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目的通过观察不同剂量二甲磺酸乙烷(EDS)对成年大鼠Leydig细胞的杀伤效应,确定EDS的最佳剂量。方法6个月龄SD成年雄性大鼠42只,随机分为EDS处理组、溶酶对照组和正常对照组,其中EDS处理组按照剂量的不同设置40、60、75、90和130 mg/kg体重5个亚组,每个亚组含6只大鼠。腹腔内注射给药,注射EDS后3、7和14d处死大鼠,取出睾丸组织,行苏木素-伊红(HE)染色光镜下组织学观察和P450scc免疫组织化学观察及灰度分析。结果正常对照和溶酶对照组Leydig细胞形态、数目正常,曲细精管内各级生精细胞排列有序,无紊乱现象。EDS处理3d后,130 mg/kg体重剂量组的2只大鼠死亡,其他剂量组均观察到Leydig细胞的减少,其中40 mg/kg体重剂量组的减少程度不明显;EDS处理后7d,曲细精管内生精细胞出现排列紊乱的现象,以90 mg/kg体重剂量组更为明显;EDS处理后14d,60 mg/kg体重剂量组出现一种核圆形、核染色淡、体积较大的新形成Leydjg细胞,而在75 mg/kg体重剂量组没有发现此细胞。P450scc免疫组织化学与光镜结果一致。灰度分析结果显示,EDS处理后3及7 d,各剂量组与正常对照比较差异均有统计学意义(P<0.05);60 mg/kg体重剂量组14与3、7d比较差异有统计学意义(P<0.05),且与正常对照差异无统计学意义(P>0.05)。结论EDS确实能杀死成年大鼠Leydig细胞,60 mg/kg体重的剂量可以更好的模拟青春期Leydig细胞增殖分化过程。
OBJECTIVE To determine the optimal dose of EDS by observing the killing effect of different doses of ethanedimethanesulfonate (EDS) on adult rat Leydig cells. Methods Forty-two SD adult male rats of 6 months old were randomly divided into EDS treatment group, lysozyme control group and normal control group. The EDS treatment groups were given 40, 60, 75, 90 and 130 mg / kg Body weight 5 subgroups, each subgroup containing 6 rats. The rats were sacrificed at 3, 7 and 14 days after injection of EDS. The testicular tissues were removed and stained with hematoxylin and eosin (HE) for histological observation and immunohistochemical staining and gray scale analysis of P450scc. Results The number of normal and control group Leydig cells were normal and the number of spermatogenic cells in seminiferous tubules were arranged orderly and without disorder. Two days after EDS treatment, two rats died at the dose of 130 mg / kg body weight, and Leydig cells decreased at the other dose groups, but the reduction was not obvious at the dose of 40 mg / kg body weight. At 7 days after EDS treatment, The spermatids of seminiferous tubules were disordered, which was more pronounced at the dose of 90 mg / kg body weight. At 14 days after EDS treatment, a nucleus was found in the dose of 60 mg / kg body weight, Of the newly formed Leydjg cells, whereas no such cells were found at the 75 mg / kg body weight dose. P450scc immunohistochemistry and light microscopy results. The results of grayscale analysis showed that there was significant difference between each dose group and the normal control at 3 and 7 d after EDS treatment (P <0.05), and there was statistical difference between 14 and 3 and 7 d of 60 mg / kg body weight dose group Significance (P <0.05), and there was no significant difference with the normal control (P> 0.05). Conclusion EDS can kill adult Leydig cells. The dose of 60 mg / kg body weight can better simulate the process of adolescent Leydig cell proliferation and differentiation.