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[目的]研究糖基化终末产物对视网膜色素上皮细胞和T淋巴细胞中的免疫调控因子的作用,深入理解糖基化终末产物对糖尿病视网膜病变的免疫机制。[方法]利用荧光定量PCR技术,测定在不同浓度的AGEs(0,10,50,100,500μg/ml)培养基中体外培养的视网膜色素上皮细胞和T淋巴细胞产生的免疫调控因子的mRNA水平。利用Western-Blot技术,测定在不同浓度的AGEs(0,10,50,100,500μg/ml)培养基中体外培养的视网膜小胶质细胞和T淋巴细胞产生的免疫调控因子的蛋白水平。[结果]经测定,视网膜色素上皮细胞中CD80,MHCII,I-CAM-1,CD86的mRNA表达量上升,差异有统计学意义(P﹤0.05),与AGEs的浓度呈正相关。淋巴细胞中的CD69,IL-8,MCP-1的表达量上升,差异有统计学意义(P﹤0.05),与AGEs的浓度呈正相关。在100μg/ml浓度的AGEs培养条件下,与色素上皮细胞共培养的T淋巴细胞中的CD69,IL-8,MCP-1表达量,相对T细胞在100μg/mlAGEs浓度下将单独培养的水平的上升更为显著。[结论]在AGEs模拟的糖尿病环境中,色素上皮细胞的免疫活性显著提高,继而激活T淋巴细胞,成为糖尿病视网膜病病原发病机制之一。
[Objective] To investigate the effect of advanced glycation end products on immune regulators in retinal pigment epithelial cells and T lymphocytes, and to understand the immune mechanism of advanced glycation end products on diabetic retinopathy. [Method] Fluorescent quantitative PCR was used to determine the mRNA level of immunoregulatory cytokines produced by cultured retinal pigment epithelial cells and T lymphocytes in different concentrations of AGEs (0, 10, 50, 100 and 500 μg / ml). Protein levels of immunomodulatory factors produced by retinal microglial cells and T lymphocytes cultured in vitro in different concentrations of AGEs (0, 10, 50, 100, 500 μg / ml) were determined by Western-Blot technique. [Result] The mRNA expression of CD80, MHCII, I-CAM-1 and CD86 in retinal pigment epithelial cells increased, the difference was statistically significant (P <0.05), and the concentration of AGEs was positively correlated. The expression of CD69, IL-8 and MCP-1 in lymphocytes increased with a statistically significant difference (P <0.05), which was positively correlated with the concentration of AGEs. The levels of CD69, IL-8, and MCP-1 expression in T lymphocytes co-cultured with the pigment epithelial cells at a concentration of 100 μg / ml of AGEs cultured at a concentration of 100 μg / ml AGS relative to the T cells at a culture medium alone The rise is even more pronounced. [Conclusion] In the diabetic environment with AGEs, the immunopotentiation of pigment epithelial cells was significantly increased, which in turn activated T lymphocytes and became one of the pathogenesis of diabetic retinopathy.