丙烯腈对大鼠睾丸中雄激素结合蛋白和抑制素基因表达的影响

Effects of Acrylonitrile on Gene Expression of Androgen Binding Protein and Inhibin in Testes of Spraque-Dawley Rats

[目的]研究丙烯腈(acrylonitrile,ACN)对大鼠睾丸中雄激素结合蛋白(Androgen Binding Protein,ABP)和抑制素基因表达的影响。[方法]SD雄性大鼠随机分为4组,1组为对照组,另3组为染毒组,剂量分别为7.5、15.0和30.0 mg/kg体重,每天腹腔注射染毒1次,每周5次,连续13周,分别于4、8和13周末分批处死,取睾丸组织50-100 mg, 加1 ml Trizol磨成匀浆,加氯仿振摇,12 000 r/min,4℃下离心,取上清液,加异丙醇沉淀,75%乙醇洗涤,得到的RNA用紫外分光光度法测定其纯度,用一步法逆转录扩增(reverse transcription-polymerase chain reaction,RT-PCR)目的基因,用内参标进行半定量检测。[结果]ACN染毒未引起ABP和抑制素基因表达缺失。低、中剂量组睾丸中ABP和抑制素基因表达水平无明显变化。高剂量组,染毒4周后,ABP基因表达水平显著升高(P<0.05),染毒8周后下降,但与其他组无明显差异,13周后显著下降,与其他组有显著性差异(P<0.05)。高剂量组染毒4周后,抑制素基因表达水平有下降趋势,且随染毒时间延长下降显著;中、高组在染毒8周后,抑制素基因表达水平下降显著,与对照组比较差异有显著性(P<0.01)。[结论]ACN对大鼠睾丸中ABP和抑制素基因表达水平有影响,提示丙烯腈对睾丸中支持细胞可能有损伤作用。

[ Objective ] The purpose of this paper was to study effects of acrylonitrile （ ACN ）on gene expression of androgen binding protein（ ABP ） and inhibin in testes of rats. [ Methods ] Male Spraque-Dawley（ SD ） rats were randomly divided into four groups, one group as a control group, other three groups were daily administrated ACN by intraperitoneal injection at the doses of 7.5, 15.0 and 30.0 mg/kg body weight 5 times a week for 13 weeks, respectively. Experimental rats were sacrificed and testes were removed at the end of 4, 8, 13 weeks, respectively. 50-100 mg of testicle tissue was taken and ground into homogeneous liquid with lml trizol. RNA was extracted, separated and purified by normal method. The pureness of RNA was measured by ultraviolet spectrometry. Objective gene was amplified with reverse transcription-polymerase chain reaction （ RT-PCR ）.Gene expression levels were determined semi-quantitatively by inner reference standard. [ Results ] No loss occurred in gene expression of ABP and inhibin induced by ACN. There was no significant difference in gone expression levels of ABP and inhibin in the testes induced by ACN at and below the dose level of 30 mg/kg body weight. After treatment for 4 weeks, ACN caused significant increase in level of ABP gene expression at the dose level of 30.0 mg/kg body weight（ P 〈 0.05 ） .There was no significant difference among all groups in levels of ABP gene expression after treatment for 8 weeks. Levels of ABP gene expression were significantly decreased in 30.0 mg/kg body weight group after treatment for 13 weeks（ P 〈 0.05 ）.There was no significant difference in level of inhibin gone expression following 4-week treatment among all groups, then a tendency to decrease occurred with longer treatment duration. The level of inhibin gene expression was significantly decreased at the dose of ACN ≥ 15 mg/kg following 8-week treatment （ P 〈 0.01 ）. [ Conclusion ] The results indicate that ACN could induce changes in the levels of ABP and inhibin gene expression in testes of male SD rats, that may damage Sertoli cells in testes.