小花棘豆中毒对家兔睾丸细胞凋亡及Bcl-2、Bax表达的影响  

Effects of oxytropis glabra DC poisoning on cells apoptosis and expression of Bcl-2 and Bax in rabbit testis

作　　者：王帅[1] 贾琦珍[1] 矫继峰[1] 陈根元[1] 

机构地区：[1]塔里木大学动物科学学院/新疆生产建设兵团塔里木畜牧科技重点实验室,新疆阿拉尔843300

出　　处：《中国兽医学报》2017年第1期148-153,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31460678;31260586);新疆生产建设兵团塔里木畜牧科技重点实验室开放课题资助项目(HS201409)

摘　　要：将24只家兔随机分为对照组和试验Ⅰ、Ⅱ、Ⅲ组,试验组饲粮分别按照Ⅰ组15%(苦马豆素含量30mg/kg)、Ⅱ组30%(苦马豆素含量60mg/kg)、Ⅲ组45%(苦马豆素含量90mg/kg)的比例添加小花棘豆,对照组仅饲喂青干草,试验期70d;分别于攻毒后第14,35,70天每次每组随机采集2只家兔的睾丸,通过TUNEL法检测细胞凋亡,real-time PCR检测Bcl-2、Bax mRNA的表达,免疫组化检测Bcl-2、Bax蛋白的表达。结果显示:从第35天开始,试验Ⅱ组和Ⅲ组家兔睾丸细胞凋亡指数、Bcl-2和Bax mRNA的表达均与对照差异极显著(P<0.01),试验Ⅰ组家兔睾丸细胞凋亡指数、Bcl-2和Bax mRNA的表达均与对照差异显著(P<0.05);试验组家兔睾丸组织Bcl-2蛋白表达均明显低于对照,Bax蛋白表达均明显高于对照,其差异性随中毒时间的延长而变化。结果表明:小花棘豆中毒可导致家兔睾丸组织生精细胞凋亡,且与小花棘豆中毒呈现一定的时间-剂量效应,这种作用可能与Bcl-2和Bax基因表达有关。The aim of this study was to investigate the Twenley two effects of Oxytropis glabra DC poisoning on the occurrence of cells apoptosis and the expression of the apoptosis related gene Bel-2 and Bax in rabbit testis, and reveal the toxicity mechanism of O. glabra DC in future. Twen- ty four male rabbits were randomly and equally divided into blank control group and experimental group Ⅰ , Ⅱ and Ⅲ（6 rabbits in each group）, the dried plant of O. glabra DC was comminuted. then different amounts of the grass powder （15 %. 30 %and 45 % corresponding to 30,60 and 90 mg/kg swainsonine,respectively） were mixed with the feeds in the three experimental groups. The rabbits consumed the forages freely until typical symptoms were observed, the test period was 70 d. Two rabbits were selected randomly from each group on the 14th. 35th and 70th day,re- spectively,the testis was collected and apoptosis of its cells was measured by in situ end-labeled DNA（TUNEL）, the mRNA expression of Bcl-2 and Bax in different eneephalie regions was meas- ured by real-time polymerase chain reaction（real-time PCR）, the expression level of Bcl-2 and Bax protein was measured by immunohistochemical staining. The expression of Bcl-2 mRNA in experi- mental group Ⅰ , Ⅱ andⅢ were very signifieanly lower than that of control group（P〈0.05 or 0.01） ,and the apoptosis index of testis cell and the expression of Bax mRNA increased in experl-mental group Ⅰ , Ⅱ andⅢ were very significantly lower than control group（P〈0.05 or 0.01）. Compared with control group,the decreased protein expression of Bcl-2 and the increased protein expression of Bax were detected in Ⅰ , Ⅱ andⅢ group. The results showed that O. glabra DC has remarkable effects on the occurrence of cell apoptosis and the expression of the apoptosis related gene Bcl-2 and Bax in rabbit testis,in a time-effect and a dose-effect relationship. The cell apoptot- sis is highly associated with O. glabra DC poisoning and changes in Bcl-2/Bax expression may play

关 键 词：小花棘豆 睾丸 家兔 凋亡 Bcl^2/Bax 

分 类 号：S852.2[农业科学—基础兽医学] S856.9[农业科学—兽医学]