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杨梦平, 车望军, 程亚龄等. 砷暴露及雌激素受体拮抗剂对雌、雄胎鼠AECⅡ内ERβ表达的影响[J]. 四川大学学报(医学版), 2018, 49(3): 364-368.
引用本文: 杨梦平, 车望军, 程亚龄等. 砷暴露及雌激素受体拮抗剂对雌、雄胎鼠AECⅡ内ERβ表达的影响[J]. 四川大学学报(医学版), 2018, 49(3): 364-368.
YANG Meng-ping, CHE Wang-jun, CHENG Ya-ling. et al. Gender-dependent Expression of ERβ in AEC Ⅱ of Fetal Mice Exposed to Arsenic and Estrogen Receptor Antagonist[J]. Journal of Sichuan University (Medical Sciences), 2018, 49(3): 364-368.
Citation: YANG Meng-ping, CHE Wang-jun, CHENG Ya-ling. et al. Gender-dependent Expression of ERβ in AEC Ⅱ of Fetal Mice Exposed to Arsenic and Estrogen Receptor Antagonist[J]. Journal of Sichuan University (Medical Sciences), 2018, 49(3): 364-368.

砷暴露及雌激素受体拮抗剂对雌、雄胎鼠AECⅡ内ERβ表达的影响

Gender-dependent Expression of ERβ in AEC Ⅱ of Fetal Mice Exposed to Arsenic and Estrogen Receptor Antagonist

  • 摘要: 目的 观察砷暴露及雌激素受体拮抗剂(ICI182,780)对雌、雄胎鼠肺泡Ⅱ型上皮细胞(AECⅡ)内雌激素受体β(ERβ)表达的影响。方法 孕19~20 d ICR小鼠剖腹取胎鼠,分离、纯化雌性组和雄性组胎鼠AECⅡ。四甲基亚唑蓝(MTT)法确定亚砷酸钠(NaAsO2)染毒AECⅡ细胞的剂量。将AECⅡ细胞分为以不同剂量(低、中、高)NaAsO2染毒的砷暴露组,培养24 h;另取AECⅡ细胞为联合暴露组,分别加入二甲基亚砜(DMSO)(溶剂对照组)、NaAsO2 5 μmol/L(单独砷染毒组)、NaAsO2(5 μmol/L)+ICI182,780(1×10-4 mol/L)(砷+拮抗剂组),培养24 h;均设空白对照组(不加任何试剂)。24 h后,砷暴露组行流式细胞术测凋亡率,砷暴露组、联合暴露组行实时荧光定量PCR法和Western blot检测胎鼠AECⅡ内ERβ mRNA和蛋白的表达水平。结果 AECⅡ纯度达(87.0±2.5)%。MTT法将0.5(低)、1.25(中)、5(高) μmol/L的浓度设置为染毒剂量。砷暴露组中:雌、雄中、高剂量组细胞凋亡率均高于空白对照组(P<0.05),而雌、雄各剂量组间比较差异无统计学意义。雌性中、高剂量组ERβ mRNA和蛋白表达水平高于空白对照组(P<0.05)和同剂量组雄性(P<0.05);雄性各剂量组ERβ mRNA和蛋白表达水平与空白对照组比较差异无统计学意义。联合暴露组中:雌性单独砷染毒组ERβ mRNA和蛋白表达水平高于砷+拮抗剂组(P<0.01)及同剂量组雄性(P<0.05),且高于空白对照组和溶剂对照组(P<0.05),雄性各组间比较差异无统计学意义;雌、雄其余各组间比较差异无统计学意义。结论 砷暴露可使雌性胎鼠AECⅡ内ERβ表达上调且高于雄性,此过程可被雌激素受体拮抗剂阻断。

     

    Abstract: Objective To determine the effects of arsenic and estrogen receptor antagonist (ICI182,780) on the expression of estrogen receptor beta (ERβ) in alveolar Ⅱ epithelial cells (AECⅡ) of female and male mice. Methods Nineteen or twenty day fetus mice were obtained through caesarean section of ICR mice. Purified AECⅡ cells were separated from the female and male fetus, respectively, and confirmed using immunofluorescence staining. The cells were exposed to sodium arsenite (NaAsO2) at a low, medium, or high dosage determined by MTT and cultured for 24 h. The NaAsO2 (5 μmol/L) exposed cells were compared with those treated (for 24 h) with dimethyl sulfoxide (DMSO) or ICI182,780 (1×10-4 mol/L). Apoptosis rates of the cells were measured by flow cytometry. Real-time fluorescence quantitative PCR method and Western blot technique were used to detect the expression ofERβmRNA and protein in AECⅡ. Results Purity of AECⅡ cells reached (87.0±2.5)%. NaAsO2 exposure was set at a concentration of 0.5 (low), 1.25 (medium), and 5 (high) μmol/L. The cells exposed to medium and high dosage of NaAsO2 had higher apoptosis rates than the blank controls (P<0.05), without sex differences. Female cells exposed to medium and high dosage of NaAsO2 had higher levels of expressions ofERβmRNA and protein than the blank controls (P<0.05) and male cells exposed to the same dosage of NaAsO2 (P<0.05). No significant differences were found in the expressions ofERβmRNA and protein between the exposed male cells and the blank controls. ICI182,780 lowered the expression levels ofERβmRNA and protein in the female exposed cells (P<0.01). Conclusion Arsenic exposure increases expressions of AECⅡ’s ERβ, more so in female cells than in male cells. This can be blocked by estrogen receptor antagonists.

     

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