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Targeting TFE3 Protects Against Lysosomal Malfunction-Induced Pyroptosis in Random Skin Flaps via ROS Elimination
Jiafeng Li1,2, Junsheng Lou1,2, Gaoxiang Yu1,2
1Department of Orthopaedics, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, China.
Abstract:
Increasing evidence indicates that pyroptosis, a new type of programmed cell death, may participate in random flap necrosis and play an important role. ROS-induced lysosome malfunction is an important inducement of pyroptosis. Transcription factor E3 (TFE3) exerts a decisive effect in oxidative metabolism and lysosomal homeostasis. We explored the effect of pyroptosis in random flap necrosis and discussed the effect of TFE3 in modulating pyroptosis. Histological analysis via hematoxylin-eosin staining, immunohistochemistry, general evaluation of flaps, evaluation of tissue edema, and laser Doppler blood flow were employed to determine the survival of the skin flaps. Western blotting, immunofluorescence, and enzyme-linked immunosorbent assays were used to calculate the expressions of pyroptosis, oxidative stress, lysosome function, and the AMPK-MCOLN1 signaling pathway. In cell experiments, HUVEC cells were utilized to ensure the relationship between TFE3, reactive oxygen species (ROS)-induced lysosome malfunction and cell pyroptosis. Our results indicate that pyroptosis exists in the random skin flap model and oxygen and glucose deprivation/reperfusion cell model. In addition, NLRP3-mediated pyroptosis leads to necrosis of the flaps. Moreover, we also found that ischemic flaps can augment the accumulation of ROS, thereby inducing lysosomal malfunction and finally initiating pyroptosis. Meanwhile, we observed that TFE3 levels are interrelated with ROS levels, and overexpression and low expression of TFE3 levels can, respectively, inhibit and promote ROS-induced lysosomal dysfunction and pyroptosis during in vivo and in vitro experiments. In conclusion, we found the activation of TFE3 in random flaps is partially regulated by the AMPK-MCOLN1 signal pathway. Taken together, TFE3 is a key regulator of ROS-induced pyroptosis in random skin flaps, and TFE3 may be a promising therapeutic target for improving random flap survival.
Insights
Pyroptosis, a programmed cell death, contributes to random flap necrosis. Transcription factor E3 (TFE3) regulates reactive oxygen species (ROS)-induced pyroptosis, offering a potential therapeutic target for flap survival.
Area of Science:
- Biomedical Engineering
- Cell Biology
- Surgical Research
Background:
- Pyroptosis, a novel programmed cell death, is increasingly implicated in random flap necrosis.
- Reactive oxygen species (ROS)-induced lysosomal malfunction is a key trigger for pyroptosis.
- Transcription factor E3 (TFE3) plays a critical role in regulating oxidative metabolism and lysosomal homeostasis.
Purpose of the Study:
- To investigate the role of pyroptosis in random flap necrosis.
- To explore how Transcription factor E3 (TFE3) modulates pyroptosis in this context.
- To identify potential therapeutic targets for enhancing random flap survival.
Main Methods:
- Histological analysis (H&E staining), flap evaluation, and laser Doppler blood flow measurement for in vivo assessment.
- Western blotting, immunofluorescence, and ELISA for quantifying pyroptosis, oxidative stress, lysosomal function, and signaling pathway components.
- In vitro experiments using HUVEC cells to elucidate the TFE3-ROS-lysosome-pyroptosis axis.
Main Results:
- Pyroptosis was confirmed in both random skin flap and oxygen/glucose deprivation/reperfusion cell models.
- NLRP3-mediated pyroptosis was identified as a cause of flap necrosis.
- Ischemia-induced ROS accumulation triggers lysosomal malfunction and pyroptosis, with TFE3 levels inversely correlating with ROS and pyroptosis severity.
- TFE3 activation in flaps is partly regulated by the AMPK-MCOLN1 pathway.
Conclusions:
- Transcription factor E3 (TFE3) is a key regulator of ROS-induced pyroptosis in random skin flaps.
- TFE3 modulates pyroptosis by influencing ROS levels and lysosomal function.
- TFE3 represents a promising therapeutic target for improving random flap survival.
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