Role of microRNA-25 in high glucose cultured Müller glia
Autor: | Qinghuai Liu, Songtao Yuan, Han Shen, Yu Liu |
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Jazyk: | angličtina |
Rok vydání: | 2021 |
Předmět: |
medicine.diagnostic_test
business.industry proliferation Immunocytochemistry apoptosis müller glia Transfection RE1-994 Cell counting Molecular biology Flow cytometry microrna-25 Reverse transcription polymerase chain reaction Ophthalmology Basic Research medicine.anatomical_structure Apoptosis microRNA medicine business Muller glia |
Zdroj: | International Journal of Ophthalmology, Vol 14, Iss 5, Pp 643-648 (2021) Int J Ophthalmol |
ISSN: | 2227-4898 2222-3959 |
Popis: | 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. |
Databáze: | OpenAIRE |
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