目的 觀察二十五味鬼臼丸對睪酮缺乏引起的大鼠骨質(zhì)疏松癥的防治作用。方法 摘除雙側(cè)睪丸構(gòu)建骨質(zhì)疏松癥動物模型，3月齡雄性SD大鼠30只，隨機(jī)分為假手術(shù)組、模型組及二十五味鬼臼丸低、中、高劑量（200、400、800 mg/kg）組。術(shù)后1周開始每天ig給藥，二十五味鬼臼丸200、400、800 mg/kg溶于3 mL生理鹽水配制等體積混懸液，二十五味鬼臼丸低、中、高劑量組分別ig 200、400、800 mg/kg二十五味鬼臼丸，模型組和假手術(shù)組ig等量生理鹽水，相同時間點連續(xù)給藥8周，末次給藥后取材。HE染色觀察各組大鼠股骨和腓腸肌組織病理變化；ELISA法檢測血清骨鈣素（BGP）、骨保護(hù)素（OPG）、骨堿性磷酸酶（BALP）、抗酒石酸酸性磷酸酶（TRAP）、Ⅰ型膠原C端肽β特殊序列（β-CTX-Ⅰ）和鈣離子（Ca^2＋）含量；Micro-CT檢測股骨骨微結(jié)構(gòu)；RT-PCR法檢測β-連環(huán)蛋白（β-catenin）和矮小相關(guān)轉(zhuǎn)錄因子2（Runx2）的mRNA表達(dá)；Western blotting法檢測β-catenin和Runx2的蛋白表達(dá)。結(jié)果 與模型組比較，二十五味鬼臼丸中劑量組大鼠骨小梁紊亂和腓腸肌組織病理變化明顯改善；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表達(dá)上調(diào)（P＜0.05、0.01），Runx2蛋白表達(dá)上調(diào)（P＜0.01）。結(jié)論 二十五味鬼臼丸具有防治睪酮缺乏引起的大鼠骨質(zhì)疏松癥作用，其作用機(jī)制可能與激活Wnt/β-catenin信號通路有關(guān)。

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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青海省科技廳項目（2023-ZJ-783）；青海大學(xué)青年科研基金項目（2023-QYY-1）；青海大學(xué)醫(yī)學(xué)部中青年科研基金項目（2023-kyy）