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Pirfenidone Inhibits Inflammatory Mediators in Human Cornea In Vitro
Alexandria C Hofmann1,2, Nishant R Sinha1,3,2, Abigail Hurtado3
1Harry S. Truman Memorial Veterans' Hospital, Columbia, Missouri, USA.
Purpose:
Corneal inflammation is a common clinical condition that contributes to loss of corneal transparency. This study evaluated the potential of pirfenidone (PFD), an FDA-approved drug, to inhibit inflammation in the human cornea using an in vitro model.
Methods:
Healthy cadaver human corneas and standard in vitro corneal inflammation models were used. Primary human corneal stromal fibroblasts (hCSFs) generated from donor corneas were used in the entire study. The dose-dependent assays identified optimal lipopolysaccharide (LPS; 0-150 ng/ml) concentration to induce inflammation and PFD's (0-500 µg/mL) half-maximal inhibitory concentration (IC50) to hCSF. The quantitative reverse transcriptase polymerase chain reaction, immunofluorescence, Western blotting, and reactive oxygen species (ROS) commercial kits studied changes in inflammatory response.
Results:
A significant upregulation of inflammatory markers in hCSF at 48 h was noted at 50 ng/ml of LPS concentration. The IC50 of PFD was found to be 200 µg/mL for hCSFs. Topical treatments of PFD (200 µg/mL) significantly reduced LPS (50 µg/mL)-induced levels of pro-inflammatory (TNFα, IL1α, IL1β, IL6, and cyclooxygenase 2) markers and oxidative stress (glutathione peroxidase 1 [GPX1], GPX4, and catalase) genes (P < 0.001 or P < 0.0001) in human corneas in vitro. Moreover, PFD significantly inhibited LPS-increased ROS production (P < 0.001) in hCSF. PFD treatment reduced LPS-induced inflammatory stimulation and tumor necrosis factor alpha and IκB alpha phosphorylation in hCSF.
Conclusion:
PFD inhibits corneal inflammation in vitro by reducing pro-inflammatory cytokines and oxidative stress. Comprehensive preclinical animal testing is warranted to define the potential of PFD to treat corneal symptoms in vivo in traumatic eyes.
Insights
Pirfenidone (PFD) effectively reduces corneal inflammation and oxidative stress in human corneal cells in vitro. Further animal studies are needed to explore PFD
Area of Science:
- Ophthalmology
- Cell Biology
- Pharmacology
Background:
- Corneal inflammation can impair vision by reducing corneal transparency.
- Pirfenidone (PFD) is an FDA-approved drug with anti-inflammatory properties.
Purpose of the Study:
- To evaluate the efficacy of pirfenidone (PFD) in inhibiting corneal inflammation using an in vitro model.
- To investigate PFD's effects on inflammatory markers and oxidative stress in human corneal cells.
Main Methods:
- Human corneal stromal fibroblasts (hCSFs) were used to establish an in vitro corneal inflammation model.
- Lipopolysaccharide (LPS) induced inflammation, and PFD's anti-inflammatory effects were assessed using molecular and cellular assays.
- Key inflammatory markers, oxidative stress indicators, and reactive oxygen species (ROS) production were quantified.
Main Results:
- LPS significantly upregulated inflammatory markers and ROS production in hCSFs.
- PFD treatment (200 µg/mL) significantly reduced LPS-induced pro-inflammatory cytokines (TNFα, IL1α, IL1β, IL6, COX-2) and oxidative stress markers (GPX1, GPX4, catalase).
- PFD inhibited LPS-induced ROS production and phosphorylation of key inflammatory signaling molecules.
Conclusions:
- PFD demonstrates significant anti-inflammatory and anti-oxidative stress effects in human corneal cells in vitro.
- These findings suggest PFD's potential therapeutic value for corneal inflammatory conditions.
- Further preclinical in vivo studies are recommended to validate PFD's efficacy in treating corneal inflammation.
