ARMS2对视网膜色素上皮细胞吞噬功能的影响

发布时间：2018-10-09 13:45

【摘要】：目的研究年龄相关性黄斑变性易感因子2（age-related maculopathy susceptibility2，ARMS2）在体外培养的人视网膜色素上皮细胞系ARPE-19及人视网膜组织中的表达情况，观察ARMS2对ARPE-19细胞吞噬功能的影响，以探讨其在年龄相关性黄斑变性发生中可能发挥的作用。方法应用RT-PCR验证ARMS2是否在ARPE-19细胞和人视网膜组织中表达。通过免疫组织化学染色法检测ARMS2在ARPE-19细胞和人视网膜组织内的表达量及表达部位。采用siRNA技术抑制ARMS2在ARPE-19细胞中的表达，，RT-PCR、免疫组化检测抑制效率；以FITC标记的乳胶微球孵育细胞，流式细胞术检测ARPE-19细胞吞噬功能；比较ARMS2被抑制前、后ARPE-19细胞在不同时间点吞噬水平的变化。结果ARMS2在ARPE-19细胞和人视网膜组织中均有表达。在ARPE-19细胞中，ARMS2表达于细胞质中。在人视网膜组织中，ARMS2表达在RPE层，其中在顶部膜的表达量最丰富。实验组（ARMS2-siRNA） ARMS2的表达水平（0.73±0.08）与正常组（1.00±0.00）及对照组（scrambled siRNA，0.95±0.13）相比明显下降，差异均有显著性，P值均小于0.05；在FITC-乳胶微球孵育后的12h、18h、24h三个时间点，实验组（ARMS2-siRNA）的荧光强度分别为38.04±1.02，68.92±0.92，78.00±0.12，对照组（scrambled siRNA）分别为77.98±5.43，94.87±0.60，98.30±0.11；实验组和对照组在12h、18h、24h三个时间点的荧光强度分别行student’s t检验，P值均小于0.01。结论ARMS2在ARPE-19细胞和人视网膜组织中均有表达。ARMS2与ARPE-19细胞的吞噬功能相关，这可能是其参与年龄相关性黄斑变性发生的机理之一。
[Abstract]:Objective to study the expression of age-related macular degeneration susceptibility factor 2 (age-related maculopathy susceptibility2,ARMS2) in cultured human retinal pigment epithelial cell line ARPE-19 and human retinal tissue, and to observe the effect of ARMS2 on the phagocytosis of ARPE-19 cells. To explore its possible role in age-related macular degeneration. Methods RT-PCR was used to verify the expression of ARMS2 in ARPE-19 cells and human retinal tissues. The expression and location of ARMS2 in ARPE-19 cells and human retina were detected by immunohistochemical staining. SiRNA technique was used to inhibit the expression of ARMS2 in ARPE-19 cells. The inhibition efficiency was detected by immunohistochemistry. The phagocytic function of ARPE-19 cells was detected by FITC labeled latex microspheres, and the phagocytosis of ARPE-19 cells was detected by flow cytometry. Changes of phagocytosis level of ARPE-19 cells at different time points. Results ARMS2 was expressed in both ARPE-19 cells and human retinal tissues. ARMS2 is expressed in the cytoplasm of ARPE-19 cells. ARMS2 is expressed in the RPE layer of human retina, especially in the parietal membrane. The expression of ARMS2 in the experimental group (0.73 卤0.08) was significantly lower than that in the normal group (1.00 卤0.08) and the control group (scrambled siRNA,0.95 卤0.13), and the difference was significant (P < 0.05). The fluorescence intensity of the experimental group (ARMS2-siRNA) was 38.04 卤1.02 卤0.92 卤0.92 卤0.92 卤78.00 卤0.12, and the (scrambled siRNA) of the control group was 77.98 卤5.43 卤0.60 卤98.30 卤0.11.The fluorescence intensity of the experimental group and the control group were measured by student's t test (P < 0.01). Conclusion the expression of ARMS2 in both ARPE-19 cells and human retinal tissues is related to the phagocytic function of ARPE-19 cells, which may be one of the mechanisms involved in age-related macular degeneration.
【学位授予单位】：济南大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R774.5

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