邻苯二甲酸酯和双酚A联合暴露对大鼠睾丸间质细胞活性的影响

Effects of DEHP and BPA exposure in combination on cell activity in rat testis Leydig cells in vitro

目的研究环境内分泌污染物邻苯二甲酸二(2-乙基)己酯(DEHP)和双酚A(BPA)对原代大鼠睾丸间质细胞活性的影响。方法采用酶消化与贴壁法相结合提取大鼠睾丸间质细胞,分别暴露于终浓度0(对照)、1、10、100、1 000μmol/L的DEHP和(或)BPA溶液24 h。采用CCK8法检测细胞活性。结果与对照组比较,100、1 000μmol/L的DEHP暴露组和10、100、1 000μmol/L BPA暴露组及各浓度DEHP+BPA联合暴露组大鼠睾丸间质细胞的抑制率均较高,差异均有统计学意义(P<0.05);且随着DEHP和(或)BPA暴露浓度的升高,大鼠睾丸间质细胞的抑制率均呈上升趋势。与相同浓度DEHP+BPA联合暴露组比较,1、1 000μmol/L的DEHP暴露组大鼠睾丸间质细胞的抑制率较低,1 000μmol/L的BPA暴露组大鼠睾丸间质细胞的抑制率较高,差异均有统计学意义(P<0.05)。1μmol/L的DEHP与BPA联合暴露对细胞活性抑制表现为协同作用,1 000μmol/L的DEHP与BPA联合暴露表现为拮抗作用,10和100μmol/L的DEHP与BPA联合暴露对抑制细胞活性未表现出交互作用。结论 DEHP和BPA均能显著影响睾丸间质细胞活性,且以BPA作用更加明显;低剂量(1μmol/L)的DEHP和BPA联合作用为协同效应,高剂量(1 000μmol/L)的DEHP和BPA联合作用为拮抗效应。

Objective To study the combined effects of di-2-ethylhexylphthate（DEHP） and bisphenol A（BPA） on cell activity in rat Leydig cells.Methods After isolation by enzyme digestion combined with adherent method,rat Leydig cells were cultured for 48 h,then were treated with DEHP（1,10,100,1 000 μmol/L）,BPA（1,10,100,1 000 μmol/L） and DEHP＋BPA（1μmol/L＋1 μmol/L,10 μmol/L＋10 μmol/L,100 μmol/L＋100 μmol/L,1 000 μmol/L＋1 000 μmol/L）,0.1% DMSO as the control.After 24 h of treatment,cell viability was detected by colorimetric method with CCK8 kit.Results Compared with the control group,significantly declined cell viability was found in DEHP（100,1000 μmol/L） groups,BPA（10,100,1 000 μmol/L） groups and DEHP＋BPA groups at all doses（P0.05）;Suppression of cell viability at 1 and 1 000 μmol/L DEHP was less than that of groups with the same dose of DEHP ＋BPA,but 1 000 μmol/L BPA exceeded the same dose of DEHP ＋BPA（P〈 0.05）.In the combined exposure,the suppression of cell viability existed synergistic effect in 1 μmol/L DEHP ＋1 μmol/L BPA group,antagonism effect in 1 000 μmol/L DEHP ＋1 000 μmol/L BPA group.Conclusion DEHP and BPA can influence rat Leydig cell viability,especially BPA.Combined effect of DEHP and BPA may present synergistic effect in low concentration and antagonism effect in high concentration.