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MCC950 inhibits the inflammatory response and excessive proliferation of canine corneal stromal cells induced by
Long Guo1, Zhihao Wang1, Chengcheng Zhu1
1College of Veterinary Medicine, Yangzhou University, Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu 225009, China; Joint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of Education, Yangzhou, Jiangsu 225009, China; International Research Laboratory of Prevention and Control of Important Animal Infectious Diseases and Zoonotic Diseases of Jiangsu Higher Education Institutions, Yangzhou University, Yangzhou, China.
Background:
The stromal layer is the thickest layer of the cornea, and corneal stromal cells play an important role in the inflammatory response and wound repair. This study investigated the effect of MCC950, an inhibitor of NLRP3 inflammasome, on the inflammatory response and proliferation of canine corneal stromal cells (CCSCs) induced by Staphylococcus pseudintermedius (S. pseudintermedius).
Methods:
CCSCs were pretreated with MCC950 and infected with S. pseudintermedius. The phosphorylation of p65, IκBα, PI3K, and AKT and the expression of NLRP3, caspase-1 p20, cleaved IL-1β, ASC, β-catenin, c-Myc, and CyclinD1 were detected by western blotting. The expression of inflammatory factors (IL-1β, IL-6, IL-8, IL-18, and TNF-α) and growth factors (EGF, FGF, TGF-β1, VEGF, and CTGF) were measured by RT-PCR. The levels of MDA content and LDH activity were detected by an assay kit. The cell cycle was detected by flow cytometry.
Results:
MCC950 down-regulated the phosphorylation of p65, IκBα, PI3K, and AKT and decreased the expression of NLRP3, caspase-1 p20, cleaved IL-1β, ASC, β-catenin, c-Myc, and CyclinD1 compared to those in the S. pseudintermedius infection group (p < 0.05). MCC950 significantly inhibited the expression of inflammatory factors (IL-1β, IL-6, IL-8, IL-18, and TNF-α) and growth factors (EGF, FGF, TGF-β1, VEGF, and CTGF) induced by S. pseudintermedius (p < 0.01). Compared to the S. pseudintermedius infection group, the MDA content and LDH activity of CCSCs were significantly decreased after treatment with MCC950 (p < 0.01).
Conclusion:
MCC950 attenuates S. pseudintermedius-induced inflammatory responses in CCSCs. At the same time, MCC950 can inhibit excessive proliferation of cells, which is beneficial for alleviating corneal fibrosis healing.
Insights
MCC950, an NLRP3 inflammasome inhibitor, reduces Staphylococcus pseudintermedius-induced inflammation and excessive cell proliferation in canine corneal stromal cells (CCSCs). This finding suggests MCC950 may help alleviate corneal fibrosis during healing.
Area of Science:
- Veterinary Ophthalmology
- Immunology
- Cell Biology
Background:
- Corneal stromal cells are crucial for immune response and wound healing.
- Staphylococcus pseudintermedius infection triggers inflammation and affects corneal cell function.
Purpose of the Study:
- To investigate the effects of MCC950, an NLRP3 inflammasome inhibitor, on canine corneal stromal cells (CCSCs) in response to Staphylococcus pseudintermedius infection.
- To evaluate MCC950's impact on inflammatory pathways and cell proliferation.
Main Methods:
- CCSCs were pretreated with MCC950 and infected with S. pseudintermedius.
- Western blotting, RT-PCR, and flow cytometry were used to analyze protein and gene expression, inflammatory markers, and cell cycle.
- MDA content and LDH activity were measured to assess cell damage.
Main Results:
- MCC950 significantly reduced the phosphorylation of key inflammatory signaling molecules (p65, IκBα, PI3K, AKT).
- MCC950 decreased the expression of NLRP3 inflammasome components and inflammatory cytokines (IL-1β, IL-6, IL-8, IL-18, TNF-α).
- MCC950 inhibited the expression of growth factors and reduced cell proliferation and damage markers (MDA, LDH).
Conclusions:
- MCC950 effectively attenuates S. pseudintermedius-induced inflammatory responses in CCSCs.
- MCC950 inhibits excessive cell proliferation, which is beneficial for managing corneal fibrosis and promoting healing.
