Qinfeng Zhou, Kaixuan Wang, Cong Wang, Xiaoxian Sun, Lining Wang, Jie Sun, Yalan Pan, Muzhe Li, Zitong Zhao, Shijie Zhou, Qing Wang, Yafeng Zhang, Yong Ma, Yang Guo
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However, the specific mechanisms by which WSTLZTD exerts its anti-SOP effects-particularly through modulating macrophage senescence-remain unclear.</p><p><strong>Objective: </strong>The study aims to elucidate the role of WSTLZTD in macrophage senescence and SOP.</p><p><strong>Materials and methods: </strong>Aged mice received low, medium, high-dose WSTLZTD. Bone loss was evaluated via micro-computed tomography, hematoxylin and eosin staining and osteocalcin, tartrate-resistant acid phosphatase marker analysis. Macrophage senescence detection (β-galactosidase staining, p16, p21) and molecular mechanisms by Western blot, immunohistochemistry, immunofluorescence method were investigated. Macrophage-conditioned medium's effects on BMSC osteogenesis and mitochondrial function were assessed through alkaline phosphatase, Alizarin Red S staining, reactive oxygen species and JC-1 mitochondrial membrane potential (ΔΨm) assays.</p><p><strong>Results: </strong><i>In vivo</i> experiments demonstrated that WSTLZTD effectively ameliorated macrophage senescence and osteoporosis in naturally aged mice. Mechanistically, high-dose WSTLZTD attenuated senescence in bone marrow-derived macrophages by mediating LONP1, concurrently suppressing the cyclic GMP-AMP synthase (cGAS)/STING signaling pathway in BMSCs, thereby enhancing osteogenic differentiation of BMSCs. <i>In vitro</i> studies further confirmed that WSTLZTD-containing serum attenuated the senescent phenotype of macrophages. Notably, the LONP1 inhibitor, LONP1-IN-2, was found to diminish the anti-senescence effects of WSTLZTD on macrophages and BMSC osteogenesis.</p><p><strong>Discussion and conclusion: </strong>WSTLZTD potentially modulate macrophage senescence via LONP1, which subsequently suppresses the activation of the cGAS/STING pathway in BMSCs, ultimately promoting their osteogenic differentiation and ameliorating osteoporosis.</p>","PeriodicalId":19942,"journal":{"name":"Pharmaceutical Biology","volume":"63 1","pages":"524-548"},"PeriodicalIF":4.8000,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12305870/pdf/","citationCount":"0","resultStr":"{\"title\":\"Wen-Shen-Tong-Luo-Zhi-Tong Decoction alleviates bone loss in aged mice by suppressing LONP1-mediated macrophage senescence.\",\"authors\":\"Qinfeng Zhou, Kaixuan Wang, Cong Wang, Xiaoxian Sun, Lining Wang, Jie Sun, Yalan Pan, Muzhe Li, Zitong Zhao, Shijie Zhou, Qing Wang, Yafeng Zhang, Yong Ma, Yang Guo\",\"doi\":\"10.1080/13880209.2025.2537125\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Context: </strong>Aging leads to senile osteoporosis (SOP), marked by bone loss and increased fracture risk. 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引用次数: 0
摘要
背景：衰老导致老年性骨质疏松症（SOP），以骨质流失和骨折风险增加为特征。巨噬细胞作为骨组织中的活性免疫细胞，在衰老过程中对骨髓间充质干细胞（BMSCs）的成骨分化起着重要作用。温肾通络止痛汤（WSTLZTD）是一种治疗SOP的传统中药方剂，其疗效已被临床证实。然而，WSTLZTD发挥其抗sop作用的具体机制-特别是通过调节巨噬细胞衰老-尚不清楚。目的：研究WSTLZTD在巨噬细胞衰老和SOP中的作用。材料和方法：老龄小鼠低、中、高剂量WSTLZTD。通过显微计算机断层扫描、苏木精和伊红染色、骨钙素、抗酒石酸酸性磷酸酶标记分析评估骨质流失。采用Western blot、免疫组织化学、免疫荧光法检测巨噬细胞衰老（β-半乳糖苷酶染色、p16、p21）及其分子机制。通过碱性磷酸酶、茜素红S染色、活性氧和JC-1线粒体膜电位（ΔΨm）检测，评估巨噬细胞条件培养基对BMSC成骨和线粒体功能的影响。结果：体内实验表明，WSTLZTD能有效改善自然衰老小鼠巨噬细胞衰老和骨质疏松。机制上，大剂量WSTLZTD通过介导LONP1减轻骨髓源性巨噬细胞的衰老，同时抑制骨髓间充质干细胞中环GMP-AMP合成酶(cGAS)/STING信号通路，从而增强骨髓间充质干细胞的成骨分化。体外研究进一步证实，含wstlztd的血清可减轻巨噬细胞的衰老表型。值得注意的是，LONP1抑制剂LONP1- in -2被发现可以减弱WSTLZTD对巨噬细胞和BMSC成骨的抗衰老作用。讨论与结论：WSTLZTD可能通过LONP1调控巨噬细胞衰老，从而抑制骨髓间质干细胞中cGAS/STING通路的激活，最终促进其成骨分化，改善骨质疏松症。
Wen-Shen-Tong-Luo-Zhi-Tong Decoction alleviates bone loss in aged mice by suppressing LONP1-mediated macrophage senescence.
Context: Aging leads to senile osteoporosis (SOP), marked by bone loss and increased fracture risk. Macrophages, as active immune cells in bone tissue, play an important role in osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) during aging. Wen-Shen-Tong-Luo-Zhi-Tong Decoction (WSTLZTD), a traditional Chinese herbal formula, has been clinically validated for its efficacy in treating SOP. However, the specific mechanisms by which WSTLZTD exerts its anti-SOP effects-particularly through modulating macrophage senescence-remain unclear.
Objective: The study aims to elucidate the role of WSTLZTD in macrophage senescence and SOP.
Materials and methods: Aged mice received low, medium, high-dose WSTLZTD. Bone loss was evaluated via micro-computed tomography, hematoxylin and eosin staining and osteocalcin, tartrate-resistant acid phosphatase marker analysis. Macrophage senescence detection (β-galactosidase staining, p16, p21) and molecular mechanisms by Western blot, immunohistochemistry, immunofluorescence method were investigated. Macrophage-conditioned medium's effects on BMSC osteogenesis and mitochondrial function were assessed through alkaline phosphatase, Alizarin Red S staining, reactive oxygen species and JC-1 mitochondrial membrane potential (ΔΨm) assays.
Results: In vivo experiments demonstrated that WSTLZTD effectively ameliorated macrophage senescence and osteoporosis in naturally aged mice. Mechanistically, high-dose WSTLZTD attenuated senescence in bone marrow-derived macrophages by mediating LONP1, concurrently suppressing the cyclic GMP-AMP synthase (cGAS)/STING signaling pathway in BMSCs, thereby enhancing osteogenic differentiation of BMSCs. In vitro studies further confirmed that WSTLZTD-containing serum attenuated the senescent phenotype of macrophages. Notably, the LONP1 inhibitor, LONP1-IN-2, was found to diminish the anti-senescence effects of WSTLZTD on macrophages and BMSC osteogenesis.
Discussion and conclusion: WSTLZTD potentially modulate macrophage senescence via LONP1, which subsequently suppresses the activation of the cGAS/STING pathway in BMSCs, ultimately promoting their osteogenic differentiation and ameliorating osteoporosis.
期刊介绍：
Pharmaceutical Biology will publish manuscripts describing the discovery, methods for discovery, description, analysis characterization, and production/isolation (including sources and surveys) of biologically-active chemicals or other substances, drugs, pharmaceutical products, or preparations utilized in systems of traditional medicine.
Topics may generally encompass any facet of natural product research related to pharmaceutical biology. Papers dealing with agents or topics related to natural product drugs are also appropriate (e.g., semi-synthetic derivatives). Manuscripts will be published as reviews, perspectives, regular research articles, and short communications. The primary criteria for acceptance and publication are scientific rigor and potential to advance the field.
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