山药芪配伍治疗肝细胞癌的机制：网络药理学与实验验证

肝细胞癌（HCC）是全球最常见的恶性肿瘤之一，严重威胁全球公共卫生。免疫细胞功能障碍是HCC免疫抑制微环境形成的关键因素。甘草（Glehnia littoralis (A.Gray) F.Schmidt ex Miq.）与麦冬（Ophiopogon japonicus (Thunb.) Ker Gawl.）是传统中药中滋阴的经典配伍，广泛用于HCC的治疗，具有多重免疫调节功能。然而，山药芪配伍（SS-MD）在HCC管理中的作用及潜在机制尚未阐明。本研究旨在探讨SS-MD配伍治疗HCC的潜在机制。利用UPLC-Q-Orbitrap-MS/MS初步鉴定了SS-MD的已知成分。通过构建药材-成分-靶点网络筛选治疗HCC的活性成分，并通过蛋白质-蛋白质相互作用（PPI）网络探索关键治疗靶点。利用分子对接和分子动力学模拟验证关键靶点与成分的结合亲和力。通过GO生物功能和KEGG通路分析阐明SS-MD配伍治疗HCC的潜在机制，并在肿瘤模型小鼠及细胞共培养实验中验证机制。网络药理学预测显示，SS-MD含有39个活性成分和138个靶点，主要涉及Notch信号通路和细胞凋亡信号通路。靶点富集于淋巴细胞效应功能和淋巴细胞凋亡。体内外实验均证明，SS-MD能改善HCC免疫微环境中CD8+ T细胞比例，调节其亚群分布。SS-MD促进CD8+ T细胞分泌IL-2、TNF-α、IFN-γ、Granzyme B和Perforin，并通过调控Notch信号通路抑制细胞凋亡。该研究识别了SS-MD的关键成分、靶点和信号通路，确认其在HCC治疗中的免疫调节作用，为中医药联合治疗肝癌提供理论依据。

Hepatocellular carcinoma (HCC) is among the most prevalent malignant tumors globally and represents a significant public health issue worldwide. Immune cell dysfunction is the crucial factor for the formation of immunosuppression microenvironment of HCC. Glehnia littoralis (A.Gray) F.Schmidt ex Miq. (Shashen) and Ophiopogon japonicus (Thunb.) Ker Gawl. (Maidong) are classic herb pair in traditional Chinese medicine (TCM) of nourishing Yin, and is widely applied in the treatment of HCC and possesses multiple immunomodulatory functions. However, the role of the Shashen-Maidong herb pair (SS-MD) for the management of HCC and the potential mechanisms has not been explicated.The purpose of the research is to investigate the potential mechanism of the SS-MD herb pair for the management of HCC.The known components of the SS-MD herb pair were preliminarily identified using UPLC-Q-Orbitrap-MS/MS. The active ingredients of SS-MD herb pair in treating HCC were screened by constructing herb-component-target network, and the key therapeutic targets were explored by constructing a protein-protein interaction (PPI) network. The binding affinity of the key targets and components were validated through molecular docking and molecular dynamics simulations. GO biological function and KEGG pathway analyses were operated to elucidate the potential mechanisms of the SS-MD herb pair for the management of HCC. And the mechanism was verified in the tumor bearing mice model and cell co-culture experiments.Network pharmacology prediction revealed 39 active components and 138 targets of the SS-MD herb pair for the treatment of HCC. KEGG analysis mainly focused on Notch signaling pathway and Apoptosis signaling pathway. The targets were enriched in biological functions of lymphocyte effector function and lymphocyte apoptosis. In vivo and in vitro experiments proved that the SS-MD herb pair can improve the proportion of CD8+T cells in the HCC immune microenvironment, regulate its subgroup distribution. SS-MD herb pair promoted CD8+T cells to secrete IL-2, TNF-α, IFN-γ, Granzyme B and Perforin, and inhibited apoptosis by regulating Notch signaling pathway.This study identified the key components, targets, and signaling pathways of the SS-MD herb pair, confirm that SS-MD herb pair play an immunomodulatory role in treating HCC, provides theoretical support for the collaborative treatment of HCC with TCM.