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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。
Twenty-four rabbits were randomly divided into control group and experimental group Ⅰ, Ⅱ, and Ⅲ. The diets of the experimental group were respectively treated with 15% of the first group (30mg / kg of sirolimus), 30% of the second group 60mg / kg) and 45% (wladimin 90mg / kg) in group Ⅲ, respectively. The control group was only fed with hay, the test period was 70 days. The testis of two rabbits was collected randomly from each group. The apoptosis was detected by TUNEL method. The expression of Bcl-2 and Bax mRNA was detected by real-time PCR and the expression of Bcl-2 and Bax protein by immunohistochemistry. The results showed that from the 35th day, the apoptotic index, Bcl-2 and Bax mRNA expression in test group Ⅱ and group Ⅲ were significantly different from those in control group (P <0.01). In test group Ⅰ, the testicular cells (P <0.05). The Bcl-2 protein expression in testis tissue of test group was significantly lower than that of control group, while the expression of Bax protein was significantly higher than that of control group Differences with the extension of poisoning time changes. The results showed that hypoxia poisoning could induce the apoptosis of spermatogenic cells in testis tissue of rabbits, and had a time-dose effect on Oxytropis halophila. This effect may be related to the gene expression of Bcl-2 and Bax.