骨宝胶囊调控核因子κB信号通路抑制骨细胞凋亡促进成骨分化治疗骨质疏松的相关机制

doi:10.6040/j.issn.1671-7554.0.2026.0025

摘要/Abstract

摘要： 目的探讨骨宝胶囊调控核因子κB(nuclear factor-kappa B, NF-κB)信号通路抑制骨细胞凋亡促进成骨分化治疗骨质疏松的相关机制。方法通过网络药理学方法检索骨宝胶囊药物活性成分相关靶点、成骨分化和骨质疏松相关靶点,并获取三者交集。将交集靶点通过String数据库绘制蛋白质相互作用网络图并通过Cytoscape 3.8.0进行拓扑计算,选取degree值位于中位数以上的靶点作为核心靶点进行基因本体论(gene ontology, GO)与京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes, KEGG)富集分析。动物实验选取36只SD大鼠分为对照组(不做处理)、模型组(通过去势卵巢法构建骨质疏松模型)和用药组(在模型组基础上给予骨宝胶囊灌胃处理),每组12只。干预完成后通过苏木精-伊红染色、天狼猩红染色、甲苯胺蓝染色和番红固绿染色观察各组大鼠骨组织病理结构改变;通过原位末端标记法和免疫荧光法检测各组大鼠骨组织中细胞凋亡情况;通过全景病理学分析检测骨组织NF-κB信号通路激活情况;通过免疫组织化学染色法检测各组大鼠骨组织成骨相关蛋白骨形态发生蛋白 2、骨桥蛋白、成骨相关转录因子和Runt 相关转录因子 2表达。结果检索骨宝胶囊药物作用靶点643个,成骨分化相关靶点2 998个,骨质疏松相关靶点7 707个,三者交集点为266个。degree值位于中位数以上的核心交集靶点有132个。KEGG富集分析显示,核心交集靶点主要富集在成骨分化和NF-κB等信号通路;GO生物过程发现核心交集靶点主要细胞凋亡、程序性细胞凋亡等。动物实验发现,与对照组相比,模型组大鼠骨组织结构破坏严重,骨小梁断裂,纤维蛋白与蛋白聚糖流失。此外,伴随着NF-κB p65与Caspase3表达量的升高(P<0.05),模型组大鼠骨小梁中骨细胞凋亡率上升(P<0.05)。全景病理分析显示,模型组大鼠骨小梁骨组织中NF-κB信号通路存在异常激活,成骨相关蛋白表达量下降(P<0.05)。给予骨宝胶囊灌胃干预后,上述情况均出现明显逆转。结论骨宝胶囊可通过抑制骨小梁骨组织中NF-κB信号通路的异常激活,降低骨细胞凋亡率,上调成骨相关蛋白的表达,促进成骨分化从而发挥治疗骨质疏松的作用。

关键词: 骨宝胶囊, 核因子κB, 信号通路, 骨细胞凋亡, 成骨分化, 骨质疏松

Abstract: Objective To explore the mechanism by which Gubao Capsule treats osteoporosis by regulating the nuclear factor-kappa B(NF-κB)signaling pathway to inhibit osteocyte apoptosis and promote osteogenic differentiation. Methods Network pharmacology was used to retrieve targets related to the active ingredients of Gubao Capsule, osteogenic differentiation, and osteoporosis, and their overlapping targets were obtained. A protein-protein interaction(PPI)network was constructed using the STRING database, and topological analysis was performed with Cytoscape 3.8.0. Core targets(degree values above the median)were selected for gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analyses. In animal experiments, 36 SD rats were divided into three groups: control group(no treatment, n=12), model group(ovariectomy-induced osteoporosis, n=12), and treatment group(Gubao Capsule gavage based on the model group, n=12). After intervention, hematoxylin-eosin, Sirius red, toluidine blue, and safranin O-fast green staining were performed to observe bone pathological changes. TUNEL and immunofluorescence assays were used to detect osteocyte apoptosis. Panoramic pathological analysis evaluated NF-κB pathway activation. Immunohistochemistry detected the expression of osteogenic proteins: bone morphogenetic protein 2(BMP2), osteopontin(OPN), osterix, and runt-related transcription factor 2(RUNX2). Results A total of 643 targets of Gubao Capsule, 2,998 osteogenic differentiation-related targets, and 7,707 osteoporosis-related targets were retrieved, with 266 overlapping targets. Among them, 132 core targets with degree values above the median were identified. KEGG enrichment showed that the core targets were mainly enriched in osteogenic differentiation and NF-κB signaling pathways; GO analysis revealed that theenrichment in biological processes such as apoptosis and programmed cell death.Animal experiments showed that compared with the control group, the model group exhibited severe bone tissue destruction, trabecular fracture, and loss of fibrin and proteoglycans. Meanwhile, NF-κB p65 and Caspase3 expression were significantly upregulated(P<0.05), accompanied by a marked increase in osteocyte apoptosis rate(P<0.05). Panoramic pathological analysis confirmed abnormal activation of the NF-κB pathway and significantly decreased osteogenic protein expression in the model group(P<0.05). All these abnormalities were significantly reversed after Gubao Capsule intervention. Conclusion Gubao Capsule can treat osteoporosis by inhibiting abnormal activation of the NF-κB signaling pathway in trabecular bone tissue, reducing osteocyte apoptosis, upregulating osteogenic protein expression, and promoting osteogenic differentiation.

Key words: Gubao Capsule, Nuclear factor-kappa B, Signaling pathway, Osteocyte apoptosis, Osteogenic differentiation, Osteoporosis

中图分类号:

R274

闫小龙,陈东峰,管东辉,赵灿斌,邵将,王慧玺,罗峥伟,徐博洋,战颢志. 骨宝胶囊调控核因子κB信号通路抑制骨细胞凋亡促进成骨分化治疗骨质疏松的相关机制[J]. 山东大学学报 (医学版), 2026, 64(8): 35-47.

YAN Xiaolong, CHEN Dongfeng, GUAN Donghui, ZHAO Canbin, SHAO Jiang, WANG Huixi, LUO Zhengwei, XU Boyang, ZHAN Haozhi. Mechanism of Gubao Capsule in the treatment of osteoporosis by regulating the nuclear factor-κB signaling pathway to inhibit osteocyte apoptosis and promote osteogenic differentiation[J]. Journal of Shandong University (Health Sciences), 2026, 64(8): 35-47.

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