Puerarin protects against renal ischemia-reperfusion injury by restoring mitochondrial function and modulating the PI3K/AKT/NF-k B pathway and suppressing inflammatory responses

Background: Renal ischemia-reperfusion injury (RIRI) is a major cause of perioperative acute kidney injury and a driver of chronic kidney disease, in which mitochondrial dysfunction and inflammatory activation create a vicious cycle of oxidative stress, inflammation, and tubular cell death. Puerarin, a natural isoflavone with established cardio- and neuroprotective effects, has not been fully evaluated as a modulator of the mitochondrial-immune axis in RIRI.

Methods: Network pharmacology and RNA Sequencing of mouse kidneys with bilateral RIRI were integrated to identify shared puerarin-RIRI targets and enriched pathways. Molecular docking and 100-ns molecular dynamics simulations were performed to assess puerarin-PI3K binding. In vivo, C57BL/6 mice underwent 45-min bilateral renal ischemia and 24-h reperfusion with or without puerarin pretreatment and/or the PI3K Inhibitor LY294002. In vitro, HK-2 cells were subjected to hypoxia/reoxygenation. Renal injury, Apoptosis, oxidative stress, inflammatory cytokines, mitochondrial ultrastructure, membrane potential, and mitochondrial dynamics proteins (Drp1, Opa1, Mfn2) were evaluated.

Results: Network pharmacology and transcriptomics consistently pointed to the PI3K/Akt/NF-κB pathway, which was enriched in oxidative stress, Apoptosis, inflammatory signaling, and mitochondrial processes. In both RIRI mice and HR-treated HK-2 cells, puerarin improved renal histology and function, reduced tubular Apoptosis and IL-6/IL-1β/TNF-α levels, restored SOD and GSH-Px, lowered ROS and MDA, and preserved mitochondrial membrane potential and ultrastructure. These benefits were accompanied by activation of PI3K/Akt signaling and attenuation of NF-κB, together with normalization of mitochondrial fission-fusion (decreased Drp1, increased Opa1/Mfn2), whereas LY294002 markedly blunted puerarin-induced improvements in mitochondrial dynamics, membrane potential, and injury indices.

Conclusion: Puerarin ameliorates RIRI by activating PI3K/Akt signaling, restoring mitochondrial homeostasis, and attenuating NF-κB-linked inflammatory and apoptotic responses. These findings support puerarin as a mitochondria- and immune-targeted candidate for perioperative renal protection and justify further translational evaluation.

Inflammation; Mitochondrial homeostasis; PI3K/AKT/NF-κB pathway; Puerarin; Renal ischemia–reperfusion injury.