G蛋白偶联受体30在烹调油烟诱导大鼠卵巢颗粒细胞毒性中的作用
Role of G protein-coupled receptor 30 in cooking oil fumes-induced ovarian cytotoxicity in rats
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摘要:
目的 分析G蛋白偶联受体30（GPR30）在烹调油烟（COFs）诱导大鼠卵巢颗粒细胞毒性中作用，为COFs预防控制提供基础数据。
方法 提取雌性Wistar大鼠的卵巢颗粒细胞，5、25、100、400、800、1 600 μg/mL COFs体外染毒24h后，MTT法检测细胞生存率，流式细胞术检测细胞凋亡，放射免疫检测雌激素（E2）、孕酮（P4）、卵泡刺激素（FSH）和黄体生成素（LH）水平，RT-PCR检测GPR30、表皮生长因子受体（EGFR）、信号传导与转录激活因子3（STAT3）和胞外信号调节激酶（ERK）mRNA表达水平。
结果 COFs染毒24 h后，100~1 600 μg/mL剂量组颗粒细胞生存率明显低于对照组（P < 0.001）；25~400 μg/mL剂量组颗粒细胞凋亡率明显高于对照组（P < 0.05），而100~400 μg/mL剂量组P4、E2激素水平均明显低于对照组（P < 0.01）。100~400 μg/mL剂量组GPR30 mRNA及400μg/mL剂量组EGFR mRNA均明显低于对照组（P < 0.05或P < 0.01），100~400μg/mL剂量组STAT3、ERK mRNA均明显高于对照组（P < 0.01）。
结论 一定剂量COFs可导致卵巢颗粒细胞凋亡率上升和P4、E2激素水平下降。COFs这些不利影响与STAT3和ERK mRNA的表达水平上升有关。GPR30可能参与了雌激素对卵巢颗粒细胞调节作用。
Abstract:Objective To analyze the role of G protein-coupled receptor 30 (GPR30) in the cytotoxicity to rat ovarian granulose cells induced by cooking oil fumes (COFs), and provide basic data for prevention and control of potential COFs-related negative health effects.
Methods Ovarian granulosa cells were extracted from female Wistar rats. After 24 hours of exposure to different doses of COFs (5, 25, 100, 400, 800, and 1 600 μg/mL), the cell survival rate was measured by MTT method, apoptosis by flow cytometry, the levels of estrogen (E2), progesterone (P4), follicle stimulating hormone (FSH), and luteinizing hormone (LH) by radioimmunoassay, and the expression levels of GPR30, epidermal growth factor receptor (EGFR), signal transducers and activators of transcription 3 (STAT3), and extracellular regulated protein kinase (ERK) mRNA by RT-PCR.
Results After 24 hours of COFs exposure, the survival rates of granulosa cells in the 100-1600μg/mL groups were significantly lower than that in the control group (P < 0.001); the apoptosis rates of granulosa cells in the 25-400 μg/mL groups were significantly higher than that in the control group (P < 0.05); the P4 and E2 hormone levels in the 100-400 μg/mL groups were significantly lower than those in the control group (P < 0.01). The expression levels of GPR30 mRNA in the 100-400μg/mL groups and the level of EGFR mRNA in the 400μg/mL group were significantly lower than those in the control group (P < 0.05 or P < 0.01); the expression levels of STAT3 and ERK mRNA in the 100-400μg/mL groups were significantly higher than those in the control group (P < 0.01).
Conclusion A certain dose of COFs can lead to an increase in the apoptosis rate and a decrease in the P4 and E2 hormone levels of ovarian granulosa cells. These adverse effects of COFs are associated with increased expression levels of STAT3 and ERK mRNA. GPR30 may be involved in the regulation of estrogen on ovarian granulosa cell growth and development.
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