壬基酚对大鼠垂体和N2a细胞雌激素受体β表达的影响  

Effects of nonylphenol on estrogen receptorβexpression in rat pituitary and N2a cells

作　　者：李凤 张钰婕 李胜男 罗娅 俞捷 许洁 LI Feng;ZHANG Yu-jie;LI Sheng-nan;LUO Ya;YU Jie;XU Jie(School of Public Health,Zunyi Medical University,Zunyi,Guizhou 563000,China)

机构地区：[1]遵义医科大学公共卫生学院,贵州遵义563000

出　　处：《现代预防医学》2023年第11期2076-2081,共6页Modern Preventive Medicine

基　　金：国家自然科学基金(22166035,22266039);贵州省高层次创新型“百层次”人才基金(黔科合平台人才[2020]6014,黔科合平台人才-GCC[2022]035-1);遵义市“15851人才”项目经费(2019-E262)。

摘　　要：目的探讨壬基酚(nonylphenol,NP)对大鼠垂体组织及神经瘤母细胞(neuro-2a cells,N2a)中雌激素受体β(estrogen receptorβ,ERβ)表达水平的影响。方法(1)24只SD雄性大鼠随机分成4组,每组6只,分别为空白对照组(C,玉米油),NP低剂量组(L-NP,0.4 mg/kg)、中剂量组(M-NP,4 mg/kg)、高剂量组(H-NP,40 mg/kg),空腹灌胃90 d后进行悬尾实验;(2)取大鼠垂体组织,高效液相色谱测定NP含量、免疫荧光检测ERβ的表达水平;(3)0~200μmol/L NP细胞染毒后,显微镜下观察细胞,确定细胞实验的NP剂量;CCK-8法检测剂量浓度NP影响下N2a细胞的吸光度,免疫荧光检测N2a细胞ERβ表达。结果壬基酚可致大鼠不动时间延长(F=11.28,P<0.01)且随着染毒剂量的增加而增加。与对照组相比,各NP染毒组垂体ERβ荧光强度变暗(F=6.335,P<0.05);垂体NP含量与不动时间呈正相关(r=0.642,P<0.01)、与垂体ERβ荧光表达呈负相关(r=-0.984,P<0.05)。0~200μmol/L NP细胞染毒剂量与细胞存活率呈负相关(r=-0.963,P<0.01);从40μmol/L开始,随着染毒剂量的增加,细胞存活率下降(P<0.01),N2a细胞形态学观察显示,细胞透光性变差、贴壁不好、细胞形态发生改变;细胞ERβ免疫荧光强度变弱。结论壬基酚可在大鼠垂体蓄积、降低垂体ERβ表达水平,具有神经细胞毒性。Objective To investigate the effect of nonylphenol(NP)on the expression of estrogen receptorβ(ERβ)in rat pituitary and Neuro-2a cells(N2a).Methods(1)24 male SD rats were randomly divided into 4 groups with 6 rats in each group:blank control group(C,corn oil),low dose NP group(0.4 mg/kg),medium dose group(4 mg/kg)and high dose group(40 mg/kg).The tail suspension test was performed after 90 days of fasting.(2)The content of NP in rat pituitary tissue was determined by high performance liquid chromatography and the expression of ERβwas detected by immunofluorescence.(3)After 0 to 200μMNP cells were exposed,the cells were observed under microscope to determine the dose of NP,the absorbance of N2a cells was detected by CCK-8 method,and the expression of ERβin N2a cells was detected by immunofluorescence.Results Nonylphenol could prolong the immobility time of rats(F=11.28,P<0.01)and increased with the increase of dose.Compared with the control group,the fluorescence intensity of ERβin pituitary of all NP exposed groups was darker(P<0.05),and the content of NP in pituitary was positively correlated with immobility time(P<0.01)and negatively correlated with the fluorescence expression of ERβin pituitary(r=0.984,P<0.05).There was a negative correlation between the dose of 0 to 200μMNP cells and the cell survival rate(r=0.963,P<0.0l),and the cell survival rate decreased with the increase of exposure dose from 40μM.The morphological observation of N2a cells showed that the light transmittance of N2a cells became worse,the adhesion was not good,the cell morphology changed,and the immunofluorescence intensity of ERβdecreased.Conclusion Nonylphenol can accumulate in rat pituitary and reduce the expression level of ERβin pituitary.Nonylphenol has neurotoxicity.

关 键 词：壬基酚 N2A细胞 雌激素受体Β 

分 类 号：R114[医药卫生—卫生毒理学]