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目的:验证一种多囊卵巢综合征(PCOS)动物模型的建模效果,并针对模型研究黄体生成素受体(LHR)、胰岛素受体(INSR)和雄激素(AR)基因DNA甲基化状态。方法:选取23日龄SD雌性大鼠,模型组每日皮下注射脱氢表雄酮(DHEA)0.6mg/kg,对照组每日皮下注射等体积溶剂。注射20d后观察大鼠卵巢形态学(HE染色)、性激素(E2、T、P、FSH、LH)及空腹血糖和胰岛素的变化,并计算HOMA指数以评价胰岛素抵抗状况。建模成功后利用甲基化特异性PCR技术(MSP)检测模型组和对照组大鼠LHR、INSR和AR基因的DNA甲基化状态。结果:模型组大鼠的体质量显著低于对照组(P=0.000);去除体质量的影响后,模型组大鼠卵巢的质量略重于对照组,但无显著性差异(P=0.317)。模型组卵巢各级发育期卵泡及黄体少见,卵泡多呈囊性扩张,卵泡膜细胞、间质细胞增生;血清E2、T、FSH、FINS、FBG均显著高于对照组(均P<0.05);LH水平较对照组有所升高,但无显著性差异;LH/FSH比值组间无显著性差异,HOMA指数显著高于对照组(P=0.000)。LHR和AR基因经甲基化特异性模型组均未检出甲基化条带;INSR基因经MSP检测出甲基化条带,甲基化率为73.3%,显著高于对照组(16.7%)(P=0.000)。结论:利用DHEA成功诱导SD幼年雌性大鼠出现与PCOS患者相似的卵巢病理改变及雄激素升高和胰岛素抵抗等典型内分泌变化,是一种较为理想的PCOS动物模型。胰岛素抵抗、高胰岛素血症可能是肾上腺源性雄激素诱发PCOS的重要原因。DNA甲基化介导的胰岛素受体基因转录抑制是对PCOS胰岛素抵抗发生机制的一种全新补充。
OBJECTIVE: To validate the modeling effect of an animal model of polycystic ovary syndrome (PCOS) and to study the DNA methylation status of LHR, INSR and AR genes. . Methods: 23-day-old SD female rats were selected. DHEA 0.6 mg / kg was injected subcutaneously in the model group, and the control group was injected subcutaneously with an equal volume of solvent. Morphological (HE staining), sex hormones (E2, T, P, FSH, LH) and fasting blood glucose and insulin in rats were observed 20 days after injection, and HOMA index was calculated to evaluate the status of insulin resistance. After modeling, methylation-specific PCR (MSP) was used to detect the DNA methylation status of LHR, INSR and AR genes in the model group and the control group. Results: The body weight of rats in model group was significantly lower than that in control group (P = 0.000). After removal of body weight, the quality of ovary in model group was slightly higher than that of control group, but no significant difference (P = 0.317) . The ovarian follicles and corpus luteum at the developmental stage were rare in the model group, the follicles were mostly cystic dilatation, and the cells of the follicular cells and interstitial cells proliferated. The levels of E2, T, FSH, FINS and FBG in the model group were significantly higher than those in the control group (all P <0.05) ; LH levels increased compared with the control group, but no significant difference; LH / FSH ratio group no significant difference, HOMA index was significantly higher than the control group (P = 0.000). The methylation bands of methylation of INSR gene were detected by MSP. The methylation rate of INSR gene was 73.3%, which was significantly higher than that of control group (16.7% ) (P = 0.000). Conclusions: The ovarian pathological changes similar to PCOS patients and the typical endocrine changes such as androgen and insulin resistance appear to be induced by DHEA in SD young female rats, which is an ideal animal model of PCOS. Insulin resistance, hyperinsulinemia may be an important reason for adrenogenic androgen induced PCOS. DNA methylation-mediated insulin receptor gene transcription inhibition is a new complement to the mechanism of PCOS insulin resistance.