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Mehrdad Afarid1, Hossein Bahari1, Fatemeh Sanie-Jahromi1
1Poostchi Ophthalmology Research Center, Department of Ophthalmology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Abstract:
Angiogenesis, retinal neuropathy, and inflammation are the main molecular features of diabetic retinopathy (DR) and should be taken into consideration for potential treatment approaches. Retinal pigmented epithelial (RPE) cells play a major role in DR progression. This study evaluated the in vitro effect of interferon (IFN) α-2b on the expression of genes involved in apoptosis, inflammation, neuroprotection, and angiogenesis in RPE cells. RPE cells were cocultured with IFN α-2b at 2 doses (500 and 1,000 IU) and treatment periods (24 and 48 h). The quantitative relative expression of genes (BCL-2, BAX, BDNF, VEGF, and IL-1b) was evaluated in the treated versus control cells through real-time polymerase chain reaction (PCR). The result of this study demonstrated that IFN treatment at 1,000 IU (48 h) led to significant upregulation of BCL-2, BAX, BDNF, and IL-1b; however, the BCL-2/BAX ratio was not statistically altered from 1:1, in any of the treatment patterns. We also showed that VEGF expression was downregulated in RPE cells treated with 500 IU for 24 h. It can be concluded that IFN α-2b was safe (BCL-2/BAX ∼1:1) and enhanced neuroprotection at 1,000 IU (48 h); however-at the same time-IFN α-2b induced inflammation in RPE cells. Moreover, the antiangiogenic effect of IFN α-2b was solely observed in RPE cells treated with 500 IU (24 h). It seems that IFN α-2b in lower doses and short duration exerts antiangiogenic effects and in higher doses and longer duration has neuroprotective and inflammatory effects. Hence, appropriate concentration and duration of treatment, according to the type and stage of the disease, should be considered to achieve success in IFN therapy.
Insights
Interferon (IFN) α-2b shows dual effects in diabetic retinopathy (DR) models. Lower doses reduce blood vessel growth, while higher doses offer neuroprotection but increase inflammation, requiring careful treatment selection.
Area of Science:
- Ophthalmology and Molecular Biology
- Diabetic Retinopathy (DR) Pathogenesis
- Retinal Pigmented Epithelial (RPE) Cell Biology
Background:
- Diabetic retinopathy (DR) is characterized by angiogenesis, retinal neuropathy, and inflammation.
- Retinal pigmented epithelial (RPE) cells are critical in DR progression.
- Understanding molecular mechanisms is key for developing effective DR treatments.
Purpose of the Study:
- To investigate the in vitro effects of interferon (IFN) α-2b on RPE cells.
- To analyze gene expression related to apoptosis, inflammation, neuroprotection, and angiogenesis.
- To determine optimal IFN α-2b dosage and duration for potential therapeutic applications in DR.
Main Methods:
- RPE cells were co-cultured with IFN α-2b at doses of 500 and 1,000 IU.
- Treatment durations included 24 and 48 hours.
- Quantitative real-time PCR was used to measure the expression of BCL-2, BAX, BDNF, VEGF, and IL-1b genes.
Main Results:
- IFN α-2b at 1,000 IU for 48 hours upregulated BCL-2, BAX, BDNF, and IL-1b, with a stable BCL-2/BAX ratio (~1:1).
- VEGF expression was downregulated in RPE cells treated with 500 IU for 24 hours, indicating an anti-angiogenic effect.
- Higher doses/longer duration promoted neuroprotection and inflammation, while lower doses/shorter duration showed anti-angiogenic effects.
Conclusions:
- IFN α-2b demonstrates dose- and time-dependent effects on RPE cells.
- It offers potential neuroprotection and anti-angiogenesis but also induces inflammation.
- Tailoring IFN α-2b concentration and treatment duration is crucial for managing DR based on disease stage and type.