双酚 A 诱发卵巢储备功能下降大鼠模型的 制备及其中医证候属性.

Objective To investigate the optimal preparation method of bisphenol A (BPA) -induced diminished ovarian reserve (DOR) rat models, and to clarify the attributes of traditional Chinese medicine (TCM) syndromes. Methods Eighteen clean female SD rats aged 3 and 6 months with regular estrous cycle were selected and were randomly divided into the low-dose, medium-dose and high-dose BPA groups. The rats in the low-dose, medium-dose and high-dose BPA groups were given 50，100，and 200 mg/kg BPA mixed suspension orally, once a day until they showed unregular estrous cycles. The general performance and vaginal smear were observed everyday. The estrous cycle of rats was observed by vaginal smear; the abdominal aortic blood of rats was collected, and serum was separated. Serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), anti-Müllerian hormone (AMH), and inhibin-B (INHB) were detected by ELISA. We took bilateral ovaries, observed the morphology of ovarian tissues, and calculated the proportion of follicles at all levels and the ovarian index. We chose the model preparation method used in the group which had the most significant change of the indexes as the best method. The 18 successful model rats were randomly divided into the control group, Yougui pill group, and Yangjingzhongyu decoction group, with 6 in each. The model rats in the control group， Yougui pill group and Yangjingzhongyu decoction group were given distilled water, Yougui pill for treatment of kidny-yang deficiency, and Yangjingzhongyu decoction for kidny-yin deficiency by 1 mL/100 g body weight, for 2 consecutive weeks， respectively. The changes of estrous cycle, symptoms of kidney-yang deficiency, ovarian tissue morphology, ovarian index, proportion of follicles at all levels, serum hormone, INHB and AMH levels were compared between these two groups. Results All model rats in the BPA groups aged 3 and 6 months had reduced activity and liked to crouch and bunch. The disturbance time of estrous cycles of model rats aged 3 month in the medium-dose BPA group was significant shorter as compared with that of the other groups, and they showed decreased levels of AMH and primordial follicles and increased atretic follicles. Therefore, the preparation method in the 3-month-old rat model of the medium-dose BPA group was the best method. One rat in the Yangjingzhongyu decoction group and 5 rats in Yougui pill group resumed regular estrous cycle while all rats in the control group continued to be in disorder. The number of regular estrus cycle recovery in the Yougui pill group （5）was higher than that in the control group（0）and Yangjingzhongyu decoction group (1), while the symptom of kidneyyang deficiency improved more significantly. The ovaries of rats in the control group were pale, small and hard, while those of the Yougui pill group were bigger and ruddy. The ovaries of Yougui pill group had good elasticity and multiple growth follicles on the surface. There were fewer follicles on the ovarian surface and poor elasticity in the rats of Yangjingzhongyu decoction group. Compared with the other two groups, the Yougui pill group had higher proportion of primordium follicle, lower atresia follicle, and higher AMH level (all P<0. 05). Conclusions The DOR animal models can be successfully prepared by intragastric administration of 100 mg/kg BPA in 3-month clean SD rats. The attribute of TCM syndrome of the rat models is kidney-yang deficiency by means of syndrome differentiation through formula. [ABSTRACT FROM AUTHOR]