HT-2毒素对小鼠MII期卵母生殖细胞甲基化基因及DNA甲基化的影响  

Effects of HT-2 toxin on methylation genes and DNA methylation in germ cells of mice MII stage ocytes

作　　者：邓凯伟[1] 童翠红 DENG Kaiwei;TONG Cuihong(XinYang Agriculture and Forestry University,Xinyang,Henan Province 464000,China)

机构地区：[1]信阳农林学院,河南信阳464000

出　　处：《中国饲料》2020年第15期33-37,共5页China Feed

基　　金：信阳农林学院2017年度青年教师基金项目(201701005)。

摘　　要：为研究HT-2毒素对小鼠MII期卵母细胞甲基化基因及DNA甲基化的影响,试验将24只小鼠随机分为4组,根据饲料标准设置HT-2毒素浓度分别为1、1.5、2 mg/kg及空白对照组。连续灌胃16 d后进行超排,测定小鼠卵巢的重量,采集小鼠MII期卵母细胞,通过使用实时荧光定量PCR和激光共聚焦的方法对小鼠卵母细胞Tet1、Tet2、Tet3甲基化基因表达及DNA甲基化进行研究。结果表明:(1)2 mg/kg组小鼠卵巢重量较对照组极显著降低(P<0.01);2 mg/kg组较1、1.5 mg/kg组显著降低(P<0.05);其他各组间差异不显著(P>0.05)。(2)Tet1、Tet2基因表达水平试验组和对照组相比差异不显著(P>0.05);2 mg/kg组Tet3基因表达水平较对照组极显著降低(P<0.01);2 mg/kg组较1、1.5 mg/kg组显著降低(P<0.05);其他各组差异不显著(P>0.05)。(3)在激光共聚焦试验中,5-甲基胞嘧啶(5-mC)的荧光平均光密度(AO值)中,1.5、2 mg/kg组较对照组和1 mg/kg两组极显著提高(P<0.01);1.5 mg/kg组与2 mg/kg组差异极显著(P<0.01);其他组别间无显著差异(P>0.05)。5-羟甲基胞嘧啶(5-hmC)的AO值中,1.5、2 mg/kg组较对照组和1 mg/kg组极显著提高(P<0.01);其他各组差异不显著(P>0.05)。各梯度中5-mC和5-hmC平均光密度值间差异极显著(P<0.01)。综上所述,HT-2毒素可通过降低小鼠MII期卵母细胞Tet3基因的表达水平从而提高DNA甲基化水平。To study the effect of HT-2 toxin on methylation gene and DNA methylation in MII stage oocyte of mice,24 mice were randomly divided into four groups.According to the feed standard,the concentration of HT-2 toxin was 1,1.5,2 mg/kg and blank control group,respectively.After 16 days of continuous intragastric administration,the weight of mouse ovaries was measured.Mice MII oocytes were collected.The expression of Tet1,Tet2 and Tet3 methylation genes and DNA methylation in mouse oocytes were studied by real-time fluorescence quantitative PCR and laser confocal method.The results showed that:(1)The ovarian weight of mice in 2 mg/kg group was significantly lower than that of control group(P<0.01);2 mg/kg group was significantly lower than that in 1 mg/kg and 1.5 mg/kg groups(P<0.05);there was no significant difference among other groups(P>0.05).(2)There was no significant difference in Tet1 and Tet2 gene expression level between the experimental group and the control group(P>0.05);the Tet3 gene expression level in 2 mg/kg group was significantly lower than that in the control group(P<0.01);the 2 mg/kg group was significantly lower than that in 1 mg/kg and 1.5 mg/kg groups(P<0.05);the other groups had no significant difference(P>0.05).(3)In laser confocal test,the average fluorescence density(AO value)of 5-methoxycytosine(5-mC)in 1.5,2 mg/kg groups was significantly higher than in the control group and 1 mg/kg group(P<0.01);the 1.5 mg/kg group was significantly higher than that in 2 mg/kg group(P<0.01);there was no significant difference between the other groups(P>0.05).Among the AO values of 5-hydroxymethoxycytosine(5-hmC),the 1.5,2 mg/kg groups was significantly higher than that in control group and 1 mg/kg group(P<0.01),but there was no significant difference among other groups(P>0.05).The average optical densities of 5-mC and 5-hmC were significantly different among the gradients(P<0.01).In conclusion that HT-2 toxin could improve DNA methylation by reducing Tet3 gene expression in MII mouse oocyte.

关 键 词：HT-2毒素 卵母细胞 DNA甲基化 生殖性能 

分 类 号：S816.3[农业科学—饲料科学]