The protective effects of Hua-Feng-Dan against chronic cerebral hypoperfusion-induced cerebral ischemia in rats

Objective: This study aimed to examine the effects of different formulations of Hua-Feng-Dan against chronic cerebral ischemia rats and used RNA-Seq to explore the molecular targets. Methods: 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 clinical dose (0.5 g/kg) of Hua-Feng-Dan recipes in diet for 20 days. Finally, behavioral tests were performed, followed by histopathology. Total RNA from hippocampus was isolated and subjected to RNA-Seq, RT-qPCR, and bioinformatics analysis. Results: The CCH rats exhibited cognition deficits evidenced by Morris water maze and Novel object recognition. The neurons in the hippocampus CA1 area were damaged. Hua-Feng-Dan treatments ameliorated behavioral deficits and improved morphological changes. RNA-Seq revealed distinct gene expression pattern between groups. Differentially expressed genes in CCH rats were attenuated or normalized by Hua-Feng-Dan. Ingenuity pathway analysis of canonical pathways and upstream regulators revealed that Ca2+ overload, neuroinflammation, synaptic plasticity and abnormal IGF metabolism were involved in CCH. The Reduced_H, Original and Nonfermented Hua-Feng-Dan at the clinical dose attenuated or reversed these molecular changes, but Reduced_L group was ineffective. Conclusion: Abnormal ion transport, neuroinflammation, synaptic plasticity and metabolism disruption contributed to cognitive deficit in CCH rats, Hua-Feng-Dan recipes (Reduced_H, Original, Nonfermented) at clinical treatment regimen were all effective in ameliorating CCH-induced abnormalities.