Enhanced protection against hypoxia/reoxygenation-induced apoptosis in H9c2 cells by puerarin-loaded liposomes modified with matrix metalloproteinases-targeting peptide and triphenylphosphonium

Abstract

Based on the inhibition of mitochondrial permeability transition pore (mPTP) opening, puerarin (PUE) has a good potential to reduce myocardial ischemia/reperfusion injury (MI/RI). However, the lack of targeting of free PUE makes it difficult to reach the mitochondria. In this paper, we constructed matrix metalloproteinase-targeting peptide (MMP-TP) and triphenylphosphonium (TPP) cation co-modified liposomes loaded with PUE (PUE@T/M-L) for mitochondria-targeted drug delivery. PUE@T/M-L had a favorable particle size of 144.9 ± 0.8 nm, an encapsulation efficiency of 78.9 ± 0.6%, and a sustained-release behavior. The results of cytofluorimetric experiments showed that MMP-TP and TPP double-modified liposomes (T/M-L) enhanced intracellular uptake, escaped lysosomal capture, and promoted drug targeting into mitochondria. In addition, PUE@T/M-L enhanced the viability of hypoxia-reoxygenation (H/R) injured H9c2 cells by inhibiting mPTP opening and reactive oxygen species (ROS) production, reducing Bax expression and increasing Bcl-2 expression. It was inferred that PUE@T/M-L delivered PUE into the mitochondria of H/R injured H9c2 cells, resulting in a significant increase in cellular potency. Based on the ability of MMP-TP to bind the elevated expression of matrix metalloproteinases (MMPs), T/M-L had excellent tropism for Lipopolysaccharide (LPS) -stimulated macrophages and can significantly reduce TNF-α and ROS levels, thus allowing both drug accumulation in ischemic cardiomyocytes and reducing inflammatory stimulation during MI/RI. Fluorescence imaging results of the targeting effect using a DiR probe also indicated that DiR@T/M-L could accumulate and retain in the ischemic myocardium. Taken together, these results demonstrated the promising application of PUE@T/M-L for mitochondria-targeted drug delivery to achieve maximum therapeutic efficacy of PUE.

Keywords:

• Mitochondrial targeting
• puerarin
• hypoxia/reoxygenation
• matrix metalloproteinase-targeting peptide
• RAW 264.7 cells

Acknowledgements

We are grateful to Professor Daxiong Xiang for the characterization of materials. We are grateful to all members of the research group for providing language and writing assistance and a conducive work environment.

Author contributions

L-FM, W-Y, L-SN, and L-XY: performed the literature search and design, conducted experiments, analyzed data, and wrote the manuscript. L-WQ, W-JY, H-XB, and T-TT: performed the literature search and conducted experiments. L-XY: conceived and designed the experiments and critically revised the manuscript. All authors discussed the results and contributed to the manuscript writing.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

The datasets generated for this study are available on request to the corresponding author.

Additional information

Funding

Financial support for this work is provided by the National Natural Science Foundation of China (81673614), Hunan Provincial Natural Scientific Foundation (No.2020JJ4128, 2020JJ5808), Scientific Research Project of Hunan Provincial Health and Family Planning Commission (No. 202113020290), Hunan provincial science and technology plan (2016TP2002), Graduate Independent Exploration and Innovation Project of Central South University (2022ZZTS0887) and Open-End Fund for the Valuable and Precision Instruments of Central South University (NO. CSUZC201837).