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目的研究亚砷酸钠(Na As O2)暴露对雌雄ICR胎鼠鼠肺Ⅱ型上皮细胞(AECⅡ)雌激素受体α(ERα)表达的影响。方法孕期20 d的ICR胎鼠,分辨雌雄,以Ⅰ型胶原酶分离AECⅡ,以免疫黏附法和差速贴壁法纯化细胞,以免疫荧光法鉴定纯度;以0.5、1.25、5μmol/L Na As O2分别处理AECⅡ12 h和24 h;以不含Na As O2的培养基处理AECⅡ12 h和24 h作空白对照组,每组分设6个重复剂量。以实时荧光定量PCR检测各组ACEⅡ内ERαm RNA表达情况,以western blot检测各组ERα蛋白的表达情况。结果免疫荧光显示纯化后AECⅡ纯度为85%±5%;经Na As O2处理12 h,雌性AECⅡ中各剂量组ERαm RNA和中、高剂量组的蛋白表达高于同剂量组雄性(P<0.05);雌性各剂量组ERαm RNA和蛋白表达均高于同性别对照组;雄性中剂量组蛋白表达高于雄性对照组。经Na As O2处理24 h,雌性低剂量组ERαm RNA表达和中、高剂量组蛋白表达高于同剂量组雄性(P<0.05);雌雄各剂量组ERαm RNA和中高剂量组蛋白表达均高于各自对照组,雌雄对照组之间ERαm RNA和蛋白表达差异无统计学意义(P>0.05)。结论亚砷酸钠暴露可致雌性胎鼠AECⅡ内ERα表达高于雄性。
Objective To investigate the effects of sodium arsenite (Na As O2) exposure on the expression of estrogen receptor α (ERα) in the lung type Ⅱ epithelial cells (AECⅡ) in male and female ICR mice. Methods ICR embryos of 20 d pregnant women were differentiated into female and male ones. AEC Ⅱ was separated by type Ⅰ collagenase, and purified by immunoadsorption and differential adherent methods. The purity of the cells was identified by immunofluorescence. The IC50 of 0.5, 1.25, 5 μmol / L Na As O2 for 12 h and 24 h, respectively. AECⅡ12 h and 24 h were treated with medium without Na As O2 for blank control group. Each group consisted of 6 repeated doses. The expression of ERαmRNA in ACEⅡ in each group was detected by real-time fluorescence quantitative PCR, and the expression of ERα protein in each group was detected by western blot. Results The purity of AECⅡwas 85% ± 5% after immunofluorescence. The protein expression of ERαm RNA and medium and high dose groups of female AECⅡgroup was higher than that of the same dose group (P <0.05) ). The ERαm RNA and protein expression of each dose group were higher than that of the same sex control group; the male middle dose group protein expression was higher than the male control group. The expression of ERαm RNA and the expression of ERαm RNA in female low dose group were higher than those in the same dosage group (P <0.05) by Na As O2 treatment for 24 h. The protein expression of ERαm RNA and medium and high dose group were higher There was no significant difference in ERαm RNA and protein expression between the control group and the male and female controls (P> 0.05). Conclusion Exposure of sodium arsenite induced higher expression of ERα in AECⅡ than that in male.