In recent years, hypersensitivity reactions to the Shuanghuanglian injection have attracted broad attention. However, the componential chief culprits inducing the reactions and the underlying mechanisms involved have not been completely defined. In this study, we used a combination of approaches based on the mouse model, human umbilical vein endothelial cell monolayer, real-time cellular monitoring, immunoblot analysis, pharmacological inhibition, and molecular docking. We demonstrated that forsythoside A and forsythoside B contributed to Shuanghuanglian injection-induced pseudoallergic reactions through activation of the RhoA/ROCK signaling pathway. Forsythoside A and forsythoside B could trigger dose-dependent vascular leakage in mice. Moreover, forsythoside A and forsythoside B slightly elicited mast cell degranulation. Correspondingly, treatment with forsythoside A and forsythoside B disrupted the endothelial barrier and augmented the expression of GTP-RhoA, p-MYPT1, and p-MLC2 in a concentration-dependent manner. Additionally, the ROCK inhibitor effectively alleviated forsythoside A/forsythoside B-induced hyperpermeability in both the endothelial cells and mice. Similar responses were not observed in the forsythoside E-treated animals and cells. These differences may be related to the potential of the tested compounds to react with RhoA-GTPγS and form stable interactions. This study innovatively revealed that some forsythosides may cause vascular leakage, and therefore, limiting their contents in injections should be considered.

Cheqianzi Decoction (CQD) is a Traditional Chinese Medicine (TCM) formula comprising four herbs and is recorded in the Ancient Materia Medica “Shengji Zonglu”. Individually, these four herbs have been shown to reduce uric acid (UA) levels, to treat hyperuricemia (HUA), and alleviate kidney damage. However, the therapeutic efficacy of the CQD and related mechanism are not yet clear.

Triptonide (TN), a natural bioactive compound derived from Tripterygium wilfordii with multiple antitumor activities, has a poorly defined exact mechanism in acute myeloid leukemia (AML)—a hematologic malignancy with limited treatment options. This study systematically clarifies TN’s mechanisms in AML through an integrative strategy combining network pharmacology, molecular docking, molecular dynamics simulation, and in vitro/in vivo experiment validation. Predicted TN targets using Swiss Target Prediction and PharmMapper, and AML-associated genes via GeneCards, OMIM, and CTD. Verall, O198 overlapping targets were mapped to build a PPI network using STRING and Cytoscape. Identified hub gene (AKT1, EGFR, HSP90AA1, HSP90AB1, and PIK3R1) using CytoNCA, MCODE, and CytoHubba algorithms. GO and KEGG enrichment analyses highlighted marked enrichment in the PI3K/AKT pathway. TN exhibited high affinity binding to AKT1 (−7.28 kcal/mol) and PIK3R1 (−7.33 kcal/mol), with stable interactions confirmed by molecular dynamics simulations. The GSEA of the DEGs from GEPIA 2 revealed prominent activation of the PI3K/AKT signaling pathway, indicating its key role as a regulator of AML pathogenesis. In vitro, TN dose-dependently suppressed proliferation of multiple AML cell lines induced apoptosis, and downregulated the expression of P-PI3K and P-AKT. The AKT activator SC79 reversed TN-induced suppression in AML cells, validating PI3K/AKT pathway dependency. In vivo, TN significantly inhibited tumor growth in xenograft models without causing organ toxicity in female nude mice. These findings reveal the therapeutic potential of TN against AML through inhibiting the PI3K/AKT axis. With no PI3K/AKT inhibitors targeting AML approved as first-line therapies, TN emerges as a promising candidate for AML treatment, offering a safer natural alternative.