褪黑素对H_(2)O_(2)诱导蛋鸡卵巢颗粒细胞氧化应激损伤的保护作用  

Protective Effect of Melatonin on Oxidative Stress Injury of Ovarian Granulosa Cells Induced by H_(2)O_(2) in Laying Hens

作　　者：白康[1] 郝二英 任河陵 艾嘉伟 宁炜 黄仁录[1] 陈辉[1] BAI Kang;HAO Erying;REN Heing;AI Jiawei;NING Wei;HUANG Renu;CHEN Hui(College of Animal Science and Technology,Hebei Agricultural University,Baoding,Hebei 071000;Chengdu Little Giant Animal Husbandry Equipment Co.,Ltd.,Chengdu,Sichuan 611435)

机构地区：[1]河北农业大学动物科技学院,河北保定071000 [2]成都小巨人畜牧设备有限公司,四川成都611435

出　　处：《中国家禽》2024年第9期1-7,共7页China Poultry

基　　金：河北省自然科学基金项目(C2022204229);现代农业产业技术体系建设专项资金(CARS-40);保定市科技计划项目(2211N014)。

摘　　要：为探讨褪黑素对蛋鸡卵巢颗粒细胞氧化应激损伤的保护作用,试验选取产蛋高峰期蛋鸡进行颗粒细胞分离培养,分别用0、12.5、25、50、100、200µmol/L H_(2)O_(2)处理3、6、9 h,根据颗粒细胞增殖活性和氧化指标变化建立氧化应激模型。试验分为4组:试验Ⅰ组为对照组,试验Ⅱ组为H_(2)O_(2)组,试验Ⅲ组为H_(2)O_(2)+Mel组,试验Ⅳ组为褪黑素组,利用CCK-8法、DCF法及TUNEL法分别检测各组细胞增殖率、活性氧(ROS)水平和凋亡率,q-PCR和Western blot检测抗氧化指标SOD1、SOD2、CAT、GPX3和PRDX3基因和蛋白的表达水平。结果显示:①与对照组相比,H_(2)O_(2)组细胞增殖率降低(P<0.05),丙二醛(MDA)水平增加(P<0.05),过氧化氢酶(CAT)、超氧化物歧化酶(SOD)活性下降(P<0.05);②与对照组相比,H_(2)O_(2)组细胞活性氧(ROS)水平、凋亡率增加(P<0.05),增殖活性降低(P<0.05),Ⅲ组细胞活性氧水平、凋亡率降低(P<0.05),增殖活性增加(P<0.05);③与对照组相比,H_(2)O_(2)组细胞SOD1、SOD2、CAT、GPX3和PRDX3等抗氧化基因和蛋白表达下降(P<0.05),而褪黑素干预后则促进了抗氧化基因和蛋白表达(P<0.05)。研究提示:褪黑素干预后缓解了H_(2)O_(2)对颗粒细胞的氧化应激损伤,其可通过清除活性氧及上调蛋鸡卵巢颗粒细胞抗氧化基因和蛋白的表达起保护作用。In order to investigate the protective effect of melatonin on ovarian granulosa cells damaged by oxidative stress,granulosa cells isolated from laying hens during the peak laying period were treated with 0,12.5,25,50,100 and 200µmol/L H_(2)O_(2) for 3,6 and 9 h respectively.The oxidative stress model of granulosa cells was established according to the cell proliferation activity and the changes of oxidative indexes.The experiment was divided into 4 groups:test groupⅠwas the control group;test groupⅡwas H_(2)O_(2) group;Test groupⅢwas H_(2)O_(2)+Mel group;Test groupⅣwas the melatonin group.The CCK-8,TUNEL and DCF method were used to detect cell proliferation,apoptosis,and active oxygen(ROS)levels in each group,respectively,while q-PCR and Western blot were used to detect the expression levels of antioxidant indicators SOD1,SOD2,CAT,GPX3 and PRDX3 genes and proteins.The results showed as follows:①Compared with test groupⅠ,the cell proliferation rate,activities of catalase(CAT)and superoxide dismutase(SOD)decreased in H_(2)O_(2) group(P<0.05),the level of malondialdehyde(MDA)was increased(P<0.05);②Compared with test groupⅠ,the ROS level and apoptosis rate of cells increased in H_(2)O_(2) group(P<0.05)and the pro-liferative activity was decreased(P<0.05),while the ROS level and apoptosis rate of cells in test groupⅢwere decreased(P<0.05)and the proliferative activity was increased(P<0.05).③Compared with the test groupⅠ,the expression of antioxidant genes and proteins such as SOD1,SOD2,CAT,GPX3 and PRDX3 in H_(2)O_(2) group were decreased(P<0.05),while the expression of antioxi-dant genes and proteins was promoted after melatonin intervention(P<0.05).The results indicated that melatonin could alleviate the oxidative stress damage of H_(2)O_(2) on granulosa cells,and it could play a protective role by clearing ROS and up regulating the expression of antioxidant genes and proteins in granulosa cells of layers'ovaries.

关 键 词：褪黑素 蛋鸡 卵巢颗粒细胞 氧化应激 保护 

分 类 号：S831.2[农业科学—畜牧学]