Abstract: Objective  To elucidate the mechanism by which Huatan Anshen decoction inhibits neuronal apoptosis through modulation of the NOD-like receptor pyrin domain-containing protein 3 (NLRP3)/interleukin-18 (IL-18) / nuclear factor-activated B cell κ-light chain (NF-κB) signaling pathway. Methods  The HT22 cell line derived from mouse hippocampal neurons were cultured in vitro, and serum containing Huatan Anshen Decoction was prepared from C57 mice after intragastric administration. The optimal concentration of Huatan Anshen Decoction serum for intervention was determined with the CCK-8 method. Following treatment with different concentrations of drug-containing serum, lipopolysaccharide (LPS) was used to induce apoptosis in HT22 cells. Cell damage and apoptosis were assessed with the CCK-8 assay and Annexin V/PI staining. Western blotting and reverse transcription polymerase chain reaction  was employed to measure the expression levels of NF-κB,apoptosis-associated speck-like protein containing (ASC), NLRP3, and Caspase-1, while enzyme-linked immunosorbent assay (ELISA) was used to detect IL-18 expression. The experiments was repeated after overexpression of NRLP3 to conduct single gene function verification. Results  Serum containing Huatan Anshen Decoction significantly improved LPS-induced cellular damage and apoptosis in HT22 cells while reduced cellular L-lactate dehydrogenase, (L-LDH) levels (P<0.05).Transcriptional expression of proteins related to the NLRP3/IL-18/NF-κB signaling pathway showed significant downregulation(P<0.05). Additionally, it markedly decreased intracellular IL-18 levels. Furthermore, overexpression of NF-κB reversed the inhibitory effect of Huatan Anshen decoction-containing serum on HT22 cell apoptosis (P<0.05). Conclusion  Huatan Anshen Decoction can improve neuronal apoptosis via suppressing the NLRP3/IL-18/NF-κB signaling pathway.

Key words: Huatan Anshen Decoction, HT22 cells, apoptosis, NLRP3/IL-18/NF-κB signaling pathway, inflammasome