Potential targets of baicalein in macrophages revealed by bulk and single cell RNA sequencing analysis

Excessive inflammation drives organ dysfunction and high mortality in life-threatening conditions such as sepsis. Baicalein, a bioactive flavonoid, possesses well-recognized anti-inflammatory properties, yet its molecular targets in macrophages and potential systemic immunomodulatory effects on peripheral blood immune cells under hyperinflammatory conditions remain poorly characterized. Our previous studies demonstrated that baicalein alleviates hepatic inflammation in mice with non-alcoholic fatty liver disease (NAFLD) and inhibits NF-κB nuclear translocation in RAW264.7 macrophages. Here, by integrating network pharmacology, molecular docking, bulk RNA sequencing of macrophages, and single-cell RNA sequencing of peripheral blood from sepsis patients, we identified JAK2, SRC, TP53, MAPK3, AKT1, HSP90AA1, and ESR1 as potential core targets of baicalein in macrophages, and validated that the JAK2-STAT3 and NF-κB pathways might be the key downstream regulatory axes of its anti-inflammatory effects. Furthermore, we revealed that baicalein may modulate, based on single-cell expression signatures, the inflammatory phenotype of multiple peripheral blood immune cell populations, including monocytes, T cells, B cells, and granulocyte-monocyte progenitors, suggesting a potential systemic anti-inflammatory effect that requires experimental validation in human cells. Collectively, our findings elucidate the potential molecular targets of baicalein in macrophages and its multi-cellular immunoregulatory mechanisms under hyperinflammation, providing novel mechanistic insights for the clinical application of baicalein in inflammatory diseases.