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探讨小花棘豆中毒对家兔睾丸生精细胞p53、Bcl-2和Bax基因的影响。将24只公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%、Ⅱ组30%、Ⅲ组45%的比例添加小花棘豆,对照组仅饲喂苜蓿干草,试验期70 d。分别于攻毒后第14天、第35天、第70天每次每组随机采集2只家兔的睾丸,通过Real-time q PCR检测生精细胞p53、Bcl-2和Bax m RNA的表达,免疫组化和Western Blot检测生精细胞p53、Bcl-2和Bax蛋白的表达。结果显示,试验Ⅱ组和Ⅲ组家兔睾丸生精细胞p53、Bcl-2和Bax m RNA的表达均与对照差异极显著(p<0.01),试验Ⅰ组家兔睾丸生精细胞p53、Bcl-2和Bax m RNA的表达均与对照差异显著(p<0.05);试验组家兔生精细胞中Bcl-2蛋白表达均明显低于对照,p53和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相关基因p53、Bcl-2和Bax表达异常,且与小花棘豆中毒呈现一定的时间-剂量效应。
Abstract：The aim of this study was to investigate the effects of Oxytropis glabra DC poisoning on the expression of the apoptosis related gene p53, Bcl-2 and Bax in rabbit testis, and reveal the toxicity mechanism of O. glabra DC in future. 24 male rabbits were randomly and equally divided into blank control group and experimental groupⅠ, Ⅱ, Ⅲ(6 rabbits in each group), the dried plant of O. glabra DC was comminnuted, then different amounts of the grass powder 15%, 30% and 45% 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. 2 rabbits were selected randomly from each group on the 14 th, 35 thand 70thday respectively, the testis were collected and the m RNA expression of p53, Bcl-2 and Bax in different encephalic regions were measured by Real-time polymerase chain quantitative reaction(Real-time q PCR), the level of p53, Bcl-2 and Bax protein expression were measured by immunohistochemical staining and Western Blot. The expression of Bcl-2 m RNA and protein decreased in experimental group Ⅱ, Ⅲ were very significant lower than control group(p<0.01), and the expression of p53, Bax m RNA and protein increased in experimental groupⅡ, Ⅲ were very significant higher than control group(p<0.01), the expression of Bcl-2 m RNA and protein decreased in experimental groupⅠwas significant lower than control group(p<0.05), the expression of p53, Bax m RNA and protein increased in experimental group Ⅰ was significant higherthan control group(p<0.05). The results showed that O. glabra DC has remarkable effects on the occurrence of cell apoptosis and the expression of the apoptosis related gene p53, Bcl-2 and Bax in rabbit testis, in a time-effect as a dose-effect relationship. The apoptotic cell death is highly associated with O. glabra DC poisoning and changes in p53, Bcl-2 and Bax expression may play an important role in this process.
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基本信息:
DOI：10.13417/j.gab.035.001115
中图分类号:S858.291
引用信息:
[1]王帅,贾琦珍,陈根元,等.小花棘豆中毒对家兔生精细胞p53、Bcl-2和Bax表达的影响[J].基因组学与应用生物学,2016,35(05):1115-1122.DOI:10.13417/j.gab.035.001115.
基金信息:
国家自然科学基金(批准号:31460678);; 国家星火计划(批准号:2015GA891015);; 新疆生产建设兵团塔里木畜牧科技重点实验室开放课题(批准号:HS201409)共同资助
2016-05-25
2016-05-25