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目的探讨抗肿瘤药物所致生精细胞损害与年龄的关系,为性腺保护剂的运用寻找理论依据。方法将环磷酰胺分别作用于处于不同发育时期的1周龄、3周龄、5周龄、9周龄雄性大鼠,应用HE染色法、TUNEL法和免疫组化法检测急性期生精细胞凋亡,bcl-2蛋白表达,细胞增殖能力变化及远期组织学损害。结果用药后24h,除1周龄组外,各实验组生精细胞显著凋亡(P<0.01),bcl-2蛋白表达显著下降(P<0.01),生精细胞S期所占的比例显著下降(P<0.01)。用药后9周,除1周龄组外,各实验组曲细精管面积、直径、生精上皮细胞计数、Johnsen’s评分均显著低于相应对照组(P<0.01),并随年龄增大损害有增加趋势。结论环磷酰胺可诱导生精细胞增殖启动阶段以后的生精细胞显著凋亡,且伴有bcl-2明显下调。并可显著抑制精原细胞和细线前期精母细胞增殖。远期组织学改变年龄越小所受的损害越小,提示为使睾丸免受抗肿瘤药物的伤害而采用性激素使生精细胞增殖分化处于不分化状态是有效的防护方法之一。
Objective To investigate the relationship between the damage of spermatogenic cells induced by antitumor drugs and age, and to find the theoretical basis for the application of gonadotropin. Methods Cyclophosphamide was administered to male rats at 1 week, 3 weeks, 5 weeks and 9 weeks at different developmental stages. HE staining, TUNEL and immunohistochemistry were used to detect the apoptosis of spermatogenic cells Bcl-2 protein expression, cell proliferation ability and long-term histological damage. Results The apoptosis of spermatogenic cells (P <0.01) and the expression of bcl-2 in experimental group were significantly decreased (P <0.01) except for 1-week group at 24 h after treatment. The proportion of S phase of spermatogenic cells was significantly Decreased (P <0.01). At 9 weeks after administration, except the 1-week-old group, the seminiferous tubule area, diameter, spermatogenic epithelial cell count and Johnsen’s score in each experimental group were significantly lower than those in the corresponding control group (P <0.01) Increase the trend. Conclusion Cyclophosphamide can induce significant apoptosis of spermatogenic cells after the initiation of spermatogenic cell proliferation, accompanied by significant down-regulation of bcl-2. And can significantly inhibit the proliferation of spermatogonia and early spermatocytes. Long-term histological changes in the smaller the younger the damage suffered, suggesting that the testes from the antitumor drugs and the use of sex hormones spermatogenic cells proliferation and differentiation in an undifferentiated state is an effective protective method.