Arisaema cum Bile Mitigate Febrile Seizure in Rats via Inhibition of Neuroinflammation, Regulation of FXR and GABA Signaling Pathway

Xu Wu,1,2 Lian Zhong,3 Jing Wang,1,4,5 Qiao Zhang,1,4,5 Jing Sun,1,4,5 Changli Wang,1,4,5 Mengmeng Zhang,1 Chongbo Zhao1,4,5 1College of Pharmacy, Shaanxi University of Chinese Medicine, Xianyang, 712046, People’s Republic of China; 2School of Chinese Materia Medica, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, People’s Republic of China; 3Sichuan Provincial Institute for Drug Control, Chengdu, 611137, People’s Republic of China; 4Shaanxi Provincial Engineering Technology Research Center for Traditional Chinese Medicine Decoction Pieces, Xianyang, 712046, People’s Republic of China; 5Traditional Chinese Medicine Processing Techniques Heritage Base (Shaanxi), National Administration of Traditional Chinese Medicine, Xianyang, 712046, People’s Republic of ChinaCorrespondence: Mengmeng Zhang, College of Pharmacy, Shaanxi University of Chinese Medicine, Qindu District, Xianyang, 712046, People’s Republic of China, Email [email protected] Chongbo Zhao, College of Pharmacy, Shaanxi University of Chinese Medicine, Qindu District, Xianyang, 712046, People’s Republic of China, Email [email protected]: Arisaema cum bile (known as DanNan Xing in Chinese, DNX) is a traditional herbal medicine commonly used to treat febrile seizure (FS), but the underlying mechanism remains unclear.Objective: To evaluate the therapeutic effect of DNX on hot water bath-induced FS rat model and further explore the potential mechanism.Methods: The chemical constituents of DNX were determined via liquid chromatography-mass spectrometry (LC-MS). FS rat model was established using a hot water bath (45 ± 2 °C), and DNX (2.8 and 0.7 g/kg, i.g) were administered for two weeks. Based on behavior test (duration and latency), pathological changes in the hippocampal tissue, and the levels of inflammatory cytokines the therapeutic effect of DNX for FS was evaluated. Subsequently, the network pharmacology, 16S rRNA and non-targeted metabolomics analysis were combined analysis to explore the potential signaling pathway. Furthermore, the signaling pathway was verified using the RT-qPCR and immunohistochemistry assay.Results: The DNX treatment showed effective therapy on hot water bath induced FS, as indicated by a shortened seizure duration time, prolonged seizure latency, reduced hippocampal neuron damage and neuroinflammatory factor levels (TNF-α, IL-1β, IL-6, and HMGB1). Neurotransmitters (GABA, Glu) are also significantly regulated. Moreover, the relative abundance of Lactobacillus and Lachnospiraceae was notably increased (p < 0.01), while that of Tenericutes decreased, compared to gut microbiota of FS rat. A total of 20 fecal differential metabolites were regarded as the potential biomarkers including GABA, CDCA, and UDCA for anti-FS of DNX, and combined network pharmacy the metabolic pathways of primary bile acids (BAs) biosynthesis and alanine, aspartate and glutamate metabolism were involved.Conclusion: DNX possesses a therapeutic effect on FS through inhibiting neuroinflammation and regulation of FXR and GABA signaling pathway.Keywords: Arisaema cum bile, febrile seizures, neuroinflammation, gut microbiota, fecal metabolites, FXR and GABA signaling