Single Cell Transcriptomics of Traditional Chinese Medicine Bioactives: Mapping Immune, Neural and Hepatic Responses.

Abstract

Traditional Chinese medicine (TCM) bioactives display wide pharmacological effects, yet bulk transcriptomic studies mask their cell-specific actions. This study applied single-cell RNA sequencing to delineate immune, neural and hepatic responses to chemically characterized TCM constituents. Murine splenic, hippocampal and hepatic tissues were dissociated into single-cell suspensions, processed on a 10× Genomics platform, sequenced at high depth and analyzed with Seurat for clustering, marker annotation, differential expression and pathway enrichment; ligand-receptor interactions were inferred by CellChat, with RT-qPCR and immunofluorescence validation. From over 42,000 high-quality cells, we resolved 15 immune, 12 neural and 10 hepatic subtypes. TCM exposure induced NF-κB modulators in macrophages, neurotrophic gene expression in astrocytes and xenobiotic-metabolism programs in hepatocytes, while pathway analysis highlighted coordinated immune-hepatic detoxification and neuroprotective signaling. Ligand-receptor mapping revealed strengthened IL-10-STAT3 and hepatocyte-Kupffer cross-talk. These findings demonstrate that single-cell transcriptomics exposes previously hidden, cell-type-specific pharmacodynamics of TCM bioactives, laying a mechanistic foundation for precision-oriented herbal pharmacology and rational drug development.