二甲磺酸乙烷对大鼠睾丸胚胎型Leydig细胞的作用

发布时间：2019-07-18 14:02

【摘要】：目的:评价细胞毒性药物二甲磺酸乙烷对新生大鼠睾丸胚胎型Leydig细胞的作用效应及安全使用剂量。方法:选取出生后3 d的雄性SD幼鼠共40只,分为对照组(二甲基亚砜溶剂组)、二甲磺酸乙烷75 mg/kg组、二甲磺酸乙烷100 mg/kg组和二甲磺酸乙烷125 mg/kg组,每组10只。予腹腔内注射不同实验剂量的二甲磺酸乙烷,注射后4 d分别处死幼鼠;测量动物体重及睾丸重量,放射免疫法检测血清中睾酮水平,睾丸组织冰冻切片行3β-羟基类固醇脱氢酶(3β-hydroxysteroid dehydrogenase,3β-HSD)染色,实时荧光定量PCR技术及Western blot技术检测睾丸组织中Leydig细胞相关特异基因及蛋白的表达水平。结果:药物注射后4 d,与对照组相比较,在血清睾酮水平方面[(0.542±0.117)μg/L,(0.124±0.021)μg/L,(0.113±0.015)μg/L,vs.(0.834±0.172)μg/L,P0.05],75 mg/kg剂量组轻度下降,而100 mg/kg剂量组和125 mg/kg剂量组则明显降低;睾丸组织3β-HSD染色显示:75 mg/kg剂量组仍可见少量的3β-HSD染色呈阳性的胚胎型Leydig细胞(fetal leydig cell,FLC),100 mg/kg剂量组和125 mg/kg剂量组无3β-HSD染色呈阳性的FLC存在,但125 mg/kg剂量组睾丸曲细精管细胞排列严重紊乱;RT-PCR及Western blot检测显示:100 mg/kg剂量组睾丸组织Hsd3b1和Cyp17a1的mRNA表达水平及其相应蛋白表达水平明显降低(P0.001)。结论:二甲磺酸乙烷能够特异性地破坏新生雄性SD大鼠睾丸组织中的胚胎型Leydig细胞,腹腔注射100 mg/kg二甲磺酸乙烷可建立起稳定、可靠的大鼠睾丸去胚胎型Leydig细胞模型。
[Abstract]:Aim: to evaluate the effect and safe dose of cytotoxicity drug ethane dimethyl sulfonate on testicular embryonic Leydig cells in neonatal rats. Methods: a total of 40 young male SD rats at 3 days after birth were divided into control group (dimethyl sulfoxide solvent group), dimethyl sulfonate ethane 75 mg/kg group, dimethyl sulfonate ethane 100 mg/kg group and dimethyl sulfonate ethane 125 mg/kg group with 10 rats in each group. The young rats were killed 4 days after intraabdominal injection of different experimental doses of ethane dimethyl sulfonate, the body weight and testicular weight of the animals were measured, the serum testosterone level was measured by radioimmunoassay, the frozen sections of testicular tissue were stained with 3 尾-hydroxysteroid dehydrogenase,3 尾-HSD, and the expression of specific genes and proteins related to Leydig cells in testicular tissue was detected by real-time fluorescence quantitative PCR and Western blot. Results: at 4 days after injection, compared with the control group, the serum testosterone level [(0.542 卤0.117) 渭 g 鈮，

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