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| 补肾健脾方介导肠道稳态改善去卵巢大鼠的骨量丢失 |
| The Efficacy of a kidney-tonifying and spleen-strengthening prescription in mediating gastrointestinal homeostasis and mitigating bone loss in ovariectomized rats |
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| DOI：10.3969/j.issn.1006-7108.2024.12.001 |
| 中文关键词: 绝经后骨质疏松症 补肾健脾方 骨量丢失 肠道稳态 |
| 英文关键词:postmenopausal osteoporosis kidney-tonifying and spleen-strengthening prescription bone loss gastrointestinal homeostasis |
| 基金项目:广东省自然科学基金面上项目（2021A1515011463，2021A1515011247）；广州市科技计划（202201011761） |
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| 中文摘要: |
| 目的 探讨肠道稳态在去卵巢（OVX）大鼠骨量丢失中的机制及补肾健脾方的防治作用。方法 18只4月龄的雌性未孕SD大鼠随机平均分为假手术（SHAM）组、去卵巢（OVX）组、补肾健脾方（BSJP）组，每组6只。造模5 d后，SHAM组及OVX组予生理盐水灌胃，BSJP组予补肾健脾方灌胃，连续灌胃12周后取材。大鼠股骨的皮质骨及松质骨采用Micro-CT进行分析，小肠及股骨组织HE染色后镜下观察形态，TRAP染色后对破骨细胞数量进行统计，Western Blot检测肠道屏障关键蛋白Cldn3、小肠及骨组织内的免疫细胞关键标志物TNF-α、IL-17A及趋化因子CCL20、CXCR3、骨组织内破骨标志蛋白C-FOS、DC-STAMP、CTSK表达情况。结果 与OVX 组比较，BSJP组肠壁增厚，小肠绒毛结构破坏减轻，股骨内破骨活动减少，Micro-CT 骨微结构扫描提示BMD均明显升高；肠道屏障标志蛋白Cldn3蛋白表达上升，免疫细胞关键标志物TNF-α、IL17A及趋化因子CCL20、CXCR3蛋白表达均下降，破骨标志蛋白C-FOS、DC-STAMP、CTSK表达均下降。结论 去卵巢（OVX）大鼠发生骨质疏松的病理基础是肠道稳态失衡，补肾健脾方介导肠道稳态改善去卵巢（OVX）大鼠的骨量丢失。 |
| 英文摘要: |
| Objective To explore the mechanism of intestinal homeostasis in bone loss in ovariectomized (OVX) rats and the preventive and therapeutic effects of kidney tonifying and spleen strengthening formula.Methods A cohort of eighteen 4-month-old female Sprague-Dawley (SD) rats, which were nulliparous, were randomly assigned into three groups: a sham-operated control group (SHAM), an ovariectomized group (OVX), and a group treated with the kidney- tonifying and spleen-strengthening prescription (BSJP), with each group comprising six subjects. Following a 5-day post-operative recovery period, the SHAM and OVX groups were administered saline via gavage, whereas the BSJP group received the prescribed formulation via the same route for a duration of 12 weeks prior to sample collection. Analyses included Micro-CT imaging of the femur to evaluate bone density and microarchitecture, histological examination with hematoxylin and eosin (HE) staining of the small intestine and femur tissues, enumeration of osteoclasts through tartrate-resistant acid phosphatase (TRAP) staining, and Western Blot analyses to quantify the expression levels of the intestinal barrier protein Claudin-3 (Cldn3), pro-inflammatory cytokines TNF-α and IL17A, chemokines CCL20 and CXCR3, and osteoclast-associated proteins C-FOS, DC-STAMP, and Cathepsin K (CTSK) in both the intestinal and bone tissues. Results Comparative analysis between the OVX and BSJP groups revealed a significant thickening of the intestinal wall, attenuation of damage to the villous architecture of the small intestine, and a reduction in osteoclastic activity within the femur in the BSJP group. Micro-CT scans demonstrated a notable increment in bone mineral density (BMD) . Elevated expression of the intestinal barrier protein Cldn3 was observed in conjunction with a decrease in the levels of pro-inflammatory markers TNF-α, IL17A and chemokines CCL20, CXCR3 as well as a reduction in osteoclast-associated proteins C-FOS, DC-STAMP, and CTSK. Conclusion Osteoporosis in ovariectomized (OVX) rats stems from a dysregulation of gastrointestinal homeostasis. The administration of the kidney-tonifying and spleen-strengthening formula appears to restore this balance, thereby ameliorating the bone loss associated with OVX. |
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