Integration of Single-Nucleus RNA Sequencing and Network Disturbance to Elucidate Crosstalk Between Multicomponent Drugs and Trigeminal Ganglia Cells in Migraine.

OBJECTIVE To clarify the mechanism of Tou Tong Ning capsule（TTNC） and elucidate crosstalk between multi-components drug and trigeminal ganglia cells in migraine. METHODS We integrated single-cell RNA sequencing data and a quantitative evaluation algorithm of the disturbance of multi-target drugs on the disease network to explore the specific pathology of TTNC for migraine. Cerebrovascular smooth muscle spasmolytic activity experiment was carried out to verify the results of bioinformatics analysis. RESULTS TTNC can effectively regulate the functions of synaptic signaling, inflammation and immune response, and cerebrovascular smooth muscle. For different cell subtypes of trigeminal ganglia, TTNC can significantly disturb the robustness on neuronal cell networks and non-neural networks（fibroblast and vascular cells）, indicating TTNC vasodilation activity of brain vessels and neural regulation activities of various neuro types. Contraction of cerebrovascular smooth muscle of mouse ex vivo confirmed the vasodilation activity of TTNC and active compounds（Emodin, Luteolin and Levistilide A）. And literature mining confirmed the vasospasmolytic activity and neuroprotective effect activity of TTNC. CONCLUSIONS Integrating single-cell data and network disturbance tools provides a new strategy for the MoA of multi-components drug through cell subtyping.