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| 补肾健脾活血方改善去势小鼠骨微结构与肌肉质量 |
| Bushen Jianpi Huoxue Formula ameliorates bone microstructure and muscle mass in ovariectomized mice |
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| DOI：10.3969/j.issn.1006-7108.2026.06.002 |
| 中文关键词: 肌少-骨质疏松症 去势小鼠 补肾健脾活血方 骨密度 肌肉质量 |
| 英文关键词:sarcopenia-osteoporosis ovariectomized mice Bushen Jianpi Huoxue Formula bone mineral density (BMD) muscle mass |
| 基金项目:国家自然科学基金(82274551,82174395)；广东省科技计划项目（社会发展科技协同创新体系建设）-与衰老相关的骨质疏松症防治关键技术及特色中草药优选研究（2024A1111120018）；广东省自然科学基金面上项目（2022A1515012067） |
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| 中文摘要: |
| 目的 研究补肾健脾活血方对卵巢切除小鼠骨骼及肌肉功能的影响。方法 30只9周龄SPF级雌性C57BL/6J小鼠，采用随机数字表分为假手术组（n=10）和模型组（n=20），模型组实施双侧卵巢切除，假手术组仅切除卵巢周围相应体积的脂肪组织，将模型组小鼠按随机数字表分为模型对照组（OVX组）、补肾健脾活血方组（BSJPHXF组），每组10只，BSJPHXF组予以补肾健脾活血方（生药量为1.43 g/mL，剂量为0.017 mL/g），OVX组、假手术组接受相同体积的生理盐水，各组均采用灌胃方式给药，持续干预6周。监测小鼠体重，利用DXA检测小鼠全身骨密度、脂肪和瘦组织百分比；利用抓力测定仪检测小鼠造模前、给药3周及6周后四肢抓力；Micro-CT、HE染色分析股骨骨小梁微结构、测定小鼠腓肠肌湿重比、腓肠肌HE染色分析肌纤维横截面积。结果 DXA结果显示，与OVX组比较，BSJPHXF组全身骨密度、瘦组织含量增高，而脂肪组织百分比降低（P＜0.05）；Micro-CT分析提示，与OVX组比较，BSJPHXF组BV/TV、Tb.N升高，而Tb.Sp降低（P＜0.05）；股骨HE染色同样提示与OVX组相比，BSJPHXF组股骨骨小梁面积增多（P＜0.05）；与OVX组比较，BSJPHXF组OCN及RUNX2蛋白表达上升（P＜0.05）；四肢抓力结果提示在给药前，各组间小鼠四肢抓力差异不具有统计学意义（P＞0.05），给药3周及6周后，BSJPHXF组小鼠四肢抓力均高于OVX组（P＜0.05）；同时肌肉HE染色结果显示，BSJPHXF组小鼠腓肠肌横截面积大于OVX组（P＜0.05）。WB结果显示，与OVX组比较，BSJPHXF组MYF5和MYOD蛋白表达上升（P＜0.05）。结论 中药补肾健脾活血方具有提高去势小鼠骨密度、改善骨微结构及提高四肢肌力、增加瘦组织含量的作用。 |
| 英文摘要: |
| Objective To investigate the effects of the Bushen Jianpi Huoxue Formula (BSJPHXF) on bone and muscle function in ovariectomized mice. Methods Thirty 9-week-old female SPF C57BL/6J mice were randomly assigned to a sham surgery group (n=10) and a model group (n=20) using a random number table. The model group underwent bilateral ovariectomy (OVX), while the sham surgery group had an equivalent volume of peri-ovarian adipose tissue removed. The model group mice were then further divided randomly into an OVX control group (OVX group) and a BSJPHXF treatment group (BSJPHXF group), with 10 mice per group. The BSJPHXF group received BSJPHXF (crude drug concentration: 1.43 g/ml; dosage: 0.017 ml/g body weight). The OVX group and sham surgery group received an isovolumetric dose of physiological saline. All groups were administered the treatments via gavage for 6 weeks. Body weight was monitored. Dual-energy X-ray absorptiometry (DXA) was used to measure whole-body bone mineral density (BMD) and the percentage of whole-body fat tissue and lean tissue. A grip strength meter was used to measure limb grip strength before modeling, after 3 weeks of treatment, and before sacrifice. Micro-CT and hematoxylin and eosin (HE) staining were used to analyze femoral trabecular bone microstructure. The gastrocnemius muscle wet weight ratio was determined. HE staining of the gastrocnemius muscle was performed to analyze the cross-sectional area (CSA) of muscle fibers. Results DXA results showed that compared to the OVX group, the BSJPHXF group exhibited increased whole-body BMD and lean tissue content, but decreased fat tissue percentage (P< 0.05). Micro-CT analysis indicated that compared to the OVX group, the BSJPHXF group had increased BV/TV (bone volume fraction) and Tb.N (trabecular number), and decreased Tb.Sp (trabecular separation) (P< 0.05). Femoral HE staining similarly revealed increased trabecular bone area in the BSJPHXF group compared to the OVX group (P< 0.05). Compared to the OVX group, protein expression of OCN (osteocalcin) and RUNX2 was upregulated in the BSJPHXF group (P< 0.05). Limb grip strength results showed no statistically significant difference among groups before treatment (P> 0.05). After 3 weeks (3w) and 6 weeks (6w) of treatment, limb grip strength in the BSJPHXF group was significantly higher than in the OVX group (P< 0.05). Concurrently, muscle HE staining showed that the gastrocnemius muscle fiber CSA was larger in the BSJPHXF group than in the OVX group (P< 0.05). Western blotting (WB) results showed that protein expression of MYF5 and MYOD was increased in the BSJPHXF group compared to the OVX group (P< 0.05). Conclusion The Chinese herbal formula BSJPHXF improves bone mineral density, ameliorates bone microstructure, increases limb muscle strength, and enhances lean tissue content in ovariectomized mice. |
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