Neem leaf extract alleviates LPS/D-GalN induced acute hepatitis in mice through its anti-inflammatory effects and activation of autophagy

Acute hepatitis, characterized by rapid onset and high mortality, can result from infections, toxins, and other factors. However, current treatment options have significant side effects, necessitating further research into alternative therapies. This study investigated the extraction method of neem extract and found that its ethanolic extract effectively reduced mortality and decreased ALT and AST levels in mice serum, improving liver pathology. HPLC analysis identified azadirachtin and nimbolide in the extract. It also downregulated NF-κB, NLRP3, and p62 levels, while upregulating Lc3B and Atg5 levels. Experiments in Atg5 knockout mice showed that the absence of Atg5 weakened the extract's efficacy in reducing liver damage and inflammation and affected the extent of NLRP3 protein downregulation. However, it did not affect the extract's ability to reduce NF-κB. Overall, the ethanolic extract of neem leaves primarily modulates the inflammatory response through the NF-κB signaling pathway. The extract's efficacy in reducing NLRP3 is associated with autophagy. These discoveries offer a new theoretical basis for the role of neem in treating acute hepatitis.