石墨烯量子点对Müller细胞重编程的影响  

Effects of graphene quantum dots on the reprogramming of Müller cells

作　　者：于丰萁 马汀 赵伟伟 杜文迪 姜镁钧 胡荣荣 YU Fengqi;MA Ting;ZHAO Weiwei;DU Wendi;JIANG Meijun;HU Rongrong(Department of Ophthalmology,the 971 Naval Hospital of PLA,Qingdao 266071,Shandong Province,China)

机构地区：[1]中国人民解放军海军第九七一医院眼科,山东省青岛市266071

出　　处：《眼科新进展》2025年第5期354-358,共5页Recent Advances in Ophthalmology

摘　　要：目的探讨石墨烯量子点(graphene quantum dots,GQD)对Müller细胞重编程的影响。方法取Müller细胞随机分为对照组、去分化组、去分化+GQD组、及GQD组。去分化组和去分化+GQD组细胞由含DMEM/F12(1 GA6FA 1)、20μg·L^(-1)EGF、10μg·L^(-1)bFGF、2×B27和1×N2组成的无血清培养基去分化5 d;之后将去分化+GQD组及GQD组细胞采用50 mg·L^(-1)GQD处理72 h。对照组Müller细胞仅进行正常的细胞培养。免疫荧光染色鉴定Müller细胞标志物神经胶质纤维酸性蛋白(GFAP)、谷氨酸-天冬氨酸转运蛋白(GLAST)和谷氨酰胺合成酶(GS)的表达。鬼笔环肽染色观察Müller细胞是否能够摄取GQD。CCK-8实验评估不同浓度GQD对Müller细胞增殖的影响。免疫荧光染色和免疫印迹法检测GQD对正常和去分化的Müller细胞中神经祖/干细胞标志物SRY-Box转录因子2(SOX-2)和巢蛋白(Nestin)及星形胶质细胞标志物GFAP表达的影响。结果免疫荧光染色结果显示,Müller细胞中GFAP的荧光极弱,几乎不可见;GLAST和GS的荧光极强,并且主要出现在细胞质中。GQD处理24 h后,Müller细胞的胞浆中可见微量的GQD荧光。随着处理后时间的延长,Müller细胞的胞浆中GQD荧光数量逐渐增加。CCK-8实验结果显示,随着GQD处理时间的延长以及浓度增大,Müller细胞活性呈现降低的趋势。统计结果分析显示,去分化组、去分化+GQD组和GQD组Müller细胞中GFAP、Nestin和SOX-2的平均荧光强度均高于对照组,差异均有统计学意义(均为P<0.05);去分化+GQD组Müller细胞中GFAP的平均荧光强度高于去分化组,且Nestin和SOX-2的平均荧光强度均低于去分化组,差异均有统计学意义(均为P<0.05)。去分化组、去分化+GQD组和GQD组Müller细胞中GFAP、Nestin和SOX-2的蛋白相对表达量均高于对照组,差异均有统计学意义(均为P<0.05);去分化组+GQD组Müller细胞中GFAP的蛋白相对表达量高于去分化组,且Nestin和SOX-2的蛋白相对表达�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 GA6FA 1),20μg·L^(-1)EGF,10μg·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 expression 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 immunofluorescence staining and Western blotting.Results The immunofluorescence staining results showed that the fluorescence 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 gradually 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 d

关 键 词：MÜLLER细胞 星形胶质细胞 石墨烯量子点 分化 

分 类 号：R774[医药卫生—眼科]