Andrographolide prevents necroptosis by suppressing the generation of reactive oxygen species

• Andrographolide (Andro) effectively suppresses necroptosis in vitro by inhibiting the RIPK1/RIPK3/MLKL signaling pathway. • Andro reduces intracellular ROS and mitochondrial superoxide, preserves mitochondrial membrane potential, and activates Nrf2, a key antioxidant transcription factor. • Andro prevents the oligomerization and co-localization of mitochondrial proteins (Bcl-2, Bak) with necrosome components, thereby blocking necrosome formation. • Structural derivatives of Andro lack anti-necroptotic activity and fail to upregulate Nrf2, indicating specificity of Andro's mechanism.