摘要
研究了壬基酚异构体[4-(1-乙基-1-甲基-己基)酚](4-NP65)对小鼠睾丸Sertoli TM4细胞黏着连接相关分子表达和分泌雄激素结合蛋白(ABP),抑制素B(Inhibin B)的作用。不同浓度4-NP65(0、0.1、1、10、20、40μmol·L-1)作用于TM4细胞24h,MTT法测定细胞存活率,RT-PCR测定TM4细胞波形蛋白(Vimentin),N-钙黏着蛋白(N-cadherin),黏着斑蛋白(Vinculin)mRNA的表达情况;Elisa法测定细胞上清液中ABP和Inhibin B的含量。结果4-NP65在低剂量(0.1、1、10、20μmol·L-1)对细胞存活率无显著影响,在高剂量(40、80、100μmol·L-1)能显著降低TM4细胞存活率(P<0.01);4-NP65剂量依赖性增加Vimentin mRNA表达,降低Vinculin、N-cadherin mRNA的表达;4-NP65呈剂量依赖性降低ABP的分泌量,与对照组比较,在20、40μmol·L-1处理组有显著差异(P<0.05),而Inhibin B的含量在20、40μmol·L-1 4-NP65处理组有显著降低(P<0.05)。4-NP65对小鼠Sertoli TM4细胞具有生殖毒性作用,黏着连接是其作用的可能靶点之一,4-NP65可能通过影响TM4细胞黏着连接相关分子的表达和TM4细胞分泌ABP、Inhibin B引起雄性生殖系统损伤。
The aim of this study was to investigate the effects of nonylphenol isomer（4-NP65）exposure on the expression of adherens junction-associated molecules and secretion of ABP,Inhibin B in mouse Sertoli TM4 cells.Cultured cells were treated with 4-NP65（0,0.1,1,10,20,40μmol·L-1）for 24h,then cell survival rate was detected by MTT method;the expression of Vimentin,N-cadherin,Vinculin mRNA was analyzed by RT-PCR and the contents of ABP,Inhibin B in the supernatant were examined by Elisa assay.Compared with Control group,no significant changes have been observed in 0.1,1,10,or 20μmol·L-1,however,4-NP65reduced the cell survival rate significantly at 40,80 or 100μmol·L-1.Additionally,4-NP65 upregulated Vimentin mRNA levels in a dose dependent manner,on contrary,4-NP65 reduced N-cadherin,Vinculin mRNA expression.The ABP content decreased significantly in a dose dependent manner.In addition,the Inhibin B levels decreased dramaticly in 20,40μmol·L-1 4-NP65groups.These ndings demonstrated that 4-NP65 could exert a certain toxicity in TM4cells,4-NP65 treatment may cause the reproductive dysfunction in male via influence expression of adherens junction-associated molecules and secretion of ABP,Inhibin B in mouse Sertoli TM4cells.
出处
《南昌大学学报（理科版）》
CAS
北大核心
2013年第5期469-473,共5页
Journal of Nanchang University(Natural Science)
基金
国家"973"计划项目(2012CB720805)
科技部国际合作项目(2010DFA31780)
国家基金委中德科学基金中心合作项目(GZ731)
