The protection of chronic cerebral ischemia by Hua-Feng-Dan involves the modulation of hippocampal transcriptome and gut dysbiosis

The protective effects of Hua-Feng-Dan recipes against chronic cerebral hypoperfusion-induced ischemia were examined; RNA-seq and 16 S-rRNA-seq were used to explore the molecular mechanisms. Male Sprague-Dawley rats underwent unilateral common carotid artery ligation surgery to establish chronic cerebral hypoperfusion (CCH) models. Then, the CCH rats were treated with various Hua-Feng-Dan recipes in diet for 20 days. Finally, behavioral tests were performed, followed by histopathology. Hippocampal RNA was isolated for RNA-seq and RT-qPCR analyses; Colon bacterial DNA was isolated for 16 S rRNA-seq and bioinformatics. The CCH rats exhibited cognition deficits evidenced by Behavioral tests. The neurons in the hippocampus CA1 area were damaged. Hua-Feng-Dan recipes as Reduced_H (0.5 g/kg), Original, and Nonferment improved behavioral deficits and morphological changes, while Reduced_L (0.25 g/kg) was ineffective. RNA-seq revealed distinct gene expression pattern among groups. Ingenuity pathway analysis of differentially expressed genes revealed Hua-Feng-Dan recipes ameliorated Canonical pathways involved in cytosolic Ca 2+ overload, disruption of IGF homeostasis and Neutrophil degranulation; attenuated CCH-activated Upstream regulators such as IL1B, IL6, TNF, IGF1, and glucose metabolism. CCH-decreased Firmicutes/Bacteroidetes ratio were recovered by Hua-Feng-Dan recipes. CCH-increased Prevotellaceae and decreased Lachnospiraceae were attenuated; CCH-increased ASV1 and decreased ASV39 were brought to normal after Hua-Feng-Dan treatments. Similar to the efficacy against CCH, Reduced_L was ineffective in transcriptome and gut microbiota analysis. Hua-Feng-Dan recipes (Reduced_H, Original, and Nonferment) are effective against CCH-induced chronic cerebral ischemia, while Reduced_L was ineffective. The mechanisms of the protection are related to transcriptome regulation in the hippocampus, and modulation of CCH-disrupted gut dysbiosis.