School of Chinese Pharmacy, Beijing University of Chinese Medicine, 102488, Beijing, China.
College of Traditional Chinese Medicine, Xinjiang Medical University, 830011, Urumqi, China.
Cell Biochem Biophys. 2023 Sep;81(3):515-532. doi: 10.1007/s12013-023-01151-2. Epub 2023 Jul 31.
We investigated the mechanisms underlying the effects of Ziziphora clinopodioides Lam. (ZCL) on atherosclerosis (AS) using network pharmacology and in vitro validation.We collected the active components of ZCL and predicted their targets in AS. We constructed the protein-protein interaction, compound-target, and target-compound-pathway networks, and performed GO and KEGG analyses. Molecular docking of the active components and key targets was constructed with Autodock and Pymol software. Validation was performed with qRT-PCR, ELISA, and Western blot.We obtained 80 components of ZCL. The network analysis identified that 14 components and 37 genes were involved in AS. Then, 10 key nodes in the PPI network were identified as the key targets of ZCL because of their importance in network topology. The binding energy of 8 components (Cynaroside, α-Spinasterol, Linarin, Kaempferide, Acacetin, Genkwanin, Chrysin, and Apiin) to 4 targets (MMP9, TP53, AKT1, SRC) was strong and <-1 kJ/mol. In addition, 13 of the 14 components were flavonoids and thus total flavonoids of Ziziphora clinopodioides Lam. (ZCF) were used for in vitro validation. We found that ZCF reduced eNOS, P22, gp91, and PCSK9 at mRNA and protein levels, as well as the levels of IL-1β, TNF-α, and IL-6 proteins in vitro (P < 0.05).We successfully predicted the active components, targets, and mechanisms of ZCL in treating AS using network pharmacology. We confirmed that ZCF may play a role in AS by modulating oxidative stress, lipid metabolism, and inflammatory response via Cynaroside, Linarin, Kaempferide, Acacetin, Genkwanin, Chrysin, and Apiin.
我们使用网络药理学和体外验证研究了香青兰（ZCL）治疗动脉粥样硬化（AS）的作用机制。我们收集了 ZCL 的活性成分，并预测了其在 AS 中的靶点。我们构建了蛋白质-蛋白质相互作用、化合物-靶点和靶点-化合物-通路网络，并进行了 GO 和 KEGG 分析。使用 Autodock 和 Pymol 软件构建了活性成分和关键靶点的分子对接。采用 qRT-PCR、ELISA 和 Western blot 进行了验证。我们获得了 ZCL 的 80 种成分。网络分析确定了 14 种成分和 37 个基因参与了 AS。然后，由于在网络拓扑中的重要性，PPI 网络中的 10 个关键节点被确定为 ZCL 的关键靶点。8 种成分（梓醇苷、α-菠菜甾醇、木犀草素、山柰酚、芹菜素、根皮苷、白杨素、和 Apiin）与 4 个靶点（MMP9、TP53、AKT1、SRC）的结合能较强，均<-1 kJ/mol。此外，14 种成分中的 13 种为类黄酮，因此我们使用香青兰总黄酮（ZCF）进行了体外验证。我们发现 ZCF 降低了 eNOS、P22、gp91 和 PCSK9 的 mRNA 和蛋白水平，以及体外 IL-1β、TNF-α 和 IL-6 蛋白水平（P < 0.05）。我们成功地使用网络药理学预测了 ZCL 治疗 AS 的活性成分、靶点和机制。我们通过 Cynaroside、木犀草素、山柰酚、芹菜素、根皮苷、白杨素和 Apiin 证实了 ZCF 可能通过调节氧化应激、脂质代谢和炎症反应在 AS 中发挥作用。
