补骨膏对绝经后骨质疏松症大鼠的骨保护作用和对OPG-RANK-RANKL信号通路的影响*

作者：罗 娟1,2，曾朝辉1,3，袁尚锋2，朱莹莹2，罗 杰2，彭真灵2，龙厚任1

单位：1.湖南中医药大学，湖南 长沙 410208； 2.株洲市中医伤科医院，湖南 株洲 412007； 3.湖南中医药高等专科学校，湖南 株洲 412006

引用：引用：罗娟，曾朝辉，袁尚锋，朱莹莹，罗杰，彭真灵，龙厚任.补骨膏对绝经后骨质疏松症大鼠的骨保护作用和对OPG-RANK-RANKL信号通路的影响[J].中医药导报,2026,32(1):7-13.

DOI：10.13862/j.cn43-1446/r.2026.01.002

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摘要：

目的：探究补骨膏对绝经后骨质疏松症（PMOP）大鼠的骨保护作用及其对骨保护素（OPG）-核因子κB受体激活蛋白（RANK）-核因子κB受体激活蛋白配体（RANKL）信号通路的调控作用。方法：将36只大鼠随机分为假手术组（9只）和手术组（27只），手术组采用双侧卵巢切除建立PMOP大鼠模型。造模成功后，将24只PMOP大鼠随机分为模型组、戊酸雌二醇组、补骨膏低剂量组、补骨膏高剂量组，然后予相应药物灌胃8周。骨密度仪检测股骨近端骨密度；Micro-CT三维重建分析股骨微结构；苏木精-伊红（HE）染色观察股骨组织病理形态；酶联免疫吸附试验（ELISA）法检测血清中骨碱性磷酸酶（BALP）、骨钙素（BGP）、OPG水平，测定盒检测血清中磷、钙水平；蛋白质印迹法（Western blotting）检测股骨组织中OPG、RANK、RANKL蛋白表达水平；RT-qPCR法检测股骨组织肿瘤坏死因子-α（TNF-α） mRNA、干扰素-γ（IFN-γ） mRNA、精氨酸酶-1（Arg-1） mRNA、转化生长因子-β1（TGF-β1） mRNA、基质金属蛋白酶-9（MMP-9）mRNA、OPG mRNA、RANK mRNA、RANKL mRNA表达水平。结果：假手术组大鼠股骨结构连续完整，骨小梁数目较多，形态较厚，结构致密；模型组大鼠股骨近端骨密度明显降低；补骨膏低剂量组、补骨膏高剂量组和戊酸雌二醇组大鼠股骨近端骨小梁数量、骨组织形态结构均得到不同程度改善。模型组大鼠骨密度及血清中钙、BALP、BGP、OPG水平均低于假手术组（P＜0.01），血清磷水平高于假手术组（P＜0.01）；补骨膏低剂量组、补骨膏高剂量组及戊酸雌二醇组大鼠骨密度及血清中钙、BALP、BGP、OPG水平均高于模型组（P＜0.05或P＜0.01），血清磷低于模型组（P＜0.01）。模型组大鼠股骨组织OPG蛋白相对表达量低于假手术组（P＜0.01），RANK、RANKL蛋白相对表达量均高于假手术组（P＜0.01）；补骨膏低剂量组、补骨膏高剂量组及戊酸雌二醇组大鼠股骨组织中OPG蛋白相对表达量高于模型组（P＜0.05或（P＜0.01），RANK、RANKL蛋白相对表达量均低于模型组（P＜0.01）。模型组大鼠股骨组织TNF-α mRNA、IFN-γ mRNA、MMP-9 mRNA、RANK mRNA、RANKL mRNA对表达量均高于假手术组（P＜0.01），Arg-1 mRNA、TGF-β1 mRNA、OPG mRNA对表达量均低于假手术组（P＜0.01）；补骨膏高剂量组及戊酸雌二醇组大鼠股骨组织TNF-α mRNA、IFN-γ mRNA、MMP-9 mRNA、RANK mRNA、RANKL mRNA相对表达量均低于模型组（P＜0.01），Arg-1 mRNA、TGF-β1 mRNA、OPG mRNA相对表达量均高于模型组（P＜0.01）。结论：补骨膏可能通过调控OPG-RANK-RANKL信号通路，抑制免疫炎症反应，调节骨基质胶原合成与降解，从而维持骨代谢平衡，改善PMOP大鼠骨密度及骨微结构病理损伤。

关键词：绝经后骨质疏松症；补骨膏；OPG-RANK-RANKL信号通路；炎症反应；骨保护；大鼠

Abstract：Objective: To investigate the osteoprotective effects of Bugugao on rats with postmenopausal osteoporosis (PMOP) and its regulatory role in the osteoprotegerin (OPG)-receptor activator of nuclear factor-κB (RANK)-RANK ligand (RANKL) signaling pathway. Methods: A total of 36 rats were randomly assigned to a sham-operated group (n=9) and an ovariectomized group (n=27). PMOP models were established by bilateral ovariectomy. After successful modeling, 24 PMOP rats were randomly divided into a model group, an estradiol valerate group, a low-dose Bugugao group, and a high-dose Bugugao group, followed by 8 weeks of intragastric administration of corresponding treatments. Bone mineral density (BMD) of the proximal femur was measured using a bone densitometer; femoral microstructure was analyzed by Micro-CT three-dimensional reconstruction; histopathological morphology of the femur was observed via hematoxylin-eosin (HE) staining; serum levels of bone alkaline phosphatase (BALP), osteocalcin (BGP), and OPG were detected by enzyme-linked immunosorbent assay (ELISA), while serum phosphorus and calcium levels were measured using assay kits; protein expression levels of OPG, RANK, and RANKL in femoral tissue were determined by Western blotting; expression levels of tumor necrosis factor-α (TNF-α) mRNA, interferon-γ (IFN-γ)  mRNA, arginase-1 (Arg-1) mRNA, transforming growth factor-β1 (TGF-β1) mRNA, matrix metalloproteinase-9 (MMP-9) mRNA, OPG mRNA, RANK mRNA, and RANKL mRNA in femoral tissue were detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Results: The sham-operated group exhibited continuous and intact femoral structure with numerous, thick, and densely arranged trabeculae; the model group showed significantly reduced BMD in the proximal femur; the low-dose Bugugao group, high-dose Bugugao group, and estradiol valerate group all displayed varying degrees of improvement in trabecular number and bone histomorphology in the proximal femur. Compared with the sham-operated group, the model group showed lower BMD and serum levels of calcium, BALP, BGP, and OPG (P<0.01), but higher serum phosphorus levels (P<0.01). Compared with the model group, the low-dose Bugugao group, high-dose Bugugao group, and estradiol valerate group showed higher BMD and serum levels of calcium, BALP, BGP, and OPG (P<0.05 or P<0.01), and lower serum phosphorus levels (P<0.01). The model group showed lower relative protein expression of OPG in femoral tissue than sham-operated group (P<0.01), while higher expression levels of RANK and RANKL proteins than sham-operated group (P<0.01). Compared with the model group, the expression of OPG protein was increased in the low-dose Bugugao group, high-dose Bugugao group, and estradiol valerate group (P<0.05 or P<0.01), while the expression of RANK and RANKL proteins was decreased in the low-dose Bugugao group, high-dose Bugugao group, and estradiol valerate group (P<0.01). The expression levels of TNF-α mRNA, IFN-γ mRNA, MMP-9 mRNA, RANK mRNA, and RANKL mRNA in the model group were higher than that of the sham-operated group (P<0.01), whereas the expression levels of Arg-1 mRNA, TGF-β1 mRNA, and OPG mRNA were lower than that of the sham-operated group (P<0.01). In the high-dose Bugugao group and estradiol valerate group, the expression of TNF-α mRNA, IFN-γ mRNA, MMP-9 mRNA, RANK mRNA, and RANKL mRNA was lower (P<0.01), while the expression of Arg-1 mRNA, TGF-β1 mRNA, and OPG mRNA was higher (P<0.01) compared with the model group. Conclusion: Bugugao may maintain bone metabolic balance and improve BMD and pathological damage to bone microstructure in PMOP rats by regulating the OPG-RANK-RANKL signaling pathway, inhibiting immune-inflammatory responses, and modulating the synthesis and degradation of bone matrix collagen.

Key words：postmenopausal osteoporosis; Bugugao; OPG-RANK-RANKL signaling pathway; inflammatory response; osteoprotective; rats

发布时间：2026-01-30

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