Network Pharmacology, Experimental Validation Explored Kakkalide to Ameliorate Endothelial Cell Dysfunction and Inflammatory Response.

BACKGROUND: Sepsis continues to represent a significant challenge due to its detrimental effects and high mortality rate. The protection of endothelial function and the attenuation of the excessive inflammatory response are pivotal in the reduction of mortality risk. Flos Puerariae is the flower of Pueraria montana var. lobata (Willd). Flos Puerariae was recorded as ameliorating loathing of cold with high fever and clearing heat and purging the lung from "Minnan Materia Medica". Recent findings showed its potential efficacy of inflammatory diseases. However, the precise underlying mechanisms are produced remain to be elucidated. PURPOSE: This study aimed to identify the key active compound in Flos Puerariae for sepsis treatment and its pharmacological targets. METHODS: Network pharmacology analysis identified potential targets of Flos Puerariae against sepsis. Cecal ligation and puncture (CLP) mouse models assessed survival, inflammation, and tissue damage via ELISA and H&E staining. Transwell assays, immunofluorescence (IF), and flow cytometry evaluated human umbilical vein endothelial cell (HUVEC) protection. Molecular docking and Western blot (WB) confirmed key signaling pathway targets. RESULTS: At a dose of 20 mg/kg, the Flos Puerariae-derived compound Kakkalide notably reduced mortality and serum inflammatory factor levels while protecting lung and renal tissues. In vitro experiments further showed that treatment with 5 μM Kakkalide in HUVEC decreased inflammatory factors, reactive oxygen species, and apoptosis, while promoting cell migration and proliferation. The aforementioned effects were associated with alterations in the phosphorylation levels of Tumor Necrosis Factor (TNF), extracellular regulated protein kinases (ERK), and protein kinase B (AKT). CONCLUSION: Our study demonstrated that Kakkalide as the primary ingredient of Flos Puerariae enhanced the survival of CLP mice and protected lipopolysaccharides (LPS)-stimulated HUVEC function. This study offers a novel perspective on the effects and molecular mechanisms of Kakkalide in sepsis.