长春新碱、更生霉素对发育未成熟睾丸影响的实验研究

Influence of vincristine and dactinomycin on spermatogenetic cells of testes in juvenile rats

目的 探讨长春新碱、更生霉素对幼年期大鼠生精细胞的影响。方法 3周龄雄性大鼠根据药物种类及剂量随机分为长春新碱组 (VCR)、更生霉素组 (AMD)、长春新碱 +更生霉素组(VDG)、小剂量长春新碱 +更生霉素组 (sVDG)、对照组。应用HE染色、荧光显微镜、TUNEL技术、免疫组化方法检测大鼠用药后生精细胞凋亡及Fas基因的表达。结果 各实验组睾丸曲细精管管腔内及近管腔生精上皮部分精原细胞显示显著凋亡 (P <0 .0 1) ,VDG生精细胞凋亡峰值分别显著高于VG、DG及sVDG(P >0 .0 5 )。睾丸组织Fas蛋白表达水平在实验组无显著上调。结论 长春新碱与更生霉素都将导致未成熟睾丸精原细胞显著凋亡 ,并与用药种类多少、剂量呈正相关 。

Objective To investigate the effect of vincristine, dactinomycin and its combination on apoptosis of spermatogenic cells in juvenile rats and the expression of Fas gene. Methods 3-week old male rats were randomly divided into 5 groups, namely: vincristine group(VG), dactinomycin group (DG), vincristine plus dactinomycin group(VDG),smaller-dose vincristine and dactinomycin group (sVDG) and control group(CG). The rats of the former 4 vincristine and dactinomycin group(VDG) had intraperitoneal injection of the drugs. HE stain, fluorescent microscope, TUNEL and immunohistochemical stain were used to detect the apoptotic response of spermatogenic cells to vincristine, dactinomycin and its combination, and the expression of Fas gene on 12h and day 1, 2, 3, 4, 6 after the first injection. Results A few spermatogenic cells were seen near or in the lumen of the seminiferous tubules, which had highly condensed cytoplasm with bacilliform nucleus or nuclear fragments in all treatment groups. Apoptosis of spermato-genetic cells were found in all therapy groups as compared to CG by TUNEL (P< 0.05). Apototic peaks of VG, DG, VDG, sVDG occurred on day 1, 2, 3, after the first injection respectively. The incidence of apoptotic germ cells of VDG was higher than VG, DG and sVDG. (P< 0.01). No increase of Fas protein were observed after injection of drugs (P> 0.05). Conclusions Vincristine and dactinomycin leads apoptotic to spermatogonia in juvenile rats. The study demonstrates that it is important to follow up the fertility of children with Wilms' tumor who ever received vincristine and dactinomycin therapy.