优思悦对多囊卵巢综合征模型大鼠卵巢功能的影响及其机制研究

摘要 目的：探讨优思悦对硫酸脱氢表雄酮（dehydroepiandrosterone，DHEA）诱导的多囊卵巢综合征模型大鼠体内性激素：睾酮( testosterone，T)、雌二醇( estradiol，E2 )、促黄体生成素( luteinizing hormone，LH)及卵泡刺激素( follicle－stimulating hormone，FSH)及PI3K /AKT信号通路的的影响。方法：将60只雌性SD大鼠随机均分为3组，包括空白组、模型组及治疗组。空白组每日颈部皮下注射0.2 mL大豆油，其余各组每日颈部皮下注射DHEA 60 mg/kg+0.2 mL大豆油，持续注射35 d。造模第21 d时，每日上午制备大鼠阴道涂片，将其放置在显微镜下观察，根据细胞形态判定大鼠的动情周期，无动情周期规律视为造模成功。造模第36 d时，连续4 w对大鼠进行灌胃，每天一次。检测大鼠阴道分泌物的细胞形态、大鼠卵巢中卵泡发育的情况。放射免疫法测定大鼠血清中T、E2、LH、FSH的含量；RT-qPCR法检测大鼠卵巢组织中PI3K /AKT信号通路相关因子（IＲS-1、AKT-2、CSK-3β、GLUT-4 mRNA和PTEN mRNA）的表达变化。结果：空白组大鼠的动情周期有规律性，而模型组大鼠的动情周期失去规律性，且模型组大鼠卵巢中的卵泡呈囊状扩张，血清中T、LH水平明显升高（P<0.05），同时卵巢组织中的IRS-1、AKT-2、CSK-3β、GLUT-4 mRNA表达减少（P<0.05），PTEN mRNA表达增多（P<0.05）。与模型组比较，优思悦可改善PCOS模型大鼠动情周期，降低血清T、LH水平（P<0.05），改善卵泡发育，减少囊状扩张卵泡的形成，上调IRS-1、AKT-2、CSK-3β、GLUT-4 mRNA表达量，下调PTEN mRNA 表达量。结论：优思悦可有效治疗DHEA诱导的PCOS模型大鼠，与卵巢中的PI3K /AKT信号通路相关因子的基因表达相关。

Study on the Effect and Mechanism of Uxel on Ovarian Function in Rats with Polycystic Ovary Syndrome

ABSTRACT Objective: To investigate the effects of Yousiyue on sex hormones (T, E2, LH, FSH) and PI3K /AKT signaling pathways in rats with polycystic ovary syndrome induced by dehydroephedrone sulfate. Methods: 60 female SD ratswere randomly divided into three groups, including blank group, model group and treatment group. In the blank group, 0.2ml soybean oil was injected subcutaneously to the neck every day, while in the other groups, DHEA 60 mg /kg+ 0.2 mL soybean oil was injected subcutaneously to the neck every day for 35 days. On the 21st day of modeling, rat vaginal smears were prepared every morning and observed under a microscope. The estrous cycle of the rats was determined according to the cell morphology, and the absence of estrous cycle rule was deemed as the success of modeling. At the 36th day of modeling, the rats were given gavage once a day for 4 weeks. The cell morphology of vaginal secretions and follicular development in ovaries of rats were detected. The contents of T, E2, LH and FSH in rat serum were determined by radioimmunoassay. The expression of PI3K /AKT signaling pathway related factors (IRS-1, AkT-2, CSK-3, Glut-4 mRNA and PTEN mRNA) in the ovarian tissue of rats were detected by RT-QPCR. Results: Compared with the blank group, the rats in the model group lost the regular estrus cycle, the follicles in the ovary showed cystic expansion, the serum T and LH levels were significantly increased (P<0.05), and the expressions of IRS-1, AkT-2, CSK-3, glut-4 mRNA were decreased (P<0.05), and the expressions of PTEN mRNA were increased (P<0.05). In the blank rats, the estrus cycle was regular, while in the model rats, the follicles in the ovary of the model rats showed cystic expansion, and the serum T and LH levels were significantly increased (P<0.05). Meanwhile, the expressions of IRS-1, Akt-2, CSK-3, glut-4 mRNA in the ovary tissues were decreased (P<0.05), and the expressions of PTEN mRNA were increased (P<0.01). Compared with the model group, Yousiyue can improve the oestrogen cycle of PCOS model rats, reduce serum T and LH levels (P<0.05), improve follicular development, reduce the formation of cystic expansion follicles, up-regulate the expression of IRS-1, Akt-2, CSK-3, glut-4 mRNA, and down-regulate the expression of PTEN mRNA. Conclusion: Yousiyue can effectively treat DHEA-induced PCOS model rats, which is related to the gene expression of PI3K /AKT signaling pathway related factors in the ovary.