目的 观察二十五味鬼臼丸对睾酮缺乏引起的大鼠骨质疏松症的防治作用。方法 摘除双侧睾丸构建骨质疏松症动物模型，3月龄雄性SD大鼠30只，随机分为假手术组、模型组及二十五味鬼臼丸低、中、高剂量（200、400、800 mg/kg）组。术后1周开始每天ig给药，二十五味鬼臼丸200、400、800 mg/kg溶于3 mL生理盐水配制等体积混悬液，二十五味鬼臼丸低、中、高剂量组分别ig 200、400、800 mg/kg二十五味鬼臼丸，模型组和假手术组ig等量生理盐水，相同时间点连续给药8周，末次给药后取材。HE染色观察各组大鼠股骨和腓肠肌组织病理变化；ELISA法检测血清骨钙素（BGP）、骨保护素（OPG）、骨碱性磷酸酶（BALP）、抗酒石酸酸性磷酸酶（TRAP）、Ⅰ型胶原C端肽β特殊序列（β-CTX-Ⅰ）和钙离子（Ca^2＋）含量；Micro-CT检测股骨骨微结构；RT-PCR法检测β-连环蛋白（β-catenin）和矮小相关转录因子2（Runx2）的mRNA表达；Western blotting法检测β-catenin和Runx2的蛋白表达。结果 与模型组比较，二十五味鬼臼丸中剂量组大鼠骨小梁紊乱和腓肠肌组织病理变化明显改善；血清BGP、BALP、OPG和Ca^2＋水平升高（P＜0.05、0.01），TRAP、β-CTX-Ⅰ水平降低（P＜0.01）；BMD、Tb.N升高（P＜0.01），Tb.Sp降低（P＜0.05）；β-catenin和Runx2 mRNA表达上调（P＜0.05、0.01），Runx2蛋白表达上调（P＜0.01）。结论 二十五味鬼臼丸具有防治睾酮缺乏引起的大鼠骨质疏松症作用，其作用机制可能与激活Wnt/β-catenin信号通路有关。

Objective To observe the preventive and therapeutic effects of the Ershiwuwei Guijiu Pills on osteoporosis induced by testosterone deficiency in rats. Methods The animal model of osteoporosis was established by removing bilateral testicles. Thirty male SD rats of 3 months old were randomly divided into sham operation group, model group and low, medium and high dose (200,400,800 mg/kg) groups of Ershiwuwei Guijiu Pills. One week after operation, the rats were given intragastric administration every day. Ershiwuwei Guijiu Pills 200, 400 and 800 mg/kg were dissolved in 3 mL normal saline to prepare an equal volume suspension. Ershiwuwei Guijiu Pills low, medium and high dose groups were ig administered with Ershiwuwei Guijiu Pills 200, 400 and 800 mg/kg, respectively. The model group and sham operation group were ig administered with the same amount of normal saline. At the same time point, the rats were given continuous administration for 8 weeks, and the samples were taken after the last administration. HE staining was used to observe the pathological changes in the femur and gastrocnemius muscle tissues. Serum levels of osteocalcin (BGP), osteoprotegerin (OPG), bone alkaline phosphatase (BALP), tartrate-resistant acid phosphatase (TRAP), type I collagen C-terminal peptide (β-CTX-I), and calcium ions (Ca²⁺) were measured by ELISA. The bone microstructure was assessed using Micro-CT, and RT-PCR was used to detect the mRNA expression of β-catenin and runt-related transcription factor 2 (Runx2). Western blotting was used to measure the protein expression of β-catenin and Runx2. Results Compared with the model group, the medium-dose group treated with Ershiwuwei Guijiu Pills showed significant improvements in trabecular bone disorder and gastrocnemius muscle tissue pathology. Serum levels of BGP, BALP, OPG, and Ca²⁺ were increased (P < 0.05, 0.01), while TRAP and β-CTX-I levels were decreased (P < 0.01). BMD and Tb.N were increased (P < 0.01), while Tb.Sp were decreased (P < 0.05). mRNA expression of β-catenin and Runx2 was upregulated (P < 0.05, 0.01), and protein expression of Runx2 was upregulated (P < 0.01). Conclusion Ershiwuwei Guijiu Pills has a preventive and therapeutic effect on osteoporosis induced by testosterone deficiency in rats, and its mechanism may be related to the activation of the Wnt/β-catenin signaling pathway.

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青海省科技厅项目（2023-ZJ-783）；青海大学青年科研基金项目（2023-QYY-1）；青海大学医学部中青年科研基金项目（2023-kyy）