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目的探讨己烯雌酚(DES)、邻苯二甲酸二丁酯(DBP)、丙基硫脲嘧啶(PTU)对小鼠垂体瘤神经细胞(GT1-7神经细胞)的增殖效应及其联合作用。方法 DES的剂量为0.1、1、10、100、1 000μmol/L,DBP的剂量为0.01、1、10、100、1 000μmol/L,PTU的剂量为0.000 1、0.01、1、100、1 000μmol/L。联合作用的剂量设计为DES0.1~1 000μmol/L,DBP 0.01μmol/L,PTU 0.000 1μmol/L。采用CCK-8法检测3种物质单独作用和联合作用24 h后GT1-7神经细胞存活率。结果 DES、DBP单独作用于细胞,随DES或DBP浓度的增大细胞存活率呈现先增加后降低的趋势。PTU单独作用于细胞,各组的细胞存活率与对照组比较有所增加,但基本维持在一定水平。DES、DBP、PTU单独作用时,其浓度分别低于10、100、1 000μmol/L时,GT1-7神经细胞存活率在90%以上。DES与DBP联合暴露时,细胞存活率随DES浓度的增加而降低。DES与PTU联合暴露时,细胞存活率随DES浓度增加表现为先增加后降低,DBP与PTU联合暴露时,细胞存活率随DBP浓度增加表现为先增加后降低。与DES单独作用体系比较,0.01μmol/L DBP和0.000 1μmol/L PTU同时加入到0~10μmol/L DES后细胞存活率明显增加,差异有统计学意义(P<0.05),而DES浓度为100μmol/L时细胞存活率由80%左右降至4%左右,差异有统计学意义(P<0.05)。结论 DES、DBP、PTU对GT1-7神经细胞的增殖效应呈先促进后抑制的趋势,3种物质在一定剂量下共同作用对GT1-7神经细胞增殖呈现联合作用。
Objective To investigate the proliferative effects of diethylstilbestrol (DES), dibutyl phthalate (DBP) and propylthiouracil (PTU) on mouse pituitary adenoma cells (GT1-7 neurons) and their combined effects. Methods The dose of DES was 0.1, 1, 10, 100 and 1 000 μmol / L, the dose of DBP was 0.01, 1, 10, 100 and 1 000 μmol / L, the dose of PTU was 0.000 1, / L. The combined dose of DES was 0.1-1 000 μmol / L, DBP 0.01 μmol / L and PTU 0.000 1 μmol / L. CCK-8 method was used to test the survival rate of GT1-7 nerve cells after 24 h treatment of the three substances alone and in combination. Results The results showed that the cell viability decreased with the increase of DES or DBP concentration. PTU alone in the cells, the cell survival rate of each group compared with the control group increased, but remained at a certain level. When the concentrations of DES, DBP and PTU alone were lower than 10, 100 and 1 000 μmol / L respectively, the survival rate of GT1-7 nerve cells was over 90%. When DES and DBP were combined, the cell survival rate decreased with the increase of DES concentration. When combined with PTU, the survival rate of cells increased first and then decreased with the increase of DES concentration. When DBP and PTU were exposed, the cell survival rate increased at first and then decreased with the increase of DBP concentration. Compared with DES alone system, the cell viability was significantly increased after 0.01μmol / L DBP and 0.000 1μmol / L PTU were simultaneously added to 0 ~ 10μmol / L DES, the difference was statistically significant (P <0.05), while the DES concentration was 100μmol / L cell survival rate decreased from about 80% to about 4%, the difference was statistically significant (P <0.05). Conclusions The proliferative effects of DES, DBP and PTU on the neurons of GT1-7 cells first increase and then decrease. The synergistic effect of these three substances on the proliferation of GT1-7 neurons appears to be synergistic.