【摘要】：目的:評價細(xì)胞毒性藥物二甲磺酸乙烷對新生大鼠睪丸胚胎型Leydig細(xì)胞的作用效應(yīng)及安全使用劑量。方法:選取出生后3 d的雄性SD幼鼠共40只,分為對照組(二甲基亞砜溶劑組)、二甲磺酸乙烷75 mg/kg組、二甲磺酸乙烷100 mg/kg組和二甲磺酸乙烷125 mg/kg組,每組10只。予腹腔內(nèi)注射不同實驗劑量的二甲磺酸乙烷,注射后4 d分別處死幼鼠;測量動物體重及睪丸重量,放射免疫法檢測血清中睪酮水平,睪丸組織冰凍切片行3β-羥基類固醇脫氫酶(3β-hydroxysteroid dehydrogenase,3β-HSD)染色,實時熒光定量PCR技術(shù)及Western blot技術(shù)檢測睪丸組織中Leydig細(xì)胞相關(guān)特異基因及蛋白的表達(dá)水平。結(jié)果:藥物注射后4 d,與對照組相比較,在血清睪酮水平方面[(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劑量組輕度下降,而100 mg/kg劑量組和125 mg/kg劑量組則明顯降低;睪丸組織3β-HSD染色顯示:75 mg/kg劑量組仍可見少量的3β-HSD染色呈陽性的胚胎型Leydig細(xì)胞(fetal leydig cell,FLC),100 mg/kg劑量組和125 mg/kg劑量組無3β-HSD染色呈陽性的FLC存在,但125 mg/kg劑量組睪丸曲細(xì)精管細(xì)胞排列嚴(yán)重紊亂;RT-PCR及Western blot檢測顯示:100 mg/kg劑量組睪丸組織Hsd3b1和Cyp17a1的mRNA表達(dá)水平及其相應(yīng)蛋白表達(dá)水平明顯降低(P0.001)。結(jié)論:二甲磺酸乙烷能夠特異性地破壞新生雄性SD大鼠睪丸組織中的胚胎型Leydig細(xì)胞,腹腔注射100 mg/kg二甲磺酸乙烷可建立起穩(wěn)定、可靠的大鼠睪丸去胚胎型Leydig細(xì)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 鹵0.117) 渭 g 鈮，

本文編號：2515913