The concentration-dependent pro-fibrotic effect of metformin on LPS and high glucose induced fibroblast NIH 3T3 and
R Awaluddin1, D A A Nugrahaningsih2, E N Solikhah3
1Universitas Gadjah Mada, Public Health, and Nursing, Faculty of Medicine, Master in Biomedical Science Program, Jl. Farmako Sekip Utara, Yogyakarta, Indonesia.
Introduction:
Diabetes mellitus is known as one of the risk factors for Idiopathic Pulmonary Fibrosis (IPF) development. Recently, metformin, the commonly used antidiabetic medication, is reported to have a therapeutic effect in IPF. However, the benefit of metformin therapy in IPF is still controversial. The study aims to investigate the metformin effect on the fibroblast and macrophage co-culture under lipopolysaccharides (LPS) and high glucose treatment.
Method:
The NIH 3T3 and RAW 264.7 co-culture were induced with LPS and high glucose before it was treated with metformin in different concentration. After 24 hours of treatment, the media and the cells were collected for further examination. The collagen expression was measured using Sirius red dye in the media. The IL-6 and TGF β mRNA examination were done using real-time PCR.
Result:
Our study showed that NIH 3T3 and RAW 264.7 coculture treated with metformin has higher collagen expression, but lower IL-6 mRNA expression compares to those on co-culture without treatment.
Conclusion:
Metformin increases fibrosis markers in LPS and high glucose-induced NIH 3T3 and RAW 264.7 coculture despite its ability to improve IL-6 mRNA expression.
Insights
Metformin treatment in a cell model of Idiopathic Pulmonary Fibrosis (IPF) increased fibrosis markers, specifically collagen, while decreasing IL-6. This suggests metformin may worsen fibrosis despite anti-inflammatory effects in IPF.
Area of Science:
- Pulmonary Medicine
- Cell Biology
- Pharmacology
Background:
- Diabetes mellitus is a known risk factor for Idiopathic Pulmonary Fibrosis (IPF).
- Metformin, an antidiabetic drug, has shown potential therapeutic effects in IPF, but its benefits remain controversial.
- Investigating metformin's impact on IPF pathogenesis is crucial for understanding its role in disease progression.
Purpose of the Study:
- To investigate the effect of metformin on fibroblast and macrophage co-cultures.
- To examine metformin's influence under conditions mimicking IPF, including lipopolysaccharides (LPS) and high glucose.
- To clarify the controversial role of metformin in IPF treatment.
Main Methods:
- Co-culture of NIH 3T3 fibroblasts and RAW 264.7 macrophages.
- Induction of co-cultures with LPS and high glucose, followed by metformin treatment.
- Measurement of collagen expression using Sirius red dye and IL-6 and TGF-β mRNA levels via real-time PCR.
Main Results:
- Metformin treatment led to increased collagen expression in the co-culture.
- A decrease in IL-6 mRNA expression was observed with metformin treatment.
- The study utilized a co-culture model to assess metformin's impact on fibrosis and inflammation markers.
Conclusions:
- Metformin increased fibrosis markers (collagen) in the induced co-culture model.
- Metformin reduced IL-6 mRNA expression, suggesting an anti-inflammatory effect.
- The findings indicate that metformin may exacerbate fibrosis in IPF, despite improving certain inflammatory markers.
