论文部分内容阅读
目的探讨HXO-RB44细胞上清液对人视网膜色素上皮(RPE)细胞系ARPE-19细胞内血管内皮生长因子(VEGF)表达的影响及其相关机制。方法分别利用含有10%胎牛血清的RPMI-1640培养基及MEM培养基对HXO-RB44细胞及ARPE-19细胞进行培养,在ARPE-19细胞内加入100μL HXO-RB44细胞上清液,分别干预0、4、8、12、24 h,利用RT-PCR、免疫荧光细胞化学、Western blot等方法,检测不同浓度的MLT干预后24 h ARPE-19细胞内VEGF mRNA及蛋白的表达情况。结果HXO-RB44细胞上清液加入后的不同时间点,VEGF mRNA的表达差异有统计学意义(F=195.072,P=0.000)。随着HXO-RB44细胞上清液作用时间的延长,ARPE-19细胞内VEGF mRNA表达逐渐增高(P<0.01)。HXO-RB44细胞上清液加入后4、8、12、24 h,ARPE-19细胞内VEGF蛋白表达显著增加(F=160.687,P=0.000),其表达与ARPE-19细胞内VEGF mRNA表达方式相同(P<0.05)。随着HXO-RB44细胞上清液作用时间的延长,ARPE-19细胞内绿荧光蛋白强度逐渐增加。结论HXO-RB44细胞上清液能促进ARPE-19细胞内VEGF的表达。
Objective To investigate the effect of HXO-RB44 cell supernatant on the expression of vascular endothelial growth factor (VEGF) in human retinal pigment epithelium (RPE) cell line ARPE-19 and its related mechanisms. Methods HXO-RB44 cells and ARPE-19 cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum and MEM medium respectively. Supernatants of HXO-RB44 cells (100 μL) were added to ARPE-19 cells, The expression of VEGF mRNA and protein in ARPE-19 cells were detected by RT-PCR, immunofluorescence cytochemistry, Western blot and other methods at different concentrations of MLT for 24 h. Results The expression of VEGF mRNA in HXO-RB44 cells at different time points was significantly different (F = 195.072, P = 0.000). With the prolongation of HXO-RB44 supernatant, the expression of VEGF mRNA in ARPE-19 cells gradually increased (P <0.01). The expression of VEGF protein in ARPE-19 cells was significantly increased (F = 160.687, P = 0.000) 4 h, 8 h, 12 h and 24 h after addition of HXO-RB44 cell supernatant. The expression of VEGF mRNA in ARPE- The same (P <0.05). With the prolongation of HXO-RB44 cell supernatant, the intensity of GFP in ARPE-19 cells gradually increased. Conclusion The HXO-RB44 cell supernatant can promote the expression of VEGF in ARPE-19 cells.