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...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-q PCR).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.RTPCR results showed that the expression level of miR-25 was significantly repressed in HGMG,while overexpression 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.展开更多
基金Supported by National Key Research and Development Program of China(No.2017YFA0104101)。
文摘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-q PCR).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.RTPCR results showed that the expression level of miR-25 was significantly repressed in HGMG,while overexpression 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.
