DEHP及MEHP对小鼠卵巢颗粒细胞分泌功能的影响

Effects of di(2-ethylhexyl) phthalate and mono(2-ethyl-hexyl) phthalate on ovarian granulosa cells in mice

目的:研究邻苯二甲酸二(2_乙基)己酯(DEHP)及其活性中间产物邻苯二甲酸单(2_乙基)己酯(MEHP)对卵巢颗粒细胞及其激素合成、分泌功能的影响。方法:体外培养健康初断乳ICR小鼠的卵巢颗粒细胞,分别加入DEHP(10、50、250nmol/L),MEHP(10、50、250nmol/L)和溶剂对照(DMSO),作用细胞24h。用RT-PCR法测定原代小鼠颗粒细胞中类固醇激素合成急性调节蛋白(StAR)和P450侧链裂解酶(P450scc)mRNA水平;用Realtime-PCR法测定原代小鼠颗粒细胞中芳香化酶(CYP19)和过氧化物酶体增殖剂激活受体(PPARα、PPARβ、PPARγ)mRNA水平;用酶联免疫法(EIA)检测卵巢颗粒细胞上清液中雌二醇和孕酮的水平。结果:与对照组比较,250nmol/LMEHP抑制StAR基因及P450scc基因表达(P均<0.05);250nmol/LDEHP对CYP19基因表达有促进作用(P<0.05);而10、50nmol/LDEHP及50、250nmol/LMEHP对CYP19基因表达均有抑制作用(P<0.05)。除10nmol/LDEHP外,DEHP及MEHP其它浓度组均能抑制PPARα、PPARβ基因的表达(P<0.05);10nmol/LDEHP及MEHP对PPARγ基因表达有促进作用(P<0.05),而250nmol/LMEHP对PPARγ基因表达有抑制作用(P<0.05)。各剂量DEHP及MEHP均能使颗粒细胞分泌雌二醇增加(P<0.05),10nmol/LDEHP、50及250nmol/LMEHP抑制颗粒细胞分泌孕酮(P<0.05)。结论:DEHP及MEHP影响颗粒细胞类固醇激素合成酶、PPARs的基因表达及分泌功能。

OBJECTIVE：To investigate the effects of di（2-ethylhexyl） phthalate （DEHP） and its active metabolite , mono（2-ethylhexyl） phthalate （ MEHP） on ovarian granulosa cells .METHODS ：ICR mice ages at the 21 days were used by subcutaneous injection PMSG. The ovarian granulosa cells were collected and cultured in DMEM-F12. After the ovarian granulosa cells were prepared, cells were exposed to doses of 10, 50, 250 nmol/L DEHP and 10, 50, 250 nmol/L MEHP for 24 h. The mRNA levels of StAR, P450scc were determined by RT-PCR. The mRNA levels of CYP19, PPARα, PPARβ, PPARγwere determined by Real ti me RT-PCR. Estradiol and progesterone were measured by enzyme i mmunoassay （EIA） in ovarian granulosa cell .RESULTS ：250 nmol/L MEHP group decreased the gene expression ofStARandP450scc（P0.05） ; 250 nmol/L DEHP group increased the gene expression ofCYP19, but 10, 50 nmol/L DEHPand 50, 250 nmol/L MEHP groups decreased the gene expression ofCYP19（P0.05） . Except 10 nmol/L DEHP group, DEHP and MEHP decreased the gene expression ofPPARαandPPARβ（P0.05） . 10 nmol/L DEHP and MEHP groups increased the gene expression ofPPARγ, but 250 nmol/L MEHP group decreased the gene expression ofPPARγ（P0.05） . DEHP and MEHP enhanced the concentration of estradiol at all dose groups , but 10 nmol/L DEHP, 50 and 250 nmol/L MEHP groups decreased the concentration of progesterone in ovarian granulosa cells （P0.05） .CONCLUSION：DEHP and MEHP could affect the gene expression of steroid biosynthesis enzymes , PPARs , and the secretory function in ovarian granulosa cells .