px活性和T-AOC水平,提高了MDA含量(P<0.05);与模型组相比,所有SODm和SOD处理组均显著提高了SOD、GSH-Px活性和T-AOC水平,降低了MDA含量(P<0.05),同时SODm_(50)组T-AOC水平显著低于SOD50组(P<0.05)。综上,SODm对H_(2)O_(2)诱导氧化损伤状态下IPEC-J2具有保护作用。
The objective of this study was to investigate the protective effect of superoxide dismutase mimetic(SODm)on intestinal epithelial cell(IPEC-J2)under H_(2)O_(2)-induced oxidative stress.Using MTT method to screen out the proper concentration of H_(2)O_(2)applied in structuring oxidative stress model.IPEC-J2 were cultured with 0,0.05,0.5,5,50,500 and 2000 U/mL SODm,respectively.The survival of each group was detected by MTT in 2,4,8 and 12 h to determine the proper range and duration of a SODm dose.Based on the model of oxidative stress and the proper range and duration of SODm,IPEC-J2 were randomly grouped as the control group,the model group with H_(2)O_(2),the disposed group with SODm(SODm_(0.5),SODm_(5),SODm_(50)),the disposed group with SOD(SOD_(0.5),SOD_(5),SOD_(50)),then cell viability,ROS levels and SOD,GSH-Px,MDA,T-AOC levels in cells were measured in each group.The results were showed as follows:①1.0 mmol/mL H_(2)O_(2)was the proper concentration in structuring oxidative stress model.②The survival ratios of the disposed group with SODm of 0.5~50 U/mL in 4,8 and 12 h was significantly higher than that of the control groups(P<0.05).The survival ratio was highest at 8 h and decreased at 12 h.③The IPEC-J2 survival rate of the SODm5 and SOD5 groups were significantly higher than that in other groups(P<0.05)except for the blank group,and the ROS contents in the SODm and SOD groups were significantly lower than that in the model group(P<0.05).The activities of SOD,GSH-Px and T-AOC were significantly decreased,and MDA content were significantly increased in model group compared with the control group(P<0.05).The treatments of SODm and SOD increased the activities of SOD,GSH-Px and T-AOC,and significantly decreased MDA content compared with the model group(P<0.05),The T-AOC level of SOD50 group was higher than that of SODm_(50)group(P<0.05).In conclusion,SODm had protective effect on IPEC-J2 cells under H_(2)O_(2)-induced oxidative stress.
作者
王思博
崔红霞
杨季
许丽
WANG Sibo;CUI Hongxia;YANG Ji;XU Li(College of Animal Science and Technology, Northeast Agricultural University,Harbin 150030,China;Inner Mongolia Ordos City Animal Husbandry Station,Ordos 017000,China)
出处
《中国畜牧兽医》
CAS
北大核心
2021年第5期1566-1573,共8页
China Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金项目(32072749)。
