[Effects of electroacupuncture on the secretion function of ovarian cells and kisspeptin/kiss1r system in rats with polycystic ovarian syndrome].

Xue-Dan Zhao, Zhi-Hao Li, Jun-Wei Hu, Yue-Lai Chen, Ge Xu
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Abstract

Objective: To observe the effects of electroacupuncture (EA) on hormone secretion function of ovarian granulosa cells and theca cells, as well as the expression changes of kisspeptin and kiss1r in rats with polycystic ovarian syndrome (PCOS), so as to explore the mechanism of EA for relieving ovarian dysfunction in PCOS rats.

Methods: Forty-eight SD female rats were randomly divided into control group, model group, EA group and flutamide group, with 12 rats in each group. PCOS rat model was replicated with the gavage of letrozole (0.1 mg/mL, 10 mL•kg-1•d-1). In the EA group, EA (2 Hz, 2 mA) was used to stimulate "Guanyuan" (CV4) for 20 min each time. In the flutamide group, flutamide solution (50 mg•kg-1•d-1) was administrated by gavage. The treatments were given once daily for 14 days in each group. After the modeling and treatment, the body and ovarian weights of the rats were measured, and the ovarian index was calculated. Using HE staining, the morphological changes of ovary were observed. ELISA was adopted to detect the contents of testosterone (T), luteinizing hormone (LH) and estradiol (E2) in serum, the contents of E2 and T in the culture medium of ovarian granulosa cells and theca cells, as well as the content of kisspeptin in the ovarian tissue. The positive expression of kisspeptin in ovary was observed by immunohistochemical method, and the protein expression of its receptor kiss1r was detected by Western blot.

Results: Compared with the control group, the body and ovarian weights, ovarian index, the contents of T and LH in serum and that of T in the culture medium of theca cells, as well as the content and positive expression of kisspeptin in ovary were all increased (P<0.01, P<0.05); and the content of E2 in the culture medium of granulosa cells was decreased (P<0.01) in the model group. When compared with the model group, in the EA and flutamide groups, the body and ovarian weights, ovarian index, the contents of T and LH in serum and that of T in the culture medium of theca cells, as well as the content and expression of kisspeptin in ovary were all decreased (P<0.01, P<0.05); and the content of E2 in the culture medium of granulosa cells was increased (P<0.05, P<0.01).

Conclusion: EA regulates the serum sex hormone levels, the secretion function of the ovarian granulosa cells and theca cells, and the ovarian kisspeptin/kiss1r protein expression in PCOS rats, showing the similar effect as receptor blockade intervention. It is suggested that the improvement of EA in ovarian dysfunction of PCOS rats may be related to the kisspeptin/kiss1r system.

[电针对多囊卵巢综合征大鼠卵巢细胞分泌功能及kisspeptin/kiss1r系统的影响]。
目的观察电针(EA)对多囊卵巢综合征大鼠卵巢颗粒细胞和卵巢癌细胞激素分泌功能的影响EA)对多囊卵巢综合征(PCOS)大鼠卵巢颗粒细胞和卵巢癌细胞激素分泌功能的影响,以及kisspeptin和kiss1r的表达变化,从而探讨EA缓解PCOS大鼠卵巢功能障碍的机制:将48只SD雌性大鼠随机分为对照组、模型组、EA组和氟他胺组,每组12只。复制 PCOS 大鼠模型,灌胃来曲唑(0.1 mg/mL,10 mL-kg-1-d-1)。在EA组中,使用EA(2赫兹,2毫安)刺激 "关元"。刺激 "关元"(CV4),每次20分钟。每次 20 分钟。在氟他胺组中,氟他胺溶液(50 毫克-千克-1-d-1)灌胃给药。每组每天给药一次,连续 14 天。建模和治疗后,测量大鼠体重和卵巢重量,并计算卵巢指数。用 HE 染色法观察卵巢的形态变化。采用酶联免疫吸附法检测睾酮(T)、促黄体生成素(LH)和雌二醇(E2)的含量。和雌二醇(E2)血清中黄体生成素(LH)和雌二醇(E2)的含量,卵巢颗粒细胞和卵巢细胞培养液中E2和T的含量,以及卵巢组织中kisspeptin的含量。免疫组化法观察卵巢中kisspeptin的阳性表达,Western blot检测其受体kiss1r的蛋白表达:结果:与对照组相比,EA组小鼠的体重、卵巢指数、血清中T和LH的含量、theca细胞培养液中T的含量、卵巢中kisspeptin的含量和阳性表达均升高(颗粒细胞培养液中PP2含量降低)(颗粒细胞培养液中PPP2含量升高)(PPC结论:EA能调节小鼠血清性激素水平:EA可调节PCOS大鼠血清性激素水平、卵巢颗粒细胞和theca细胞的分泌功能以及卵巢kisspeptin/kiss1r蛋白的表达,其作用与受体阻断干预相似。这表明EA对多囊卵巢综合征大鼠卵巢功能障碍的改善可能与kisspeptin/kiss1r系统有关。
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