| 摘要: |
| [目的] 基于网络药理学及实验验证探讨芪麝丸及其有效成分延缓椎间盘退变及髓核细胞衰老的机制。[方法] 基于网络药理学筛选芪麝丸活性成分及其作用靶点，构建“成分-靶点-通路”网络；通过分子对接验证芪麝丸有效成分与相关信号通路的关键靶点结合能力；结合体外实验，构建髓核细胞衰老模型，通过蛋白质免疫印迹及免疫荧光等实验确定有效单体并探究其作用通路的上下游关系。[结果] 网络药理学交集靶点明显富集于磷酸化蛋白53（p53）衰老信号通路，且与多条相关通路联系密切；分子对接及实验验证筛选出汉防己甲素为芪麝丸延缓椎间盘退变的有效成分；体外实验证实汉防己甲素可通过上游磷酸肌醇3-激酶（PI3K）通路抑制p53通路活性，减少髓核细胞凋亡并促进胶原蛋白Ⅱ型（COL2A1）合成。[结论] 髓核细胞衰老是椎间盘退变的关键病理因素，研究揭示了芪麝丸延缓椎间盘退变的机制，并筛选出其有效成分汉防己甲素，证明其通过调控PI3K通路延缓椎间盘退变和髓核细胞衰老的作用。 |
| 关键词: 椎间盘退变 芪麝丸 网络药理学 p53信号通路 体外实验 |
| DOI：10.11656/j.issn.1673-9043.2026.06.10 |
| 分类号:R681.5 |
| 基金项目:国家自然科学基金面上项目（82474536）；上海市卫生健康委员会临床研究专项面上项目（202440110）；2024年度江苏省中医药科技发展计划青年人才项目（QN202419）；2024年度苏州市应用基础研究科技创新项目（SYWD2024154）；上海市第六人民医院院级科研基金项目（ynhg202420）。 |
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| Exploring the mechanism of Qishe Pills and its active component in delaying intervertebral disc degeneration and nucleus pulposus cell senescence：A study based on network pharmacology and experimental validation |
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ZHANG Cheng1, WANG Bingquan2, DAI Yuxiang3, WANG Yongjun1, SUN Yueli1
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1.Spine Disease Institute, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China;2.Department of Acupuncture and Moxibustion Tuina Traumatolog, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200032, China;3.Department of Orthopedics, Suzhou TCM Hospital affiliated to Nanjing University of Traditional Chinese Medicine, Suzhou 215009, China
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| Abstract: |
| [Objective] To investigate the mechanism by which Qishe Pills（QSP） and its effective components delay intervertebral disc degeneration（IVDD） and nucleus pulposus（NP） cell senescence，utilizing network pharmacology and experimental validation. [Methods] Active components of QSP and their potential targets were screened based on network pharmacology，and a “Component-Target-Pathway” network was constructed. The binding affinity between active components of QSP and key targets of relevant signaling pathways was validated by molecular docking. Combined with in vitro experiments，an NP cell senescence model was established. Effective monomers were identified，and the upstream-downstream relationships of their action pathways were explored using Western blot and immunofluorescence assays. [Results] Network pharmacology revealed that the intersection targets were significantly enriched in the p53 signaling pathway and were associated with the PI3K/AKT，AMPK/SIRT1，ERK，and TGF-β/Smad pathways. Molecular docking and experimental validation identified tetrandrine as the effective component of QSP regulating IVDD. In vitro experiments confirmed that tetrandrine inhibits p53 pathway activity via the upstream PI3K/AKT/MDM2 pathway，thereby reducing NP cell apoptosis and promoting the synthesis of type II collagen（COL2A1）. [Conclusion] NP cell senescence is a key pathological factor in IVDD. This study elucidates the mechanism by which QSP delay IVDD，identifies tetrandrine as its effective component，and demonstrates that tetrandrine mitigates IVDD and NP cell senescence by regulating the PI3K/AKT signaling pathway. |
| Key words: intervertebral disc degeneration Qishe Pills network pharmacology p53 signaling pathway in vitro experiments |
