The acute oral toxicity in rats fed technical grade azadirachtin ranged from greater than 3,540 mg/kg to greater than 5,000 mg/kg, the highest dose tested when administered undiluted to albino rats. The single-dose oral toxicity LD50 of the formulated product Azatin-EC fed to rats was 4,241 mg/kg; considered practically nontoxic

Azadirachtin (AZA) primarily induced autophagy in Spodoptera litura cultured cell line (SL-1 cell) by dysregulating InR- and PI3K/AKT/TOR pathways, then stimulated apoptosis by activating tAtg5. The PI3K/AKT/TOR signaling pathway was investigated in SL-1 cells to explore the molecular mechanism underlying the autophagy induced by AZA. The PI3K signaling pathway negatively controls autophagy. The results demonstrated decreased PI3K (85KD) in SL-1 cells following treatment with 2.5 μg/mL AZA for 24 h. It was also evaluated the AZA cytotoxicity using the WST-8 test to verify whether AZA was capable of inducing autophagy in SL-1 cells. The results showed that SL-1 cell proliferation was inhibited in a dose-dependent manner after 24 h exposure. The inhibition activity can be significantly observed at a low concentration of 0.625 μg/mL. The cell viability rates were 80.19%, 74.72%, 69.41%, 67.43%, 66.22%, 63.83% and 49.84% at the concentrations of 1.25, 2.5, 5, 10, 20, 40 and 80 μg/mL, respectively.