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香烟烟雾对小鼠卵母细胞染色质构型、DNA甲基化及相关基因表达的影响 被引量:1

Effects of cigarette smoke on chromatin configuration,DNA methylation and expression of related genes in mice oocytes
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摘要 目的探讨香烟烟雾暴露对小鼠卵母细胞染色质构型、全基因组DNA甲基化及相关基因表达的影响。方法将清洁级健康ICR雌鼠分成3组(对照组和香烟烟雾低剂量组、高剂量组),分别置于点燃0、1、2支香烟的染毒柜(容积18 L)中,每天2次,每次1 h,共4周。利用Hoechst 33342对细胞核染色,观察卵母细胞质量及染色质构型;间接免疫荧光法分析卵母细胞全基因组DNA甲基化模式,实时荧光定量PCR测定卵母细胞DNA甲基转移酶(DNA methyltransferases,DNMT)1、DNMT3a及DNMT3b基因的mRNA表达水平。结果随着烟雾浓度的升高,去透明带卵母细胞直径显著减小(P<0.05),核仁未包围(non surrounded nucleolus,NSN)染色质构型比例明显升高(P<0.05)。高剂量组卵母细胞DNA甲基化水平显著下降(P<0.05)。随烟雾浓度升高DNMT1表达量降低,DNMT3b在高剂量组中表达水平显著降低(P<0.05)。结论香烟烟雾暴露可能通过改变卵母细胞染色质构型,降低DNA甲基转移酶的表达,使DNA甲基化水平下降,从而降低卵母细胞质量。 Objective To study the effect of cigarette smoke on chromatin configuration,DNA methylation and expression of related genes in mice oocytes. Methods The clean grade ICR female mice were divided into 3 groups,control group,low dose group and high dose group. Animals were placed inside a perspex chamber( 18L) filled up with the smoke produced by 0,1,2 cigarettes,one hour a time,twice daily for 4weeks. Nuclear staining with Hoechst 33342,to observe the quality of oocyte and chromatin configuration. Analysis of the DNA methylation patterns by indirect immunofluorescence. The expressions of DNA methyltransferases( DNMT) 1,DNMT3a and DNMT3b mRNA were detected by quantitative real time PCR. Results With the increase of smoke concentration,the diameter of oocytes was significantly decreased( P〈0. 05),the percentage of non surrounded nucleolus( NSN) chromatin configuration was significantly increased( P〈0. 05). In high dose group,the level of DNA methylation wassignificantly lower than that of control group( P〈0. 05). The expression of DNMT1 decreased with the increase of smoke concentration,but the expression level of DNMT3b was significantly decreased( P〈0. 05). Conclusion Cigarette smoke exposure may alter oocyte chromatin configuration,decrease the expression level of DNMTs,resulted in oocytes DNA methylation decreased,so as to decrease the quality of oocytes.
出处 《卫生研究》 CAS CSCD 北大核心 2017年第2期213-218,共6页 Journal of Hygiene Research
基金 安徽省教育厅自然科学研究重点项目(No.KJ2016A735) 皖南医学院校中青年自然科学研究项目(No.WK201511)
关键词 香烟烟雾 染色质构型 DNA甲基化 小鼠卵母细胞 cigarette smoke chromatin configuration DNA methylation mouse oocytes
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