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Published on: January 3, 2025
Observation of inflammation-induced mitophagy during stroke by a mitochondria-targeting two-photon ratiometric probe
Fei Cheng1, Taotao Qiang2, Longfang Ren1
1College of Bioresources and Materials Engineering, Shaanxi University of Science & Technology, Xi'an, 710021, China. qiangtt515@163.com huwchem@163.com.
Abstract:
This study reports the development of a new, pH-sensitive, mitochondria-targeting two-photon ratiometric probe (Mito-BNO) for real-time tracking of mitophagy, a process that can be accelerated in brain tissue during stroke. Mito-BNO shows excellent capability for mitochondrial localisation (Pearson's correlation coefficient, r = 0.91), and can also effectively distinguish mitochondria from other subcellular organelles such as lysosomes and the endoplasmic reticulum (r = 0.40 and r = 0.33, respectively). Meanwhile, a rewarding pKa value (5.23 ± 0.03) and the pH reversibility suggest that Mito-BNO can track mitophagy in real time via confocal imaging. Most importantly, the relationship between mitophagy and neuroinflammation during stroke has been successfully demonstrated by evaluating the fluorescence of PC12 cells stained with Mito-BNO during an oxygen-glucose deprivation/reperfusion (OGD/R) process with and without anti-inflammatory treatment. The results indicate that the occurrence of mitophagy during stroke is caused by oxidative stress induced by neuroinflammation. This study will help further understanding stroke pathogenesis, can provide potential new targets for early diagnosis and treatment, and can also help to develop therapeutic drugs for stroke.
Insights
Researchers developed Mito-BNO, a pH-sensitive probe, to track mitophagy during stroke. This new tool reveals mitophagy is triggered by neuroinflammation-induced oxidative stress, offering insights into stroke pathogenesis.
Area of Science:
- Biomedical Engineering
- Cell Biology
- Neuroscience
Background:
- Mitophagy, the selective degradation of mitochondria, plays a critical role in cellular homeostasis.
- Accelerated mitophagy is observed in brain tissue during stroke, a condition characterized by neuroinflammation and oxidative stress.
- Understanding the real-time dynamics of mitophagy during stroke is crucial for developing effective therapeutic strategies.
Purpose of the Study:
- To develop a novel pH-sensitive, mitochondria-targeting two-photon ratiometric probe (Mito-BNO) for real-time mitophagy tracking.
- To investigate the relationship between mitophagy and neuroinflammation in the context of stroke.
- To elucidate the role of oxidative stress in mitophagy during stroke pathogenesis.
Main Methods:
- Synthesis and characterization of the Mito-BNO probe.
- Evaluation of Mito-BNO's specificity for mitochondria and its pH sensitivity using confocal imaging.
- Application of Mito-BNO in PC12 cells undergoing oxygen-glucose deprivation/reperfusion (OGD/R) to model stroke conditions.
- Assessment of mitophagy levels with and without anti-inflammatory treatment during OGD/R.
Main Results:
- Mito-BNO demonstrated excellent mitochondrial localization (r = 0.91) and specificity over other organelles.
- The probe exhibited a suitable pKa (5.23 ± 0.03) and pH reversibility, enabling real-time mitophagy tracking.
- The study successfully demonstrated that mitophagy during stroke is induced by oxidative stress resulting from neuroinflammation.
- Anti-inflammatory treatment modulated mitophagy, confirming the link between neuroinflammation and mitophagy during stroke.
Conclusions:
- Mito-BNO is a valuable tool for real-time monitoring of mitophagy in cellular models of stroke.
- Neuroinflammation-induced oxidative stress is a key driver of mitophagy during stroke.
- These findings enhance the understanding of stroke pathogenesis and suggest potential therapeutic targets for stroke treatment.
