邻苯二甲酸-单-乙基己基酯对人卵巢颗粒细胞活性及分泌功能的影响

目的研究邻苯二甲酸-单-乙基己基酯[Mono-(2-ethylhexyl)phthalate,MEHP]对人卵巢黄素化颗粒细胞的活性及分泌功能的影响。方法选取北京大学人民医院行体外受精-胚胎移植患者19例,收集其卵泡液,提取颗粒细胞进行原代细胞培养72h,并分为低剂量组(10例,MEHP25μmol/L、50μmol/L、100μmol/L)和高剂量组(9例,MEHP300μmol/L、500μmol/L),两组均于加药后48h,采用细胞计数试剂盒检测颗粒细胞的活性、放免法检测雌二醇及孕酮水平,均与DMEM/F12培养液+二甲基亚砜(对照组)比较。结果高剂量组中,当MEHP≥500μmol/L时,颗粒细胞活性较对照组明显降低(P=0.001);当MEHP≥500μmol/L时,雌二醇水平较对照组明显降低(P=0.000);低剂量组中,当MEHP为50μmol/L、100μmol/L时,孕酮水平较对照组明显降低(P分别为0.007和0.02);高剂量组中,当MEHP为500μmol/L时,孕酮水平明显降低(P=0.002)。结论高浓度MHEP(≥500μmol/L)不仅降低人卵巢颗粒细胞活性,而且抑制了雌孕激素合成的功能。但目前人体暴露范围内(≤50μmol/L)的MHEP对颗粒细胞活性及雌激素合成无影响,对孕激素合成有明显抑制作用。

The effect of mono-(2-ethylhexyl)phthalate on human granulosa cells viability and steroid production

Objective To investigate the effect of Mono-（2-ethylhexyl）phthalate（MEHP）on viability and steroid production of human granulosa cells.Methods Samples of follicular fluid were obtained from nineteen patients who had undergone in vitro fertilization at Peking University People＇s Hospital.Human granulosa cells were extracted and cultured in vitro for 72 hours.Ten cases（low-dose group）were cultured in medium with low concentrations of MEHP（25,50,100 μmol/L）.Nine cases（high-dose group）were cultured with MEHP of 300,500 μmol/L.After 48 hours,granulosa cells viability was determined.Progesterone and estradiol were determined with radioimmuno-scintigraphy.The results were compared with negative control group（DMEM/F12 medium＋ DMSO）.Results The viability of granulosa cells was decreased obviously when the concentration of MEHP was ≥500 μmol/L（P=0.001）.The estradiol level was reduced significantly in the presence of MEHP ≥500 μmol/L（P=0.000）.In the low-dose group,progesterone levels were reduced significantly with 50 μmol/L and 100 μmol/L MEHP（P=0.007,0.02）,meanwhile,in the high-dose group,MEHP of ≥500 μmol/L reduced the progesterone level（P=0.002）.Conclusions MHEP reduced the activity of（≥500 μmol/L）human ovarian granulosa cells,inhibited synthesis of estrogen and progesterone.The current human exposure（≤50 μmol/L）of MHEP had no effect on the activity of granulosa cells and estradiol synthesis,while progesterone production was inhibited significantly.