PH/ROS Dual-Responsive Mitochondria-Targeting Self-Assembled Nanoparticles Loaded with Phillyrin Attenuate Sepsis-Associated Acute Kidney Injury

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PH/ROS Dual-Responsive Mitochondria-Targeting Self-Assembled Nanoparticles Loaded with Phillyrin Attenuate Sepsis-Associated Acute Kidney Injury

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Co-author 1

Jia Chen 664816914@qq.com Army Medical University Department of Nephrology, Daping Hospital Chongqing China *

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Kehong Chen chenkehong@foxmail.com Army Medical University Department of Nephrology, Daping Hospital Chongqing China -

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Introduction

Sepsis-associated acute kidney injury (SA-AKI) is a complex and life-threatening clinical syndrome, but effective treatment remains challenging.

Methods

Proteomic analysis and validation experiments to study the mechanism of SIRT3 in SA-AKI. Phillyrin (PHL), a bioactive glycoside compound found in Forsythia suspensa fruit, was proven to be a natural agonist of SIRT3. To enhance renal mitochondrial targeting, we developed novel PHL-loaded nanoparticles (NPs) for SA-AKI therapy. The amino group (-NH2) of the tert butoxycarbonyl ketothiol amino group (NH2-TK-BoC, ROS-responsive) reacted with the aldehyde group (-CHO) of the triphenyl phosphate polyethylene glycol aldehyde group (TPP-PEG-CHO, mitochondrial targeting) to form a Schiff bond (TPP-PEG-C=N-TK-BoC, pH-responsive) through a Schiff base reaction. This compound was further conjugated with a renal tubular epithelial cell-specific peptide (KKEEE), yielding TPP-PEG-C=N-TK-BoC-KKEEE (TCTK) NPs through self-assembly. These NPs were then loaded with PHL to form PTCTK NPs. Characterization tests confirmed the successful construction of PTCTK NPs. The effects of PTCTK NPs loaded with phillyrin were demonstrated in both SA-AKI mouse models and LPS-stimulated HK-2 cells.

Results

Proteomic analysis and validation studies revealed decreased expression of SIRT3 and NRF3. Mechanistic research revealed that reduced SIRT3 binds to NRF3 and promotes its acetylation, which results in inflammation and mitochondrial damage.Characterization tests confirmed the successful construction of PTCTK NPs. In both SA-AKI mouse models and LPS-stimulated HK-2 cells, PTCTK NPs effectively targeted renal mitochondria and resulted in dual ROS/pH responsiveness. PTCTK NPs reduced inflammation and oxidative stress by modulating SIRT3/NRF3 signaling, thereby alleviating SA-AKI.

Conclusion

These findings suggest that the novel PTCTK NPs loaded with phillyrin could effectively improve SA-AKI and may serve as a promising nanotherapeutic strategy for SA-AKI.

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