Antiepileptic Effects of Hua-Feng-Dan Against Pentylenetetrazol-Induced Seizures in Mice.

Abstract

Hua-Feng-Dan is a Chinese medicine in the treatment of epilepsy and neurodegeneration. The prior studies demonstrated its protection against neuroinflammation, dopaminergic neuron loss, and in the treatment of ischemic stroke, but its anticonvulsive efficacy against epilepsy remains elusive. The aim of this study was to examine antiepileptic effects of Hua-Feng-Dan recipes and the mechanism of protection. Pentylenetetrazol (PTZ)-induced kindling mouse model (35 mg/kg, ip on alternate days × 10) was used to evaluate the antiepileptic effects of Hua-Feng-Dan recipes. Mice were given original, reduced, and nonfermented Hua-Feng-Dan via feed at clinical dose (0.5 g/kg) and diazepam was used as a positive control. The seizure scores were recorded after each PTZ injection. At the end of the experiments, the brain and colon content were collected for RNA-Seq and 16S rRNA-Seq, respectively, followed by bioinformatics and qPCR verification. Hua-Feng-Dan recipes were effective against PTZ-induced seizures, with original the best and half of reduced (0.25 g/kg) ineffective. PTZ-induced aberrant gene expressions in the brain were ameliorated by Hua-Feng-Dan recipes to various extents. Ingenuity Pathway Analysis of differentially expressed genes revealed that PTZ-altered canonical pathways and upstream regulators were attenuated by Hua-Feng-Dan treatments. qPCR verified the expression of early immediate genes, proinflammatory mediators, transporters and apoptosis genes. PTZ-disrupted gut microbiome was also ameliorated by Hua-Feng-Dan recipes, and the modulation of Lachnospiraceae and ASV230 was consistent with the literature. Hua-Feng-Dan recipes were effective against PTZ-induced kindling mice. The mechanisms appeared to be related to the modulation of brain gene expression and gut microbiota. • Hua-Feng-Dan (HFD) recipes were effective against PTZ-induced seizures in mice. • HFD ameliorated abnormal brain gene expressions in PTZ kindling mice via RNA sequencing (RNA-seq). • IPA of DEGs revealed improved canonical pathways and upstream regulators. • 16S rRNA-seq showed microbiome modulation by HFD in PTZ kindling mice. • qPCR verified selected brain genes and bacterial ASVs.