CdCl_2对牛卵母细胞体外成熟及凋亡的影响  

The Influence of Bovine Oocytes in Vitro Maturation and Apoptosis by CdCl_2

作　　者：刘涛[1] 李向臣[1] 关伟军[1] 马月辉[1] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所,北京100193

出　　处：《华北农学报》2011年第5期146-152,共7页Acta Agriculturae Boreali-Sinica

基　　金："863"高新技术发展计划(2006AA10Z198);转基因生物新品种培育科技重大专项(2008ZX08009-003);国家科技支撑项目(2006BAD13B08;2008BADB2B01)

摘　　要：利用不同浓度(0,0.8,1.6,2.4μg/mL)的氯化镉(CdCl2)对处理牛卵母细胞前后对其成熟率、凋亡率、Ca2+分布、线粒体膜电位、Bcl-2与Bax的基因表达的影响,初步为重金属对雌性哺乳动物生殖细胞发育及细胞凋亡机制的进一步研究打下基础。利用0,0.8,1.6,2.4μg/mL的氯化镉(CdCl2)处理牛卵母细胞22 h,统计其凋亡率及死亡率;利用Annexin V-FITC检测牛卵母细胞的早期凋亡;分别用Fluo-3/AM和Rh123检测卵母细胞的Ca2+分布及其线粒体膜电位的变化;利用RT-PCR检测促凋亡基因Bcl-2和抑凋亡基因Bax的相对表达。经过上述处理后,0.8μg/mL处理组卵母细胞的成熟率和凋亡率与对照组相比差异不显著(P>0.05),而1.6μg/mL和2.4μg/mL处理组与对照组相比差异显著(P<0.05);随着处理浓度的增加,胞浆Ca2+的荧光强度逐渐增强,线粒体膜电位逐渐下降,bcl-2与Bax的相对比值下降。CdCl2诱导卵母细胞的凋亡呈剂量效应,可以抑制卵母细胞的成熟,促进细胞的凋亡。In this research, bovine oocytes were dealed with different concentration of CdC12 （0,0.8,1.6,2.4 g/mL） ,we detect the change of oocyte mature rate, Ca2＋ distribution, apoptosis rates, the mitochondrial membrane potential,Bcl-2 and Bax gene expression,this study lay the foundation for the further research that is the influence of heavy metal to female mammals germ cell development and cell apoptosis mechanisms. Bovine oocytes were treated with different concentration of CdC12 （0,0.8,1.6,2.4 μg/mL） for 22 h,we count the rate of maturity and apoptosis;The early apoptosis of bovine ooeytes was detected by Annexin V-FITC;The change of Ca2＋ distribution and the mitochondrial membrane potential were respectively detected by Fluo-3/AM and Rh123 ;Using RT-PCR,we de- tected the relative expression of Bcl-2 and Bax. The mature rate and apoptosis rate of the bovine oocytes by 0.8 g/mL treatment group compared with control group was not significant （ P 〉 0.05 ） ; 1.6 μg/mL and 2.4 μg/mL treatment group compared with control group were not significant （ P 〈 0.05 ） ;with the increase of concentration of CdC12 ,Ca2＋ fluorescence intensity in cytoplasm was increasing,the mitochondrial membrane potential gradually declined,Bcl-2 and Bax Relative ratio was descending. The induction of oocytes apoptos is by CdC12 is dose effect, which can restrain oocytes mature and promote cell apoptosis.

关 键 词：CDCL2 牛卵母细胞 体外成熟 凋亡 

分 类 号：S823[农业科学—畜牧学]