【摘要】：目的研究年齡相關(guān)性黃斑變性易感因子2（age-related maculopathy susceptibility2，ARMS2）在體外培養(yǎng)的人視網(wǎng)膜色素上皮細(xì)胞系A(chǔ)RPE-19及人視網(wǎng)膜組織中的表達(dá)情況，觀察ARMS2對ARPE-19細(xì)胞吞噬功能的影響，以探討其在年齡相關(guān)性黃斑變性發(fā)生中可能發(fā)揮的作用。方法應(yīng)用RT-PCR驗(yàn)證ARMS2是否在ARPE-19細(xì)胞和人視網(wǎng)膜組織中表達(dá)。通過免疫組織化學(xué)染色法檢測ARMS2在ARPE-19細(xì)胞和人視網(wǎng)膜組織內(nèi)的表達(dá)量及表達(dá)部位。采用siRNA技術(shù)抑制ARMS2在ARPE-19細(xì)胞中的表達(dá)，，RT-PCR、免疫組化檢測抑制效率；以FITC標(biāo)記的乳膠微球孵育細(xì)胞，流式細(xì)胞術(shù)檢測ARPE-19細(xì)胞吞噬功能；比較ARMS2被抑制前、后ARPE-19細(xì)胞在不同時(shí)間點(diǎn)吞噬水平的變化。結(jié)果ARMS2在ARPE-19細(xì)胞和人視網(wǎng)膜組織中均有表達(dá)。在ARPE-19細(xì)胞中，ARMS2表達(dá)于細(xì)胞質(zhì)中。在人視網(wǎng)膜組織中，ARMS2表達(dá)在RPE層，其中在頂部膜的表達(dá)量最豐富。實(shí)驗(yàn)組（ARMS2-siRNA） ARMS2的表達(dá)水平（0.73±0.08）與正常組（1.00±0.00）及對照組（scrambled siRNA，0.95±0.13）相比明顯下降，差異均有顯著性，P值均小于0.05；在FITC-乳膠微球孵育后的12h、18h、24h三個(gè)時(shí)間點(diǎn)，實(shí)驗(yàn)組（ARMS2-siRNA）的熒光強(qiáng)度分別為38.04±1.02，68.92±0.92，78.00±0.12，對照組（scrambled siRNA）分別為77.98±5.43，94.87±0.60，98.30±0.11；實(shí)驗(yàn)組和對照組在12h、18h、24h三個(gè)時(shí)間點(diǎn)的熒光強(qiáng)度分別行student’s t檢驗(yàn)，P值均小于0.01。結(jié)論ARMS2在ARPE-19細(xì)胞和人視網(wǎng)膜組織中均有表達(dá)。ARMS2與ARPE-19細(xì)胞的吞噬功能相關(guān)，這可能是其參與年齡相關(guān)性黃斑變性發(fā)生的機(jī)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 鹵0.08) was significantly lower than that in the normal group (1.00 鹵0.08) and the control group (scrambled siRNA,0.95 鹵0.13), and the difference was significant (P < 0.05). The fluorescence intensity of the experimental group (ARMS2-siRNA) was 38.04 鹵1.02 鹵0.92 鹵0.92 鹵0.92 鹵78.00 鹵0.12, and the (scrambled siRNA) of the control group was 77.98 鹵5.43 鹵0.60 鹵98.30 鹵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.
【學(xué)位授予單位】：濟(jì)南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R774.5

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