Constructing a Preeclampsia Organoid Model to Elucidate the Mechanism of Aspirin

BACKGROUND: Preeclampsia is a life-threatening pregnancy disorder characterized by hypertension and multiorgan dysfunction, posing significant risks to both maternal and fetal health. Although low-dose aspirin is widely recommended for preventing preeclampsia, the underlying mechanisms of action are still poorly understood, which hinders the optimization of therapeutic strategies. METHODS: We developed an in vitro hypoxia-induced preeclampsia model using human trophoblast organoids to replicate key pathological features. RNA sequencing identified dysregulated pathways and molecular targets. Functional assays assessed the effects of aspirin on trophoblast proliferation, mitochondrial activity, and hormonal regulation, focusing on the PI3K (phosphatidylinositol 3-kinase)-AKT (protein kinase B) pathway and CYP (cytochrome P450) enzymes. We also analyzed the effects of aspirin in the N'-nitro-L-arginine-methyl ester hydrochloride rat model. RESULTS: The hypoxia-induced preeclampsia model successfully mimicked clinical hallmarks, including elevated sFLT-1 (soluble fms-like tyrosine kinase 1)/PlGF (placental growth factor) ratios and oxidative damage. RNA sequencing revealed significant suppression of the PI3K-AKT-mTOR (mammalian target of rapamycin) pathway and dysregulation of CYP enzymes. Aspirin treatment restored the sFLT-1/PlGF balance, reactivated the PI3K-AKT-mTOR pathway, and improved mitochondrial function, enhancing trophoblast proliferation. Furthermore, aspirin regulated CYP by increasing CYP19A1 (cytochrome P450 1A1) and inhibiting CYP1A1 (cytochrome P450 1A1), thereby improving placental hormonal homeostasis. CONCLUSIONS: This study clarifies aspirin's multitarget mechanisms in alleviating preeclampsia, which include restoring the sFLT-1/PlGF balance, activating the PI3K-AKT-mTOR signaling pathway, optimizing mitochondrial function, and regulating CYP-mediated hormonal metabolism. These findings provide a mechanistic basis for aspirin's clinical effectiveness in preventing preeclampsia.