GMFB在氧化应激诱导的ARPE19细胞中的表达及意义

目的 探讨胶质细胞成熟因子beta(GMFB)在氧化应激诱导的ARPE19细胞中的表达及意义。方法 用0、0.25、0.5、1、2mmol乙二醛处理ARPE19细胞24h,其中对照组用0mmol/L乙二醛处理,实验组以0.25～2mmol/L乙二醛处理,用Western印迹法检测GMFB的表达；实验组用1μg/ml GMFB处理ARPE19细胞24h,对照组不做处理,进行RNAseq,比较实验组和对照组基因表达的差异,选取其中表达差异大于1.5倍且P＜0.05的基因作为差异表达的基因,对其进行GO及KEGG富集分析。结果 经乙二醛处理的各组ARPE19细胞GMFB表达上调,以0.5mmol/L处理组的GMFB上调最为显著。对GMFB处理的ARPE19细胞进行全基因表达谱检测,结果显示,在被检测的19966个基因中,实验组和正常组间有显著差异表达(改变1.5倍)的基因有583个。与对照组相比,实验组基因表达显著上调的有265个,下调的有318个。GO分析表明,GMFB与细胞代谢、细胞催化等生物过程相关。KEGG分析显示,GMFB与细胞代谢通路、癌症通路、Wnt信号通路、趋化因子信号通路等多条信号通路密切相关。炎症基因CCL26、CXCL8和NF-κB1表达升高,提示GMFB与炎症相关。结论 GMFB在氧化应激诱导的ARPE19细胞中表达明显上调,GMFB可能影响细胞间连接,参与Wnt信号通路、肿瘤相关信号通路和趋化因子信号通路等。
Objective To investigate the expression of glia maturation factor beta(GMFB) in human retinal pigment epithelium(RPE) ARPE19 cells and its relation to oxidative stress. Methods ARPE19 cells were treated with glyoxal(0, 0.25, 0.5, 1.0 and 2.0mmol/L) for 24h. GMFB expression levels were examined by Western blot. APPE19 was treated with 1μg/ml GMFB for 24h. Total RNA was extracted and used for RNAseq assay. By comparing the differentially expressed profile of two groups, the genes with expression difference fold ＞1.5 times and P＜0.05 were selected as candidate genes for GO enrichment analysis and KEGG analysis. Results In the ARPE19 cells under the treatments of different concentrations of glyoxal, GMFB expression levels were significantly up-regulated. RNAseq analysis showed that the expression patterns of 583 genes significantly altered in experimental group, of which 265 were up-regulated and 318 were down-regulated. GO analysis indicated that GMFB was involved in the process of cellular metabolism. KEGG analysis further indicated that GMFB was involved in metabolic pathway, cancer pathway, Wnt signaling pathway and chemokine signaling pathway. Furthermore, inflammation related genes CCL26, CXCL8 and NFKB1 were also up-regulated. Conclusion Oxidative stress induces the up-regulation of GMFB in ARPE19 cells. The effects of GMFB on ARPE19 were related with cell-cell binding, Wnt signaling pathway, pathway in cancer as well as chemokine signaling pathway. The results indicate that GMFB is involved in important cellular process in RPE cells under oxidative stress and might be an important factor in the pathology of retinal degeneration