毒理学杂志

目的 开展塑化剂邻苯二甲酸二环己酯(dicyclohexyl phthalate,DCHP)对人卵巢颗粒细胞KGN生成性腺类固醇激素干扰效应的研究，探讨其作用机制。方法 采用CCK-8的方法检测染毒剂量0.01～100μmol/L的DCHP对KGN细胞活性的影响;采用化学发光法检测DCHP无细胞毒性染毒剂量对KGN细胞生成雌二醇、睾酮和孕酮的改变;利用实时荧光定量PCR的方法检测3种激素相关基因表达量的变化。结果 DCHP可对KGN细胞产生细胞毒性，但是无细胞毒性染毒组培养液中雌二醇和孕酮的浓度明显降低，而睾酮的浓度明显增加(雌二醇组F=4.46、孕酮组F=10.07、睾酮组F=5.09,P<0.05);基因检测的结果显示KGN细胞中类固醇激素合成急性调节蛋白基因(star)(F=3.54,P<0.05)和芳香化酶P450基因(cyp19a1)(F=15.55,P<0.05)的表达量发生了明显下调，17β-羟基类固醇脱氢酶1(hsd17b1)(F=8.12,P<0.05)的表达量发生了明显上调。结论 DCHP主要通过改变star和cyp19a1基因的表达量干扰了KGN生成性腺类固醇激素，可能会导致生殖问题，因此需要关注DCHP对人生殖健康的潜在危害。

Objective A study was carried out to explore the interference effects of plasticizer dicyclohexyl phthalate( DCHP) on the synthesis of gonadal steroid hormones in human ovarian granulosa cells KGN and to extrapolate potential mechanism. Methods The influence of DCHP on the viability of KGN cells was detected by the CCK-8 method with the dose from 0. 01-100 μmol/L. The synthesis of estradiol,testosterone and progesterone in the culture medium of KGN cells after DCHP treatment were detected by the chemiluminescence method. The detection of changes in the expression of the main genes of the three hormones were performed by the reverse transcription realtime fluorescence quantitative PCR method. Results 100 μmol/L DCHP could cause cytotoxicity to KGN cells. The concentrations of estradiol and progesterone in the culture medium of the non cytotoxic group were significantly decreased( F = 4. 46,F = 10. 07,P < 0. 05),while the concentration of testosterone was significantly increased( F = 5. 09,P<0. 05). The expression levels of the steroid hormone synthesis acute regulatory protein gene( star) and estrogen aromatase cytochrome P450 gene( cyp19a1) were down regulated( F = 3. 54,F = 15. 55,P<0. 05),and the expressions of 17β-hydroxy steroid dehydrogenase 1( hsd17b1) was up regulated( F = 8. 12,P<0. 05). Conclusion DCHP could affect the synthesis of gonadal steroid hormones in KGN cells by modifying the expression levels of star and cyp19a1,which might cause potential reproductive harm. So the risk of DCHP to reproductive health need to be concerned.