桃叶珊瑚苷通过抑制NF-κb信号通路减轻小鼠的膝骨关节炎

doi:10.12122/j.issn.1673-4254.2025.10.06

南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (10): 2104-2110.doi: 10.12122/j.issn.1673-4254.2025.10.06

• • 上一篇

桃叶珊瑚苷通过抑制NF-κb信号通路减轻小鼠的膝骨关节炎

麦泳欣¹^,²^,⁴(), 周舒婷²^,⁴, 温蕊嘉²^,⁴, 张锦芳¹(), 詹冬香²^,³^,⁴^,⁵()

^1.广州中医药大学深圳医院，广东深圳 518000
^2.广东省岭南特色医院制剂转化工程技术研究中心，广东广州 510405
^3.广东省中医临床研究院，广东广州 510405
^4.广州中医药大学第一附属医院，广东广州 510405
^5.医疗机构中药制剂与中药新药转化广东省工程研究中心，广东广州 510405

收稿日期:2025-03-14 出版日期:2025-10-20 发布日期:2025-10-24
通讯作者: 张锦芳,詹冬香 E-mail:20201110039@stu.gzucm.edu.cn;zhangjf06@gzucm.edu.cn;zdxbwjs@gzucm.edu.cn
作者简介:麦泳欣，博士，医师，E-mail: 20201110039@stu.gzucm.edu.cn
基金资助:
国家自然科学基金(82272526)

Aucubin alleviates knee osteoarthritis in mice by suppressing the NF‑κB signaling pathway

Yongxin MAI¹^,²^,⁴(), Shuting ZHOU²^,⁴, Ruijia WEN²^,⁴, Jinfang ZHANG¹(), Dongxiang ZHAN²^,³^,⁴^,⁵()

^1.Shenzhen Hospital (Futian) of Guangzhou University of Chinese Medicine, Shenzhen 518000, China
^2.Guangdong Engineering Technology Research Center of Commercialization of Lingnan Special Medical Institution Preparations, Guangzhou 510405, China
^3.Guangdong Provincial Academy of Clinical Research in Traditional Chinese Medicine, Guangzhou 510405, China
^4.First Hospital Affiliated to Guangzhou University of Chinese Medicine, Guangzhou 510405, China
^5.Guangdong Engineering Research Center of Commercialization of Medical Institution Preparations and Traditional Chinese Medicines, Guangzhou 510405, China

Received:2025-03-14 Online:2025-10-20 Published:2025-10-24
Contact: Jinfang ZHANG, Dongxiang ZHAN E-mail:20201110039@stu.gzucm.edu.cn;zhangjf06@gzucm.edu.cn;zdxbwjs@gzucm.edu.cn
Supported by:
National Natural Science Foundation of China(82272526)

摘要/Abstract

摘要：

目的探讨桃叶珊瑚苷通过调控NF-κB信号通路治疗膝骨关节炎（KOA）的分子机制。方法将60只SPF级C57BL/6J小鼠随机分为假手术组、模型组、氨基葡萄糖组（阳性对照）及桃叶珊瑚苷低、中、高剂量组（2、4、8 mg/kg），n=10。采用前交叉韧带切断术（ACL）建立KOA模型，术后第1天开始干预，1次/周，连续8周。通过番红固绿染色、免疫组化（IHC）检测软骨病理形态、炎症相关蛋白表达及μCT检测软骨下骨骨体积分数的变化情况（BV/TV）；Western blotting检测COL2、SOX9、p-P65、IL-1β及MMP13蛋白水平。构建软骨细胞KOA模型，采用阿利新蓝和番红O染色观察成软骨分化能力，RT-PCR检测COL2、SOX9及TNF-α基因表达。结果与模型组相比，桃叶珊瑚苷处理后COL2、SOX9蛋白表达增加，而pP65、IL-β1及MMP13蛋白表达降低（P<0.05）。桃叶珊瑚苷处理通过抑制NF-kb信号通路相关蛋白及MMP13的表达，促进COL2、SOX9等软骨相关蛋白的表达，增加软骨下骨骨体积分数。IL-1β诱导的软骨细胞模型中，RT-PCR结果显示，SOX9、COL2的基因表达在桃叶珊瑚苷组中表达增加，而TNF-α基因表达降低（P<0.05）。阿利新蓝和番红O染色结果显示，桃叶珊瑚苷促进软骨细胞外基质合成，增强成软骨分化能力。结论桃叶珊瑚苷通过抑制NF-κB信号通路减轻软骨炎症反应，促进软骨基质合成，改善软骨下骨微结构，从而有效治疗KOA。

关键词: 桃叶珊瑚苷, 骨关节炎, NF-κB, 软骨修复, 基质金属蛋白酶

Abstract:

Objective To assess the therapeutic effect of aucubin in mice with knee osteoarthritis (KOA) and investigate the underlying mechanism. Methods Sixty C57BL/6J mice were randomized equally into sham operation group, KOA model group, glucosamine (positive control) treatment group, and low-, medium-, and high-dose aucubin treatment groups (2, 4, and 8 mg/kg, respectively). KOA mouse models were established by transection of the anterior cruciate ligament (ACL), and the treatment was initiated on day 1 postoperatively and administered weekly for 8 weeks. Safranin O-fast green staining, immunohistochemistry, and microCT were used to evaluate the changes in cartilage pathology, inflammatory protein expression, and subchondral bone volume fraction (BV/TV). The expression levesl of COL2, SOX9, p-P65, IL-1β and MMP13 proteins in the cartilage tissues were detected using Western blotting. In a chondrocyte model with IL-1β treatment for mimicking KOA, the effect of aucubin on chondrogenic differentiation was observed with Alcian blue and Safranin O staining, and cellular COL2, SOX9 and TNF‑α mRNA expressions were detected with RT-qPCR. Results Compared with those in the model group, the mouse models receiving aucubin treatment showed significantly upregulated COL2 and SOX9 protein levels and downregulated p-P65, IL-1β and MMP13 expressions in the cartilage tissues. In the IL-1β-induced chondrocyte model, aucubin treatment significantly upregulated the mRNA expressions of SOX9 and COL2 but lowered the mRNA expression of TNF-α. Alcian blue and Safranin O staining confirmed that aucubin promoted the synthesis of cartilage extracellular matrix and enhanced chondrogenic differentiation of the cells. Conclusion Aucubin can effectively alleviate KOA in mice by inhibiting NF‑κB-mediated cartilage inflammation, promoting cartilage matrix synthesis, and improving subchondral bone microstructure.

Key words: aucubin, osteoarthritis, nuclear factor-κB, cartilage repair, matrix metalloproteinases

麦泳欣, 周舒婷, 温蕊嘉, 张锦芳, 詹冬香. 桃叶珊瑚苷通过抑制NF-κb信号通路减轻小鼠的膝骨关节炎[J]. 南方医科大学学报, 2025, 45(10): 2104-2110.

Yongxin MAI, Shuting ZHOU, Ruijia WEN, Jinfang ZHANG, Dongxiang ZHAN. Aucubin alleviates knee osteoarthritis in mice by suppressing the NF‑κB signaling pathway[J]. Journal of Southern Medical University, 2025, 45(10): 2104-2110.

图/表 6

表1 软骨基质合成及炎症反应相关引物序列

Tab.1 Primer sequences for RT-qPCR for amplifying mRNAs associated with cartilage matrix synthesis and inflammatory response

Gene	Primer sequences
COL2A1-Forward	GGCAATAGCAGGTTCACGTACA
COL2A1-Reverse	CGATAACAGTCTTGCCCCACTT
SOX9-Forward	TCCTCAGGCTTTGCGATTT
SOX9-Reverse	TGCTCGGGCACTTATTGG
TNF-α-Forward	CTGACGAATTACAGGGCCAAT
TNF-α-Reverse	TGCGTGAACCTCTTGAACAGT
18S-Forward	TGGTTGCAAAGCTGAAACTTAAAG
18S-Reverse	AGTCAAATTAAGCCGCAGGC

图1 膝关节软骨组织形态学分析

Fig.1 Morphological analysis of knee joint cartilage in mice. A: HE staining. B: Safranin-O/Fast Green staining.

图2 软骨下骨μCT三维重建及骨密度分析

Fig.2 MicroCT 3D reconstruction and bone mineral density analysis of the subchondral bone in the mouse models. *P<0.05, ***P<0.001 vs Model group.

图3 炎症相关蛋白免疫组化染色

Fig.3 Immunohistochemical staining for inflammatory proteins in the cartilage tissues of the mouse models. A: p65 staining. B: TNF-α staining. C, D: Quantitative analysis of p65 and TNF-α expression levels. *P<0.05, **P<0.01, ***P<0.001 vs Model group.

图4 蛋白表达水平的Western blotting分析

Fig.4 Western blotting of protein expressions in the chondrocyte model. A: Representative protein blots. B-F: Quantitative analysis of p-p65/p65, MMP-13, Collagen II, SOX9, and IL-1 expression levels. *P<0.05, **P<0.01, ***P<0.001 vs Sham group; ##P<0.01, ###P<0.001 vs Model group.

图5 桃叶珊瑚苷促进软骨细胞修复的细胞实验及软骨细胞模型相关基因表达分析

Fig.5 Alcian blue-Safranin O staining (A, B; Original magnification: ×100) and expressions levels of SOX9, COL2, and TNF-α mRNAs (C, D) in the chondrocyte model. *P<0.05, **P<0.005, ***P<0.001 vs 0 μmol/L.

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桃叶珊瑚苷通过抑制NF-κb信号通路减轻小鼠的膝骨关节炎

Aucubin alleviates knee osteoarthritis in mice by suppressing the NF‑κB signaling pathway

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