目的 基于下丘脑-垂体-卵巢轴（hypothalamus-pituitary-ovary axis，HPOA）探讨藏药二十五味鬼臼丸对去卵巢大鼠骨质疏松的影响。方法 将40只SD雌性未孕大鼠随机分为假手术组、模型组、雌二醇组、二十五味鬼臼丸组，每组10只。除假手术组仅摘取卵巢旁同样大小的脂肪块，其余3组大鼠均摘除双侧卵巢建立绝经后骨质疏松症（postmenopausal osteoporosis，PMOP）大鼠模型。手术1周后开始给药处理，雌二醇组ig 0.1 mg/kg戊酸雌二醇，二十五味鬼臼丸组ig 441 mg/kg二十五味鬼臼丸，假手术组和模型组ig等体积蒸馏水，1次/d，连续12周后，ELISA法检测血清雌二醇、黄体生成素（luteinizing hormone，LH）、卵泡刺激素（follicle stimulating hormone，FSH）、促性腺激素释放激素（gonadotropin releasing hormone，GnRH）水平；RT-PCR检测下丘脑、垂体、左侧股骨远端雌激素受体α（estrogen receptor α，ERα）、雌激素受体β（estrogen receptor β，ERβ）mRNA表达；免疫荧光染色法检测下丘脑、垂体和右侧股骨远端ERα、ERβ蛋白表达；HE染色法观察大鼠右侧股骨远端组织形态变化；Micro-CT检测右侧股骨远端微结构的改变。结果 藏药二十五味鬼臼丸能显著降低大鼠体内血清LH、FSH、GnRH水平（P<0.05、0.01），并在HPOA轴中显著性上调下丘脑及垂体ERα mRNA及蛋白表达（P<0.01），同时显著性上调股骨ERβ mRNA及蛋白表达（P<0.01），并能显著回调大鼠股骨密度及骨微结构参数（P<0.01）。结论 藏药二十五味鬼臼丸可有效防治PMOP，其机制可能与药物直接作用骨组织雌激素受体或作用HPOA轴靶器官相关雌激素受体后，通过改变体内性激素的水平间接调控骨代谢有关。

Objective Based on the hypothalamus-pituitary-ovary axis (HPOA) to explore the effect of Tibetan medicine Ershiwuwei Guijiu Pill (二十五味鬼臼丸) on osteoporosis in ovariectomized rats. Methods Forty non-pregnant SD female rats were randomly divided into sham operation group, model group, estradiol group, and Ershiwuwei Guijiu Pill group, with 10 rats in each group. Except for the sham operation group, only the fat masses of the same size beside the ovaries were removed, and the rats in the other three groups were removed bilateral ovaries to establish a rat model of postmenopausal osteoporosis (PMOP). One week after the operation, the administration was started. The estradiol group was given 0.1 mg/kg estradiol valerate, the Ershiwuwei Guijiu Pill group was given 441 mg/kg Ershiwuwei Guijiu Pill. The same volume of distilled water were given to the sham operation group and the model group and gavage daily for 12 consecutive weeks. The levels of serum estradiol, luteinizing hormone (LH), follicle stimulating hormone (FSH), and gonadotropin releasing hormone (GnRH) were detected by ELISA; RT-PCR was used to detect estrogen receptor alpha (ERα) and estrogen receptor beta (ERβ) mRNA expression in the hypothalamus, pituitary, and the distal femur of the left side; Immunofluorescence staining method was used to detect the expression of ERα and ERβ protein in the hypothalamus, pituitary and distal right femur; HE staining method was used to observe the morphological changes of the distal femur of rats; Micro-CT was used to detect the microstructure changes of the distal femur of the right. Results The Tibetan medicine Ershiwuwei Guijiu Pill can significantly decrease serum LH, FSH, GnRH levels in rats (P < 0.05, 0.01), and significantly up-regulate ERα mRNA and protein expression of hypothalamus and pituitary in the HPOA axis (P < 0.01), and significantly up-regulate femoral ERβ mRNA and protein expression (P < 0.01). And it can significantly recall the femur bone density and bone microstructure parameters of rats (P < 0.01). Conclusion The Tibetan medicine Ershiwuwei Guijiu Pill can effectively prevent and treat PMOP, and its mechanism may be related to the direct action of the drug on the estrogen receptor in bone or the indirect regulation of bone by changing the level of sex hormones in the body after acting on the estrogen receptor subtypes on the target organs of the HPOA.

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青海省科技厅基础研究项目（2018-ZJ-730，2019-SF-134）；中央高校基本科研业务费专项资金资助（31920180028）