中国骨质疏松杂志

补肾化痰方调控自噬水平影响BMSCs骨/脂代谢平衡治疗PMOP研究

Effect of Bushen Huatan Decoction on autophagy level and bone / lipid metabolism balance of BMSCs in the treatment of PMOP

DOI：10.3969/j.issn.1006-7108.2026.07.004

中文关键词: 补肾化痰方绝经后骨质疏松自噬骨髓间充质干细胞

英文关键词:Bushen Huatan Formula postmenopausal osteoporosis autophagy bone marrow mesenchymal stem cells

基金项目:国家自然科学基金资助项目（82074416，82305350）；湖北省中医药创新发展联合基金项目（2022CFD153，2024AFD296）；第七批全国老中医药专家学术经验继承项目工作（国中医药人教函2022-76号）；湖北省卫生和计划生育委员会湖北中医大师名师评选暨工作室建设工作项目（鄂卫生计生办通[2018]32号）

作者	单位
陈杰1 周广文1 王育恒1 熊梦欣2 李都1 刘馥溧1 杨硕硕1 陈之浩1 向楠1*	1. 湖北中医药大学,湖北武汉 430060 2. 湖北中医药大学附属医院/湖北省中医院,湖北武汉 430074

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中文摘要:

目的探讨补肾化痰方含药血清对骨髓间充质干细胞（BMSCs）成骨/成脂分化及自噬活性的调控作用，阐明其防治绝经后骨质疏松症（PMOP）的潜在机制。方法采用制备补肾化痰方含药血清干预BMSCs；采用CCK-8法检测细胞活力并筛选最佳干预浓度；通过茜素红和油红O染色观察矿化结节及脂滴形成；运用Western blot检测成骨（BMP2、RUNX2、Collagen I）、成脂（PPARγ、C/EBPα、C/EBPβ）及自噬相关蛋白（Atg13、Beclin1、LC3Ⅱ/Ⅰ）表达；采用Real-time PCR检测自噬相关基因表达；透射电镜观察自噬体数量。结果补肾化痰方含药血清能增加矿化结节形成，减少脂滴生成；上调成骨分化标志物（BMP2、RUNX2、Collagen I）蛋白表达（P<0.05），下调成脂标志物（PPARγ、C/EBPα、C/EBPβ）表达（P<0.05）；提升自噬相关蛋白及基因（Atg13、Beclin1、LC3Ⅱ/Ⅰ）表达水平（P<0.05），并增加自噬体数量。结论补肾化痰方可能通过激活BMSCs自噬活性，促进成骨分化并抑制成脂分化，调节骨/脂代谢平衡，从而发挥防治PMOP作用。

英文摘要:

Objective This study aimed to explore the modulatory effects of Bushen Huatan prescription–containing serum on the osteogenic and adipogenic differentiation potential of bone marrow-derived mesenchymal stem cells (BMSCs), as well as its influence on autophagic activity, in order to elucidate its possible mechanism in preventing and managing postmenopausal osteoporosis (PMOP). Methods BMSCs were exposed to medicated serum prepared with the Bushen Huatan formula. Cell viability was assessed via the CCK-8 assay to identify the optimal concentration for intervention. The extent of extracellular matrix mineralization and intracellular lipid accumulation was evaluated using Alizarin Red S and Oil Red O staining, respectively. Western blotting was applied to examine protein levels associated with osteogenesis (BMP2, RUNX2, Collagen I), adipogenesis (PPARγ, C/EBPα, C/EBPβ), and autophagy (Atg13, Beclin1, LC3-II/I). Gene expression linked to autophagy was quantified through quantitative real-time polymerase chain reaction (qRT-PCR). The presence and abundance of autophagic structures were further confirmed by transmission electron microscopy (TEM). Results Serum containing the Bushen Huatan prescription significantly enhanced matrix mineral deposition and reduced lipid storage in BMSCs. The expression of osteogenic markers (BMP2, RUNX2, Collagen I) was markedly elevated (P < 0.05), while levels of adipogenic markers (PPARγ, C/EBPα, C/EBPβ) were suppressed (P < 0.05). Moreover, both transcriptional and translational indicators of autophagy-including Atg13, Beclin1, and the LC3-II/I ratio-showed significant upregulation (P < 0.05). TEM analysis revealed an increased formation of autophagosomes following treatment. Conclusion Bushen Huatan–derived serum may exert its protective effects against PMOP by enhancing autophagic flux in BMSCs, thereby promoting osteoblastic differentiation and inhibiting adipogenic commitment. This dual regulation contributes to restoring the bone–fat homeostatic balance in the bone marrow microenvironment.

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