-Mediated Insulin Signaling Pathway Was Involved in O-Induced Multigenerational Effects of Shortened Lifespan in .

As a long-standing atmospheric pollutant, ozone (O) exerts enduring effects on biological health. However, experimental research on its impact on organism lifespan and generational effects is limited. This study exposed three generations of fruit flies () to O, revealing a shortened lifespan across generations. Specifically, after O exposure, the lifespan of the F2 generation was significantly reduced compared with F0 and F1 generations, indicating a cumulative multigenerational effect. Transcriptome analysis unveiled significant disruptions in metabolic pathways, notably the insulin signaling pathway. Subsequent qRT-PCR analysis showed elevated mRNA levels of insulin pathway-related genes (, , , , and ), alongside decreased expression levels of , , and in flies exposed to O. Notably, knocking down , rather than , and , rescued the O-induced lifespan shortening. Overall, O exposure triggered activation of the dilp2-mediated InR-FOXO/TOR-4E-BP-Atg5 signaling pathway, potentially contributing to shortened lifespan with cumulative effects. This study highlights the viability of employing fruit flies as a model to evaluate the multigenerational toxicity of environmental pollutants, particularly atmospheric pollutants.