白细胞介素-1受体拮抗剂对脂多糖诱导的奶牛乳腺上皮细胞氧化损伤的保护作用  

Protective Effect of Interleukin⁃1 Receptor Antagonist on Lipopolysaccharide⁃Induced Oxidative Damage in Bovine Mammary Epithelial Cells

作　　者：郭咏梅[1] 石惠宇 闫素梅[1] 史彬林[1] 郭晓宇[1] 赵艳丽[1] GUO Yongmei;SHI Huiyu;YAN Sumei;SHI Binlin;GUO Xiaoyu;ZHAO Yanli(Key Laboratory of Animal Nutrition and Feed Science at Universities of Inner Mongolia Autonomous Region,College of Animal Science,Inner Mongolia Agricultural University,Hohhot 010018,China)

机构地区：[1]内蒙古农业大学动物科学学院,内蒙古自治区高校动物营养与饲料科学重点实验室,呼和浩特010018

出　　处：《动物营养学报》2023年第7期4596-4603,共8页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：内蒙古农业大学高层次人才科研启动项目(NDYB2020⁃4);国家自然科学基金项目(31160466);内蒙古农业大学基本科研业务费专项资金(BR220142)。

摘　　要：本试验以脂多糖(LPS)作为应激源,诱导奶牛乳腺上皮细胞(BMEC)氧化损伤,旨在探究白细胞介素-1受体拮抗剂(IL-1Ra)对LPS诱导的BMEC氧化损伤的保护作用,并确定抑制细胞氧化损伤的适宜IL-1Ra浓度。第3代贴壁生长的BMEC被随机分为12个组:对照组(无LPS,无IL-1Ra)、LPS组(1.0μg/mL LPS)、单独添加IL-1Ra(0.25、0.50、1.00、5.00和10.00 ng/mL)组、同时添加IL-1Ra与LPS(0.25 ng/mL IL-1Ra+1.0μg/mL LPS、0.50 ng/mL IL-1Ra+1.0μg/mL LPS、1.00 ng/mL IL-1Ra+1.0μg/mL LPS、5.00 ng/mL IL-1Ra+1.0μg/mL LPS、10.00 ng/mL IL-1Ra+1.0μg/mL LPS)组,每组设6个重复。不同处理的工作液作用于细胞6 h后,测定细胞相对增殖率(RGR),并且收集细胞和培养液测定抗氧化指标和炎性因子含量。结果显示:与对照组相比,单独添加不同浓度的IL-1Ra对RGR、谷胱甘肽过氧化物酶1(GPx1)、硫氧还蛋白还原酶1(TrxR1)、过氧化氢酶(CAT)活性与总抗氧化能力(T-AOC)以及丙二醛(MDA)、活性氧(ROS)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、诱导性一氧化氮合酶(iNOS)、一氧化氮(NO)含量均无显著影响(P>0.05);但是,单独添加不同浓度IL-1Ra对白细胞介素-1(IL-1)的生成有显著的抑制效果(P<0.05)。与LPS组相比,同时添加LPS和1.00 ng/mL IL-1Ra能显著提高RGR(P<0.05),显著增加GPx1和超氧化物歧化酶(SOD)活性与T-AOC,并显著降低IL-1、IL-6、TNF-α、MDA、ROS、iNOS、NO含量(P<0.05)。结果提示,单独添加IL-1Ra可以专一性的抑制BMEC内IL-1的产生,对细胞氧化应激的影响不显著;LPS加速了BMEC氧化还原水平的失衡,IL-1Ra可有效抑制LPS诱导的细胞氧化损伤,且IL-1Ra的适宜作用浓度为1.00 ng/mL。In this experiment,lipopolysaccharide(LPS)was used as the stimulus source to induce oxidative damage in bovine mammary epithelial cells(BMEC),to explore the protective effect of interleukin⁃1 receptor antagonist(IL⁃1Ra)on the oxidative damage in BMEC induced by LPS,and to determine the appropriate con⁃centration of IL⁃1Ra which inhibited the oxidative damage of cells.The third⁃generation BMEC was randomly divided into 12 groups using a single⁃factor complete randomized trial design,including the control group(without LPS and IL⁃1Ra),the LPS group(1.0μg/mL LPS),the only adding IL⁃1Ra(0.25,0.50,1.00,5.00 and 10.00 ng/mL)groups,and adding IL⁃1Ra+LPS(0.25 ng/mL IL⁃1Ra+LPS,0.50 ng/mL IL⁃1Ra+LPS,1.00 ng/mL IL⁃1Ra+LPS,5.00 ng/mL IL⁃1Ra+LPS,10.00 ng/mL IL⁃1Ra+LPS)groups,with 6 rep⁃licates per group.The cells were treated with different working fluids for 6 h,the relative growth rate(RGR)of cells was measured,and the cells and culture medium were collected to measure the antioxidant indices and inflammatory factor contents.The results showed that,compared with the control group,the RGR,the activi⁃ties of glutathione peroxidase 1(GPx1),thioredoxin reductase 1(TrxR1)and catalase(CAT),total antioxi⁃dant capacity(T⁃AOC),the contents of malondialdehyde(MDA),reactive oxygen species(ROS),tumor necrosis factor⁃α(TNF⁃α),interleukin⁃6(IL⁃6),induced nitric oxide(iNOS)and nitric oxide(NO)were not significantly influenced by treated with different concentrations of IL⁃1Ra alone(P>0.05);however,dif⁃ferent concentrations of IL⁃1Ra treated alone had a significant inhibitory effect on interleukin⁃1(IL⁃1)content(P<0.05).When contrasted to the LPS group,adding LPS and 1.00 ng/mL IL⁃1Ra significantly increased the RGR and the activities of GPx1 and superoxide dismutase(SOD)(P<0.05),and significantly decreased the contents of IL⁃1,IL⁃6,TNF⁃α,MDA,ROS,iNOS and NO(P<0.05).The results suggest that the addition of IL⁃1Ra alone specifically inhibits IL⁃1 production in BMEC and ha

关 键 词：奶牛乳腺上皮细胞 脂多糖 白细胞介素-1受体拮抗剂 

分 类 号：S811.3[农业科学—畜牧学]