菟丝子总黄酮对多囊卵巢综合征大鼠模型的影响

目的:探讨菟丝子总黄酮对脱氢表雄酮(dehydroepiandrosterone,DHEA)联合人绒毛膜促性腺激素(human chorionic gonadotropin,HCG)致大鼠多囊卵巢综合征(olycystic ovary syndrome,PCOS)模型的影响。方法:70只SD大鼠,10只正常组除外,其余大鼠上午颈背部注射DHEA 0. 06 mg·g-1,下午皮下注射1. 5 U HCG/只,连续21 d。造模第16天,阴道涂片,实施动情周期监测。挑选成模大鼠60只,分为模型组,达英-35组(0. 339 2 mg·kg-1),菟丝子总黄酮高、中、低剂量组(0. 2,0. 1,0. 05 g·kg-1),每组10只。分组当天按照组别分别进行给药,正常组和模型组给予等体积溶剂,连续给药21 d。末次给药2 h后,腹主动脉取血,测血清睾酮(testosterone,T),雌二醇(estrogen,E2),促黄体生成素(luteinizing hormone,LH)和卵泡刺激素(follicle stimulating hormone,FSH)的含量;脱颈处死大鼠,取每只大鼠同一部位的卵巢固定于10%甲醛溶液中,进行苏木素-伊红(HE)染色光镜观察卵巢的形态变化;取另一相同部位的卵巢,免疫组化法测量卵巢中雄激素受体(androgen receptor,AR)和凋亡相关蛋白B淋巴细胞瘤-2 (B-cell lymphoma-2,Bcl-2)和Bcl-2相关X蛋白(Bcl-2-associated X protein,Bax),观察相关蛋白在卵巢中的表达情况;取下丘脑和垂体,免疫组化法检测下丘脑中促性腺激素释放激素(gonadotropinreleasing hormone,GnRH),垂体促性腺激素释放激素受体(gonadorelin releasing hormone receptor,GnRHR)的表达情况。结果:PCOS大鼠模型复制成功。与模型组比较,各给药组可以显著降低PCOS大鼠血清T,E2和LH含量,显著升高血清FSH含量(P 0. 01);显著升高卵巢Bcl-2水平(P 0. 01),不同程度升高垂体GnRHR水平(P 0. 05,P 0. 01);不同程度降低卵巢AR和Bax水平(P 0. 05,P 0. 01),显著降低下丘脑中GnRH水平(P 0. 01)和显著改善PCOS大鼠卵巢组织的病理性改变。结论:PCOS大鼠模型复制成功。菟丝子总黄酮可能通过调节雌雄激素分泌、抑制卵巢凋亡蛋白的表达、影响下丘脑-垂体-卵巢轴途径发挥对PCOS模型大鼠的保护作用。

Effect of Dodder Total Flavone on Polycystic Ovary Syndrome Rat Models

Objective: To investigate the effect of dodder total flavone on polycystic ovary syndrome (PCOS) rat models induced by dehydroepiandrosterone (DHEA) combined with human chorionic gonadotropin (HCG). Method: Except the blank group, the remaining rats were injected with DHEA 0.06 mg·g^-1 in the morning on the nucha and 1.5 U HCG in the afternoon for 21 consecutive days. On the 16^th day after the modeling, the vaginal smear was performed to monitor the estrus cycle. Sixty rats with successful modeling were selected and divided into model group, dacin-35 group, and high, middle and low-dose dodder total flavonoids groups, with 10 rats in each group. On the day of grouping, drugs were given respectively to the drug treatment groups, and the blank group and the model group were given equal volume solvent. The drugs were given continuously for 21 days. Blood was collected from abdominal aorta 2 h later after the final administration, serum levels of testosterone (T), estrogen (E₂), luteinizing hormone (LH) and follicle stimulating hormone(FSH) were measured; rats were put to death, and the ovaries at the same part of each rat were fixed in 10% formalin solution. htoxylin eosin (HE) staining was performed, and the morphological changes in the ovaries were observed by light microscopy; the same part of the ovary was taken, and androgen receptor (AR), B-cell lymphoma-2(Bcl-2) and Bcl-2-associated X protein(Bax) were measured in the ovary by immunohistochemistry to observe the expressions of relevant proteins in the ovary; the hypothalamus and pituitary were taken, and the expressions of gonadotropin-releasing hormone (GnRH) in hypothalamus and gonadorelin releasing hormone receptor (GnRHR) in pituitary were detected by immunohistochemistry. Result: PCOS rat model was successfully replicated. Serum levels of T, E₂ and LH were significantly reduced, and FSH level of PCOS was significantly increased in each drug treatment group (P<0.01). The pituitary GnRHR level was increased to different degrees (P<0.05,P<0.01); AR and Bax levels of the ovary were decreased to different degrees (P<0.05,P<0.01); hypothalamus GnRH level was significantly decreased, and pathological changes in ovarian tissue of PCOS rats were obvious (P<0.01) in each drug treatment group. Conclusion: The PCOS rat model was successfully replicated. Total flavonoids of dodder may play a protective role in PCOS model rats by regulating the secretion of androgen, inhibiting the expression of ovarian apoptotic protein and impacting the hypothalamic-pituitary-ovary axis pathway.