Targeting TFE3 Protects Against Lysosomal Malfunction-Induced Pyroptosis in Random Skin Flaps via ROS Elimination

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.

Baicalin promotes random‐pattern skin flap survival by inducing autophagy via AMPK‐regulated TFEB nuclear transcription

“…Therefore, whether these kinases will be included in the therapy effect of BA needs further study. Third, in term of the theory of flap survival (Wu et al, 2019), the greater the chance of necrosis of the distal flap, the control group in this experimental model had complete necrosis in the Area III and a small part of survival in the Area II, which was consistent with the previous study (Li et al, 2021; Lin et al, 2018). Though BA treatment significantly improved the survival rate in Area II, it would be better to have a positive control group in this study.…”

“…When the flap size is too large or the flap's width‐to‐length ratio exceeds 1:2, narcosis develops in the distal portion of the flap (El Shaer et al, 2019). Therefore, a lot of studies have engaged to improve the survival of the skin flap including delayed surgery (Byun et al, 2020), biomaterial (Mao et al, 2019), and even gene intervention (Li et al, 2021). Despite these significant advancements, the therapeutic necessity to be able to harvest bigger portions of skin flaps remains unmet.…”

Development and Validation of a Pyroptosis-Related Long Non-coding RNA Signature for Hepatocellular Carcinoma

“…LncRNA Neat1 is associated with NLRP3 , NLRC4 , and AIM2 inflammasomes in mouse macrophages to increase their assembly and subsequent pro-caspase-1 processing ( Zhang et al, 2019 ). ROS-induced lysosome malfunction is a vital inducement of pyroptosis, and the arginine metabolism pathway participates in NLRP3 inflammasome inhibition ( Jiang et al, 2021 ; Li et al, 2021 ). Glycolytic metabolism can impose stress on DNA replication and activate AIM2 inflammasome ( Fidler et al, 2021 ).…”

Procyanidin B2 regulates the Sirt1/Nrf2 signaling pathway to improve random‐pattern skin flap survival

“…It has been reported that the accumulation of ROS results in neutrophil infiltration, increased protease release, and the formation of several oxidative intermediates (Sies & Jones, 2020). Excessive ROS generation causes partial or complete necrosis of the flap (Li et al, 2021). The first-line defense against accumulated superoxide anion radicals is SODs, while GSH-Px is upregulated to eliminate peroxides and hydroxyl radicals, and PB2's antioxidant effects were investigated using these enzymes (Taysi et al, 2019).…”