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| 补肾壮膝方治疗膝骨关节炎的作用机制研究 |
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金振辉1, 皮林君2, 谢智玖2, 董俐珑1, 林颖焘1, 邵丹玥1, 王玉英1, 王景霞1
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1.北京中医药大学中医学院, 北京 100029;2.新疆医科大学中医学院, 乌鲁木齐 830017
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| 摘要: |
| [目的] 基于超高效液相色谱-四极杆-飞行时间质谱（UPLC-Q-TOF-MS）技术、网络药理学和动物实验，探讨补肾壮膝方治疗肾阳虚型膝骨关节炎（KOA）的潜在分子机制。[方法] 采用 UPLC-Q-TOF-MS 对补肾壮膝方的化学成分进行定性分析。通过 Swiss Target Prediction 等数据库获取成分靶点，与 KOA 疾病靶点取交集，构建蛋白-蛋白相互作用（PPI）网络，进行 GO 和 KEGG 富集分析。采用卵巢切除联合关节腔注射建立肾阳虚 KOA 大鼠模型，随机分为假手术组、模型组和补肾壮膝方低、中、高剂量组。给药 28 d 后，观察大鼠肾阳虚证候、痛阈和膝关节肿胀度；ELISA 检测血清和关节液中相关指标；qRT-PCR 检测关节软骨组织中磷脂酰肌醇 3-激酶（PI3K）-蛋白激酶 B（AKT）与核因子-κB（NF-κB）通路关键基因的表达。[结果] 从补肾壮膝方中鉴定出 74 个化合物。网络药理学筛选出 122 个药物-疾病共有靶点，PPI 网络显示肿瘤坏死因子（TNF）、白细胞介素 6（IL6）、蛋白激酶 B1（AKT1）等是关键靶点，KEGG 富集分析提示 PI3K-AKT 和 NF-κB 信号通路是核心通路。动物实验结果表明，补肾壮膝方能显著改善肾阳虚证候，提高痛阈，减轻软骨损伤。qRT-PCR 结果显示，与空白组相比，模型组大鼠软骨组织中 PI3K、AKT、NF-κB p65、IκBα 的 mRNA 表达均显著升高（P<0.05 或 P<0.001）；与模型组相比，补肾壮膝方高剂量组可显著下调上述所有基因的 mRNA 表达（P<0.05 或 P<0.001）。[结论] 补肾壮膝方能够改善肾阳虚型 KOA 大鼠的症状，延缓软骨退变。其机制可能与调控 PI3K-AKT 与 NF-κB 通路的串扰有关，体现了中药多成分、多靶点、多通路的作用特点。 |
| 关键词: 补肾壮膝方 膝骨关节炎 UPLC-Q-TOF-MS 网络药理学 PI3K-AKT通路 NF-κB通路 |
| DOI：10.11656/j.issn.1672-1519.2026.10.13 |
| 分类号:R285.5 |
| 基金项目:北京中医药薪火传承“新3+3”工程王玉英三名传承工作室（2023-SZ-F-22）。 |
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| Study on the mechanism of Bushen Zhuangxi Formula in the treatment of knee osteoarthritis |
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JIN Zhenhui1, PI Linjun2, XIE Zhijiu2, DONG Lilong1, LIN Yingtao1, SHAO Danyue1, WANG Yuying1, WANG Jingxia1
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1.School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China;2.School of Traditional Chinese Medicine, Xinjiang Medical University, Urumqi 830017, China
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| Abstract: |
| [Objective] To explore the potential molecular mechanism of Bushen Zhuangxi Formula（BSZXF）in the treatment of knee osteoarthritis（KOA）with kidney-yang deficiency based on ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS）technology，network pharmacology and animal experiments. [Methods] UPLC-Q-TOF-MS was used to qualitatively analyze the chemical composition of BSZXF. Swiss Target Prediction and other databases were used to obtain the component targets，and the intersections with KOA disease targets were taken to construct the protein-protein interaction（PPI） network，and GO and KEGG enrichment analysis were performed. The KOA rat model of kidney-yang deficiency was established by ovariectomy combined with intra-articular injection. The rats were randomly divided into sham operation group，model group，and BSZXF low，medium，and high-dose groups. After 28 days of administration，the syndrome of kidney-yang deficiency，pain threshold and knee joint swelling degree were observed. ELISA was used to detect related indicators in serum and synovial fluid. The qRT-PCR was used to detect the expression of key genes in PI3K-AKT and NF-κB pathways in articular cartilage tissue. [Results] Seventy-four compounds were identified from BSZXF. PPI network showed that TNF，IL6 and AKT1 were the key targets. KEGG enrichment analysis showed that PI3K-AKT and NF-κB signaling pathways were the core pathways. Animal experiments have shown that BSZXF can significantly improve the syndrome of kidney-yang deficiency，increase the pain threshold，and reduce cartilage damage. qRT-PCR results showed that compared with the blank group，the mRNA expressions of PI3K，AKT，NF-κB p65，and IκBα in cartilage tissue of the model group were significantly increased（P<0.05 or P<0.001）. Compared with the model group，the high-dose BSZXF group significantly down-regulated the mRNA expression of all the above genes（P<0.05 or P<0.001）. [Conclusion] BSZXF can improve the symptoms of KOA rats with kidney-yang deficiency and delay cartilage degeneration. The mechanism may be related to the crosstalk between PI3K-AKT and NF-κB pathways，which reflects the characteristics of multi-components，multi-targets and multi-pathways of traditional Chinese medicine. |
| Key words: Bushen Zhuangxi Formula knee osteoarthritis UPLC-Q-TOF-MS network pharmacology PI3K-AKT pathway NF-κB pathway |
