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Updated: Nov 5, 2025
Primary Cell Cultures to Study the Regeneration Potential of Murine Müller Glia after MicroRNA Treatment
Published on: March 28, 2022
Role of microRNA-25 in high glucose cultured Müller glia
Yu Liu1, Han Shen1, Song-Tao Yuan1
1Department of Ophthalmology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, Jiangsu Province, China.
Aim:
To investigate the role of microRNA-25 (miR-25) in proliferation and apoptosis of retinal Müller glia (MG) under high glucose condition.
Methods:
The purity of the cultured cells was verified by immunocytochemistry and flow cytometry using antibodies that specifically recognized MG. The expression level of miR-25 under normal and high glucose conditions were validated by quantitative reverse transcription polymerase chain reaction (RT-qPCR). miR-25 mimics and negative control were transfected into MG and multiple functional experiments including cell counting kit-8 assay, EDU assay, and flow cytometry were conducted to explore the effects of miR-25 on the proliferation and apoptosis of high glucose cultured MG (HGMG).
Results:
Immunocytochemistry and flow cytometry confirmed the high purity of primary cultured MG. RT-PCR results showed that the expression level of miR-25 was significantly repressed in HGMG, while over-expression of miR-25 by miR-25 mimic markedly inhibited the high glucose induced cell apoptosis and promoted the proliferation of MG.
Conclusion:
The expression level of miR-25 is significantly downregulated in HGMG and its overexpression could attenuate the high glucose damages on MG by promoting proliferation and reducing apoptosis.
Insights
MicroRNA-25 (miR-25) is downregulated in high glucose conditions, impacting retinal Müller glia (MG). Overexpressing miR-25 protects MG by promoting proliferation and reducing apoptosis.
Area of Science:
- Ophthalmology
- Molecular Biology
- Cell Biology
Background:
- Diabetic retinopathy is a leading cause of blindness.
- High glucose conditions induce cellular stress and damage in retinal Müller glia (MG).
- MicroRNAs (miRNAs) play crucial roles in regulating cellular processes.
Purpose of the Study:
- To investigate the role of microRNA-25 (miR-25) in the proliferation and apoptosis of retinal Müller glia (MG) under high glucose conditions.
- To determine if miR-25 expression is altered in MG exposed to high glucose.
- To explore the therapeutic potential of miR-25 in mitigating high glucose-induced damage to MG.
Main Methods:
- Primary retinal Müller glia (MG) were cultured and exposed to normal or high glucose conditions.
- Cell purity was confirmed using immunocytochemistry and flow cytometry.
- Quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to measure miR-25 expression.
- miR-25 mimics were transfected into MG to assess its effects on proliferation and apoptosis via cell counting kit-8, EDU assay, and flow cytometry.
Main Results:
- High purity of primary cultured MG was confirmed.
- miR-25 expression was significantly downregulated in MG under high glucose conditions (HGMG).
- Overexpression of miR-25 via mimics inhibited high glucose-induced apoptosis and promoted MG proliferation.
Conclusions:
- miR-25 expression is significantly downregulated in high glucose-induced Müller glia (HGMG).
- Overexpression of miR-25 attenuates high glucose-induced damage to MG.
- miR-25 promotes MG proliferation and reduces apoptosis, suggesting a protective role.
