玉米赤霉烯酮对不同时期小鼠子宫细胞凋亡及凋亡因子的影响  

Effects of Zearalenone on Uterine Apoptosis and Apoptosis Factors of Mice in Different Periods

作　　者：王丙雷 王飞[1] 周令强 陈宝江[2] 崔亚利[1] 崔嘉[2] WANG Binglei;WANG Fei;ZHOU Lingqiang;CHEN Baojiang;CUI Yali;CUI Jia(College of Veterinary Medicine,Agricultural University of Hebei,Baoding 071001,China;College of Animal Science and Technology,Agricultural University of Hebei,Baoding 071001,China)

机构地区：[1]河北农业大学动物医学院,保定071000 [2]河北农业大学动物科技学院,保定071000

出　　处：《动物营养学报》2019年第5期2388-2396,共9页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：中国-欧盟饲料霉菌毒素生物脱毒关键技术合作研究项目(S2016G4513)

摘　　要：本试验旨在研究在饲粮中添加不同水平的玉米赤霉烯酮(ZEA)对发情前期、发情期、产仔后雌性小鼠子宫细胞凋亡及相关因子的影响。选取4周龄昆明雌性小鼠120只,随机分为4组,每组30只。选取雄性小鼠20只,随机分为4组,每组5只。7 d预试期结束后,试验分2个阶段。第1阶段,各组雌鼠分别饲喂在基础饲粮中添加0(对照)、50、100、150 mg/kg ZEA的饲粮,饲喂时间是20 d。饲养试验结束后的第2、3、4天,通过阴道涂片法每组挑选出发情前期、发情期小鼠各10只,脱椎处死,摘取子宫角,用于子宫细胞凋亡原位末端标记法(TUNEL)染色和凋亡相关因子mRNA表达量的检测。第2阶段,每组剩余雌鼠与雄鼠按2∶1进行合笼,至产仔后,按上述方法采样检测。结果表明:1)发情前期凋亡细胞主要位于子宫内膜间质细胞及上皮细胞;与对照组相比,添加100、150 mg/kg ZEA极显著提高凋亡阳性表达细胞平均光密度值(P<0.01),且极显著上调半胱天冬蛋白酶(caspase)-3、caspase-8、caspase-9及B细胞淋巴瘤/白血病-2(Bcl-2) mRNA相对表达量(P<0.01); 2)发情期细胞凋亡主要集中在子宫内膜间质细胞、子宫内膜上皮细胞及子宫腺上皮细胞;与对照组相比,随着ZEA添加水平的增加,凋亡细胞呈剂量依赖性增加(P<0.01),能够极显著上调凋亡相关基因mRNA相对表达量(P<0.01); 3)产仔后期凋亡细胞分布较为均匀;与对照组相比,添加150 mg/kg ZEA凋亡程度极显著增加(P<0.01);添加100 mg/kg ZEA能够显著上调Bcl-2、caspase-3 mRNA相对表达量(P<0.05),添加150 mg/kg ZEA能够显著上调caspase-8、caspase-9 mRNA相对表达量(P<0.05)。综上所述,ZEA能够促进不同时期小鼠子宫细胞凋亡,并且能够上调凋亡相关基因caspase-3、caspase-8、caspase-9、Bcl-2表达。The purpose of this study was to investigate the effects of different levels of zearalenone(ZEA)on the apoptosis of uterine cells and related factors in female mice in pre-estrus,estrus and post-partition periods.One hundred and twenty,4-week-old,female Kunming mice were randomly divided into 4 groups,30 in each group.Twenty male mice were randomly divided into 4 groups,5 in each group.After 7 days of pre-trial period,the experiment was divided into two stages.Stage 1:the mice of the four groups were fed the basal diets with 0,50,100 and 150 mg/kg ZEA for 20 days.At days 2,3 and 4 after the feeding test,ten mice in pre-estrus and estrus periods in each group were selected with vaginal smear.The selected mice were sacrificed by cervical delocation,and the uterine horns were harvested for TUNEL staining of apoptotic uterine cells and detection of apoptotic related factors mRNA expression.The remaining females of each group were caged with male mice by 2∶1.After giving birth,the samples were collected according to the above method.The results showed as follows:1)apoptotic cells in pre-estrus period were mainly located in the endometrial stroma and epithelial cells.Compared with the control group,adding 100,150 mg/kg ZEA significantly increased the optical density of apoptotic cells(P<0.01),and significantly increased mRNA relative expression levels of the caspase-3,caspase-8,caspase-9 and B-cell lymphoma/leukemia-2(Bcl-2)(P<0.01).2)In estrous period,apoptosis was observed mainly in endometrial stromal cells,endometrial epithelial cells and uterine gland cells;compared with the control group,with the increasing of ZEA level,apoptotic cells significantly increased(P<0.01),and the mRNA relative expression levels of apoptotic genes significantly increased(P<0.01).3)The distribution of apoptotic cells were more uniform in the post-partition period;compared with the control group,adding 150 mg/kg ZEA significantly increased apoptosis(P<0.01).Meanwhile,adding 100 mg/kg ZEA significantly up-regulated the mRNA relative exp

关 键 词：玉米赤霉烯酮 细胞凋亡 caspase-3 CASPASE-8 CASPASE-9 Bcl-2 

分 类 号：S859.8[农业科学—临床兽医学]