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目的研究邻苯二甲酸二丁酯(dibutyl phthalate,DBP)对大鼠卵巢颗粒细胞中细胞凋亡因子Bcl-2、Bax表达的影响。方法体外培养25 d雌性SD大鼠卵巢颗粒细胞,分别加入DBP使终浓度分别(0、5、20、80μmol/L),培养24 h。采用放射免疫方法测定雌二醇(estradiol,E2)和孕酮(progestone,P)的水平;采用实时荧光定量PCR(real-time PCR)和免疫印迹试验(Western blot)检测卵巢颗粒细胞中Bcl-2、Bax mRNA和蛋白的表达。结果 DBP染毒后颗粒细胞分泌的E2及P水平下降,各剂量DBP均可降低颗粒细胞中Bcl-2 mRNA和蛋白的表达(P<0.05),且表达量与DBP剂量呈反比,同时,DBP可以增加颗粒细胞中Bax mRNA和蛋白的表达(P<0.05),且表达量与DBP剂量呈正比。结论 DBP可导致大鼠卵巢颗粒细胞凋亡,且Bcl-2/Bax比例失调可能是导致其凋亡的机制。
Objective To investigate the effect of dibutyl phthalate (DBP) on the expression of Bcl-2 and Bax in rat ovarian granulosa cells. Methods Ovarian granulosa cells of 25-day-old female Sprague-Dawley rats were cultured in vitro. The final concentration of DBP was added to the ovarian granulosa cells (0, 5, 20 and 80μmol / L) respectively and cultured for 24 hours. The levels of estradiol (E2) and progestone (P) were determined by radioimmunoassay. Real-time PCR and Western blot were used to detect the expression of Bcl- 2, Bax mRNA and protein expression. Results The levels of E2 and P secreted by granulosa cells after DBP exposure were decreased. DBP at various dosages decreased the expression of Bcl-2 mRNA and protein in granulosa cells (P <0.05), and the expression levels of DBP Can increase the expression of Bax mRNA and protein in granulosa cells (P <0.05), and the expression level is proportional to the dose of DBP. Conclusion DBP can induce the apoptosis of ovarian granulosa cells in rats, and the imbalance of Bcl-2 / Bax may be the mechanism of apoptosis.