砷对大鼠雌激素受体基因及其下游细胞因子表达的影响  

Effects of arsenite on the expression of estrogen receptor gene and the ER signaling pathway in rats

作　　者：郑冲[1] 张人华[1] 

机构地区：[1]贵州省疾病预防控制中心卫生监测检验所,贵阳550004

出　　处：《中华地方病学杂志》2016年第10期731-735,共5页Chinese Journal of Endemiology

基　　金：贵州省疾病预防控制中心青年技术基金（2014-E1-5青）

摘　　要：目的 探讨砷对大鼠雌激素受体（estrogen receptor,ER）基因及其下游细胞因子表达的影响.方法 24只雌性SD大鼠,采用随机数字表法分成4组,分别为对照组、低砷组（Aslow）、中砷组（Asmiddle）、高砷组（Ashigh）,每组6只大鼠.AslowAsmiddle、Ashigh组采用亚砷酸钠（NaAsO2）灌胃染毒,剂量分别为0.625、2.500、10.000mg/kg.灌胃染毒3个月,提取大鼠卵巢组织总RNA,实时荧光定量PCR（RT-PCR）法检测ER mRNA的表达水平;酶联免疫吸附法（Enzyme-linked immunosorbnent assay,ELISA）检测大鼠血清中ER、血管内皮生长因子（vascular endothelial growth factor,VEGF）、细胞周期素D1 （cyclin D1,CYC-D1）、孕激素受体（progesterone receptor,PR）的表达水平.结果 对照组、Aslow Asmiddle、Ashigh组间ER mRNA,血清VEGF、PR、CYC-D1、ER表达水平比较差异均有统计学意义（F=204.13、227.14、256.13、179.23、167.34,P均＜0.01）.随着砷剂量的增高,ER mRNA,血清VEGF、PR表达水平逐渐降低（P均＜0.01）,剂量-效应关系明显;而血清CYC-D1、ER表达水平在Asmiddle、Ashigh组间比较差异均有统计学意义（P均＜ 0.01）,Asmiddle组[（25.45±9.29）U/L,（22.16±9.16）ng/L]高于Ashigh组[（2.60±0.23）U/L,（9.06±2.06）ng/L].染砷含量、尿砷（UAs）与ER mRNA,血清ER、VEGF、CYC-D1、PR表达水平均呈负相关关系（r=-0.689、-0.515、-0.675、-0.456、-0.397,P＜0.01或＜0.05）.ER mRNA与血清ER、VEGF、CYC-D1、PR表达水平均呈正相关关系（r=0.894、0.662、0.713、0.699,P＜0.01或＜0.05）.结论 砷抑制大鼠卵巢ER mRNA及血清ER蛋白表达,ER可作为评价砷暴露生殖损伤的分子生物学标志;ER-VEGF、CYC-D1、PR信号通路参与了砷暴露致大鼠生殖系统的损伤,砷很可能是环境雌激素类物质.Objective To explore the effects of arsenite on the expression of estrogen receptor （ER） mRNA,ER protein and the ER signaling pathway in rats.Methods Random grouping experiment design was used to evaluate the effects of arsenite on the expression of ER mRNA,ER protein and the ER signaling pathway （24 rats were divided into 4 groups：control,Aslow,Asmiddle,Ashigh groups;6 rats in each group）.Female rats were exposed to NaAsO2 （0.625,2.500,10.000 mg/kg） by oral perfusion at gradient doses for three months respectively.The expression of ER mRNA as effect biomarker was observed through RT-PCR.The concentration of ER,vascular endothelial growth factor （VEGF）,cyclin D1 （CYC-D1）,and progesterone receptor （PR） in serum was determined as effect biomarkers using enzyme-linked immunosorbent assay （ELISA）.Results The levels of ER mRNA,VEGF,PR,CYC-D1,and ER in control,Aslow,Asmiddle,and Ashigh groups were statistically different （F =204.13,227.14,256.13,179.23,167.34,all P 〈 0.01）.With increasing of arsenic,the expression of ER mRNA,VEGF,and PR gradually decreased （all P 〈 0.01）;the dose effect relationship was obvious;the expression of CYC-D1 and ER in Asmiddle [（25.45 ± 9.29） U/L,（22.16 ± 9.16） ng/L] was higher than that in Ashigh [（2.60 ± 0.23） U/L,（9.06 t 2.06） ng,/L,all P 〈 0.01].Arsenic was negatively associated with ER mRNA,ER,VEGF,CYC-D1 and PR （r =-0.689,-0.515,-0.675,-0.456,-0.397,P 〈 0.01 or 〈 0.05）.ER mRNA was positively associated with ER,VEGF,CYC-D1 and PR （r =0.894,0.662,0.713,0.699,P 〈 0.01 or 〈 0.05）.Conclusions The expression of ER mRNA and ER protein are inhibited by arsenic,ER may be served as a sensitive biomarker of reproductive system damage after exposed to arsenic.The ER-VEGF,CYC-D1 and PR pathway participate in the process of reproductive system damage caused by arsenic.Arsenic is probably a environmental estrogen.

关 键 词：砷 雌激素受体 血管内皮生长因子 细胞周期素D1 孕激素受体 

分 类 号：R599.1[医药卫生—内科学]