CAJ | 학술논문

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、250 nmol/L).MEHP(10、50、250 nmol/L)和溶剂对照 (DMSO),作用细胞24 h.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(P450sec)mRNA水平;用Real time-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,250 nmol/L MEHP抑制StAR基因及P450scc基因表达(P均<0.05);250 nmol/L DEHP对CYP19基因表达有促进作用(P<0.05);而10、50 nmol/L DEHP及50、250 nmol/L MEHP对CYP19基因表达均有抑制作用(P<0.05). 除10 nmol/L DEHP外,DEHP及MEHP其它浓度组均能抑制PPARα、PPARβ基因的表达(P<0.05);10 nmol/L DEHP及MEHP对PPARγ基因表达有促进作用(P<0.05),而250 nmol/L MEHP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),10 nmol/L DEHP、50及250 nmol/L MEHP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.
목적:연구린분이갑산이(2-을기)기지(DEHP)급기활성중간산물린분이갑산단(2-을기)기지(MEHP)대란소과립세포급기격소합성、분비공능적영향. 방법:체외배양건강초단유ICR소서적란소과립세포,분별가입DEHP(10、50、250 nmol/L).MEHP(10、50、250 nmol/L)화용제대조 (DMSO),작용세포24 h.용RT-PCR법측정원대소서과립세포중류고순격소합성급성조절단백(StAR)화P450측련렬해매(P450sec)mRNA수평;용Real time-PCR법측정원대소서과립세포중방향화매(CYP19)화과양화물매체증식제격활수체(PPARα、PPARβ、PPARγ)mRNA수평;용매련면역법(EIA)검측란소과립세포상청액중자이순화잉동적수평. 결과:여대조조비교,250 nmol/L MEHP억제StAR기인급P450scc기인표체(P균<0.05);250 nmol/L DEHP대CYP19기인표체유촉진작용(P<0.05);이10、50 nmol/L DEHP급50、250 nmol/L MEHP대CYP19기인표체균유억제작용(P<0.05). 제10 nmol/L DEHP외,DEHP급MEHP기타농도조균능억제PPARα、PPARβ기인적표체(P<0.05);10 nmol/L DEHP급MEHP대PPARγ기인표체유촉진작용(P<0.05),이250 nmol/L MEHP대PPARγ기인표체유억제작용(P<0.05).각제량DEHP급MEHP균능사과립세포분비자이순증가(P<0.05),10 nmol/L DEHP、50급250 nmol/L MEHP억제과립세포분비잉동(P<0.05). 결론:DEHP급MEHP영향과립세포류고순격소합성매、PPARs적기인표체급분비공능.
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, PPARγ were determined by Real time RT-PCR. Estradiol and progesterone were measured by enzyme immunoassay (EIA) in ovarian granulosa cell. RESULTS: 250 nmol/L MEHP group decreased the gene expression of StAR and P450scc (P<0.05); 250 nmol/L DEHP group increased the gene expression of CYP19, but 10, 50 nmol/L DEHP and 50, 250 nmol/L MEHP groups decreased the gene expression of CYP19 (P<0.05). Except 10 nmol/L DEHP group, DEHP and MEHP decreased the gene expression of PPARα and PPARβ (P<0.05). 10 nmol/L DEHP and MEHP groups increased the gene expression of PPARα, but 250 nmol/L MEHP group decreased the gene expression of PPARγ (P<0.05). 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 (P<0.05). CONCLUSION: DEHP and MEHP could affect the gene expression of steroid biosynthesis enzymes, PPARs, and the secretory function in ovarian granulosa cells.