【摘要】 目的 研究车前子醇提物对多囊卵巢综合征（PCOS）大鼠卵巢颗粒细胞自噬的作用，并探讨其可能作用机制。方法 选取动情周期规律SD大鼠125只，取24只灌胃羧甲基纤维素钠（CMC-Na）设为对照组，其余101只灌胃来曲唑混悬液构建PCOS大鼠模型。建模成功的96只大鼠分为模型组（n=24）、3MA组（n=24）、低剂量组（n=24）和高剂量组（n=24）。对照组、模型组灌胃0.9%氯化钠溶液，低、高剂量组按10、20mg/（kg·d）灌胃车前子醇提物，3MA组按10mg/（kg·d）灌胃自噬阻断剂3MA,连续14天。酶联免疫吸附（ELISA）法检测大鼠血清促卵泡生成素（FSH）、黄体生成素（LH）、睾酮（T）、雌二醇（E₂）水平；苏木精-伊红（HE）染色法观察卵巢组织病变形态；流式细胞仪检测卵巢颗粒细胞自噬率；蛋白质印迹法（Western blot法）检测微管结合蛋白轻链3（LC3）、抑癌基因p53、磷酸化雷帕霉素靶蛋白（p-mTOR）、磷酸化腺苷酸活化蛋白激酶（p-AMPK）的表达情况。结果 与对照组比较，模型组大鼠血清LH、T和FSH水平均升高，E₂水平下降；卵巢组织病变最明显；颗粒细胞自噬率升高；LC3Ⅱ/LC3Ⅰ和p-AMPK蛋白表达增加，p53和p-mTOR蛋白表达降低（P<0.05）。与模型组比较，3MA组、低剂量组、高剂量组LH、T和FSH水平降低，E₂水平升高，卵巢组织病变减轻，颗粒细胞自噬率降低，LC3Ⅱ/LC3Ⅰ和p-AMPK蛋白表达降低，p53和p-mTOR蛋白表达升高（P<0.05）。各组与模型组比较的指标变化中，3MA组的指标变化高于低剂量组，低于高剂量组，差异有统计学意义（P<0.05）。结论 车前子醇提物可调节PCOS大鼠内分泌代谢，可能与调控p53/AMPK信号通路，抑制颗粒细胞自噬有关。更多还原

【Abstract】 Objective To study the effect of the ethanol extract of Plantago Seed on autophagy of ovarian granulosa cells in polycystic ovary syndrome（PCOS） rats, and to explore its possible mechanism. Methods 125 SD rats with regular estrous cycle were selected.24 rats were given intragastric carboxymethylcellulose sodium（CMC-Na） as the control group, and the remaining 101 rats were intragastrically administered with letrozole suspension to construct a PCOS rat model. The 96 rats successfully modeled were divided into model group（n=24）, 3 MA group（n=24）, low-dose group（n=24） and high-dose group（n=24）.The control group and model group were given normal saline intragastrically, the low and high dose groups were given 10, 20 mg/（kg·d） and the 3 MA group was given 10 mg/（kg·d） Gavage the autophagy blocker 3 MA for 30 consecutive days. Enzyme-linked immunosorbent assay（ELISA） method to detect rat serum follicle stimulating hormone（FSH）,luteinizing hormone（LH）, testosterone（T）, estradiol（E₂） levels. Hematoxylin-eosin（HE） staining Method was used to observe the morphology of ovarian tissue lesions.Flow cytometry to detect autophagy rate of ovarian granulosa cells; Western blot method was used to detect microtubule-associated protein 1 light chain 3（LC3）, tumor suppressor gene p53, and rapamycin phosphate and the expression of prime target protein（p-mTOR） and phosphorylated adenylate-activated protein kinase（p-AMPK）. Results Compared with the control group, the serum LH, T and FSH levels of the model group increased, and the E₂ level decreased. The ovarian tissue lesions were the most obvious. The autophagy rate of granulosa cells increased. The expression of LC3Ⅱ/LC3Ⅰ and p-AMPK protein increased, and p53 and the expression of p-mTOR protein decreased（P<0.05）. Compared with the model group, the levels of LH, T and FSH in the 3 MA group, low-dose group, and high-dose group decreased, and the E₂ level increased. The ovarian tissue lesions were alleviated, and the rate of granulosa cell autophagy decreased, and the expression of LC3Ⅱ/LC3Ⅰ and p-AMPK protein decreased., p53 and p-mTOR protein expression increased（P<0.05）. In the index changes of each group compared with the model group, the index changes of the 3 MA group were higher than that of the low-dose group and lower than that of the high-dose group, and the difference was statistically significant（P<0.05）.Conclusion The ethanol extract of Plantago Seed can regulate the endocrine metabolism of PCOS rats, which may be related to the regulation of p53/AMPK signaling pathway and the inhibition of granular cell autophagy.更多还原