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Mesenchymal stem cell-derived exosomal CBLB ameliorates infantile pneumonia progression probably by ubiquitinating
Fang Guo1, Fuxing Song1, Zhenjiang Chen1
1Department of Pediatrics, Jinan City People's Hospital, No. 001, Xuehu Street, Changshao North Road, Laiwu District, Jinan, 271100, China.
Background:
Infantile pneumonia (IP) is a significant cause of morbidity and mortality in young children. Mesenchymal stem cells (MSCs) have emerged as potential therapeutic agents in pneumonia due to their immunomodulatory properties. The study analyzed the role of MSCs from bone marrow in IP and the underlying mechanism.
Methods:
Human embryonic lung fibroblasts (WI-38) were stimulated using lipopolysaccharide (LPS) to mimic an IP cell model. This study employed flow cytometry to analyze the expression of hematopoietic markers and marker proteins on MSCs. The differentiation potential of MSCs was assessed through microscopy, oil red O staining, and alkaline phosphatase (ALP) assays. The localization of exosomes in WI-38 cells was observed using the cell membrane green fluorescent probe DIO. Quantitative reverse transcription polymerase chain reaction (qRT-PCR), western blotting and immunohistochemistry assays were used to analyze the expression of mRNA or protein. Cell viability, proliferation, and apoptosis were evaluated using Cell counting kit-8, 5-Ethynyl-2'-deoxyuridine, and flow cytometry assays, respectively. Enzyme-linked immunosorbent assays were conducted to measure cytokine levels. A mouse model of pneumonia was utilized to assess the therapeutic potential of MSC-derived exosomes on lung injury. Co-immunoprecipitation (Co-IP) assay was performed to study the interaction between Cbl proto-oncogene B (CBLB) and mitogen-activated protein kinase 14 (MAPK14).
Results:
MSC-derived exosomes could be transferred into LPS-induced WI-38 cells, where they mitigated the inhibitory effects of LPS on CBLB mRNA expression. These exosomes improved WI-38 cell proliferation, reduced apoptosis, and decreased the production of pro-inflammatory cytokines including IL-6, IL-1β, and TNF-α by regulating CBLB after LPS treatment. Moreover, in a mouse model, MSC-derived exosomes protected against LPS-induced lung injury, whereas the effect was reversed after treatment with the exosomes isolated from CBLB-deficient MSCs. In addition, CBLB was found to destabilize MAPK14 protein expression in WI-38 cells. Further, overexpression of CBLB ameliorated LPS-induced inhibitory effect on cell proliferation and promoting effects on cell apoptosis and inflammation in WI-38 cells by regulating MAPK14.
Conclusion:
MSC-derived exosomal CBLB has therapeutic potential in ameliorating the progression of IP probably by ubiquitinating MAPK14, which could lead to novel clinical interventions for treating this condition.
Insights
Mesenchymal stem cell-derived exosomes carrying CBLB show promise in treating infantile pneumonia (IP). These exosomes protect lung cells and reduce inflammation by targeting MAPK14, offering a potential new therapy for this childhood disease.
Area of Science:
- Biomedical research
- Cell biology
- Immunology
Background:
- Infantile pneumonia (IP) is a major cause of illness and death in children.
- Mesenchymal stem cells (MSCs) possess immunomodulatory capabilities, making them promising for pneumonia treatment.
Purpose of the Study:
- To investigate the therapeutic role of bone marrow-derived MSCs in infantile pneumonia.
- To elucidate the underlying molecular mechanisms involving exosomes and specific proteins.
Main Methods:
- Established a cell model of IP using lipopolysaccharide (LPS)-stimulated lung fibroblasts (WI-38).
- Analyzed MSCs using flow cytometry and assessed differentiation potential.
- Utilized qRT-PCR, western blotting, and immunohistochemistry to measure gene and protein expression.
- Evaluated cell viability, proliferation, and apoptosis, and measured cytokine levels.
- Assessed therapeutic effects in a mouse model of pneumonia and investigated protein interactions via co-immunoprecipitation.
Main Results:
- MSC-derived exosomes were internalized by LPS-treated WI-38 cells, mitigating LPS-induced suppression of CBLB mRNA.
- Exosomes enhanced cell proliferation, reduced apoptosis, and decreased pro-inflammatory cytokines (IL-6, IL-1β, TNF-α) by regulating CBLB.
- In vivo, MSC-derived exosomes protected against LPS-induced lung injury, with effects reversed by CBLB-deficient exosomes.
- CBLB destabilizes MAPK14 protein expression; CBLB overexpression counteracted LPS-induced negative effects on proliferation and inflammation via MAPK14 regulation.
Conclusions:
- MSC-derived exosomal CBLB demonstrates therapeutic potential for infantile pneumonia.
- The mechanism involves ubiquitinating MAPK14, suggesting a novel pathway for clinical intervention.
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