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DEPTOR alleviates LPS-induced inflammation and ER stress in WI-38 cells
Xiangxiang Shi1, Jin Ding2, Bihe Zeng3
1Department of Pediatric Internal Medicine, Huai'an Second People's Hospital, the Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an City, Jiangsu Province, China.
Insights
DEP domain-containing mTOR-interacting protein (DEPTOR) protects lung cells from lipopolysaccharide (LPS)-induced damage. Overexpressing DEPTOR reduces inflammation, oxidative stress, and apoptosis, suggesting its therapeutic potential for pneumonia.
Area of Science:
- Cellular Biology
- Molecular Medicine
- Immunology
Background:
- Pediatric pneumonia involves inflammation, oxidative stress, and apoptosis, often triggered by lipopolysaccharide (LPS).
- DEP domain-containing mTOR-interacting protein (DEPTOR) regulates inflammation and ER homeostasis but its role in pneumonia is unclear.
Purpose of the Study:
- To investigate the role of DEPTOR in lipopolysaccharide (LPS)-induced lung cell injury.
- To determine if DEPTOR overexpression can mitigate LPS-induced cellular damage.
Main Methods:
- An in vitro model using WI-38 human fetal lung fibroblast cells stimulated with LPS.
- DEPTOR was overexpressed using plasmid transfection.
- Assessed pro-inflammatory cytokine release, oxidative/ER stress, apoptosis, and NF-κB signaling via qPCR, Western blot, ELISA, flow cytometry, and biochemical assays.
Main Results:
- DEPTOR expression decreased in LPS-stimulated cells.
- DEPTOR overexpression suppressed LPS-induced cytokine production (P < 0.01).
- DEPTOR overexpression reduced oxidative and ER stress markers and inhibited apoptosis (P < 0.01).
Conclusions:
- DEPTOR exhibits a protective effect against LPS-induced cellular injury.
- DEPTOR may serve as a potential therapeutic target for mitigating pneumonia.
Background:
Pediatric pneumonia is a severe inflammatory condition frequently precipitated by bacterial endotoxins, such as lipopolysaccharide (LPS), which can elicit oxidative stress, endoplasmic reticulum (ER) stress, and apoptotic cell death. DEP domain-containing mTOR-interacting protein (DEPTOR), an endogenous inhibitor of mTOR signaling, has been implicated in the regulation of inflammation and ER homeostasis. However, its specific function in the pathogenesis of pneumonia remains poorly defined.
Methods:
WI-38 human fetal lung fibroblast cells were employed to establish an in vitro model of LPS-induced inflammation. DEPTOR was overexpressed via plasmid transfection to examine its functional role. The impact of DEPTOR on pro-inflammatory cytokine release, oxidative and ER stress responses, apoptosis, and nuclear factor kappa B signaling was comprehensively evaluated using quantitative real-time polymerase chain reaction, Western blot analysis, enzyme-linked-immunosorbent serologic assay, flow cytometry, and biochemical assays.
Results:
DEPTOR expression is significantly downregulated in LPS-stimulated WI-38 cells (P < 0.01). DEPTOR overexpression markedly suppresses LPS-induced pro-inflammatory cytokine production (P < 0.01), ameliorates oxidative and ER stress-as indicated by decreased lipid peroxidation and restoration of superoxide dismutase and glutathione levels (P < 0.01)-and inhibits apoptosis, reducing apoptotic cell percentages by over 10% (P < 0.01).
Conclusion:
These results suggest that DEPTOR confers a protective role against LPS-induced cellular injury, supporting its potential as a promising therapeutic target for mitigating.
