于丰萁 1马汀 1赵伟伟 1杜文迪 1姜镁钧 1胡荣荣1
作者信息
- 1. 266071 山东省青岛市,中国人民解放军海军第九七一医院眼科
- 折叠
摘要
Abstract
Objective To explore the effects of graphene quantum dots(GQDs)on the reprogramming of Müller cells.Methods Müller cells were randomly divided into the control,dedifferentiation,dedifferentiation+GQD,and GQD groups.The cells in the dedifferentiation group and the dedifferentiation+GQD group were dedifferentiated using the serum-free medium containing DMEM/F12(1∶1),20 μg·L-1 EGF,10 μg·L-1 bFGF,2 × B27,and 1 × N2 for 5 d.Then,the cells in the dedifferentiation+GQD group and the GQD group were treated with 50 mg·L-1 GQDs for 72 h.The Müller cells in the control group were subjected to normal cell culture.Immunofluorescence staining was used to identify the ex-pression of Müller cell markers,including glial fibrillary acidic protein(GFAP),glutamate-aspartate transporter(GLAST),and glutamine synthetase(GS).Phalloidin staining was performed to observe whether Müller cells could take up GQDs.The effect of different concentrations of GQDs on the proliferation of Müller cells was assessed using the cell counting kit-8(CCK-8)assay.The effect of GQDs on the expression of neural progenitor/stem cell markers(SRY-box transcription factor 2[SOX-2]and Nestin)and the astrocyte marker GFAP in normal and dedifferentiated Müller cells was examined using im-munofluorescence staining and Western blotting.Results The immunofluorescence staining results showed that the fluo-rescence of GFAP was extremely weak and almost invisible in Müller cells,while the fluorescence of GLAST and GS was extremely strong and predominantly appeared in the cytoplasm of Müller cells.After 24 h of GQD treatment,trace amounts of GQD fluorescence were visible in Müller cells.The amount of GQD fluorescence in the cytoplasm of Müller cells gradual-ly increased with time.The CCK-8 assay results showed that the activity of Müller cells tended to decrease with an increase in the treatment time and concentration of GQD.The statistical analysis results showed that the mean fluorescence intensity of GFAP,Nestin,and SOX-2 in Müller cells of the dedifferentiation,dedifferentiation+GQD,and GQD groups was higher than that of the control group,and the differences were statistically significant(all P＜0.05).The mean fluorescence inten-sity of GFAP in the dedifferentiation+GQD group was higher than that in the dedifferentiation group,the mean fluores-cence intensity of Nestin and SOX-2 was lower than that in the dedifferentiation group,and the differences were all statisti-cally significant(P＜0.05).The relative protein expression of GFAP,Nestin,and SOX-2 in Müller cells in the dedifferentia-tion,dedifferentiation+GQD,and GQD groups was higher than that in the control group,and the differences were statis-tically significant(all P＜0.05).The relative protein expression of GFAP in the dedifferentiation+GQD group was higher than that in the dedifferentiation group,the relative protein expression of Nestin and SOX-2 was lower than that in the dedi-fferentiation group,and the differences were statistically significant(all P＜0.05).Conclusion GQDs facilitate the re-programming of Müller cells into astrocytes.关键词
Müller细胞/星形胶质细胞/石墨烯量子点/分化Key words
Müller cell/astrocyte/graphene/differentiation分类
临床医学
