Modulation of early response gene expression by prostaglandins in cultured rat retinal pigment epithelium cells
Modulation of early response gene expression by prostaglandins in cultured rat retinal pigment epithelium cells
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DOI：
10.1076/ceyr.21.6.968.6987
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发表时间：
2000-01-01
影响因子：
2
通讯作者：
Bazan, NG
中科院分区：
文献类型：
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作者：
Ershov, AV;Parkins, N;Bazan, NG
目的。探讨前列腺素(PGs)对视网膜色素上皮(RPE)视杆外段(ROS)吞噬及ROS吞噬诱导基因表达的调节作用。用前列腺素E(2)、前列环素(2)、前列环素(2)α、前列环素(2)、15-脱氧-D-前列环素(12，14)-前列环素(2)或U-46619(血栓素A的稳定类似物)预先孵育原代培养的RPE细胞，并加入ROS悬液。Northern印迹法检测ZIF-268和TIS-1mRNA的表达。用凝胶迁移率改变法检测TIS-1蛋白的DNA结合活性。用酶免疫法测定组织培养上清液中PGE(2)和PGD(2)的浓度。巨噬细胞吞噬功能通过双重免疫染色结合计数和摄取的ROS计数来定量。最强的PGE(2)可显著提高基础和ROS吞噬诱导的TIS-1mRNA水平，而显著抑制基础和吞噬诱导的ZIF-268 mRNA的表达。PGD(2)、PGJ(2)和15-脱氧-Delta(12，14)-PGJ(2)可促进Ros吞噬诱导的TIS-1mRNA表达。PGF(2α)可同时诱导巨噬细胞吞噬诱导和基础TIS-1mRNA表达。U-46619和卡巴前列环素对TIS-1mRNA的表达无影响。PGE(2)是唯一影响ZIF-268表达的PG。外源前列腺素E(2)、前列环素(2)和前列环素(2)在加入浓度为1um时可显著抑制ROS的摄取，其中PGE(2)是影响ROS吞噬作用最强的PG。PGS是RPE细胞中吞噬诱导转录因子基因表达的选择性调节因子，也是ROS吞噬本身的选择性调节因子。这种调节可能有助于在RPE细胞中通过ROS吞噬受体介导的信号转导来确保靶基因差异激活的特异性。
Purpose. To explore the role of prostaglandins (PGs) as modulators of retinal pigment epithelium (RPE) rod outer segment (ROS)-phagocytosis and ROS-phagocytosis- induced gene expression.Methods. RPE cells in primary cell culture were pre-incubated with PGE(2), PGD(2), PGF(2)alpha, PGJ(2), 15-deoxy-Delta (12,14)-PGJ(2) or U-46619 (stable analog of thromboxane A(2)), and fed with a suspension of ROS. Expression of zif-268 and tis-1 mRNA was determined by Northern blotting. DNA-binding activity of TIS-1 protein was assessed by electrophoretic mobility shift assay. Concentration of PGE(2) and PGD(2) in the tissue culture medium was measured by enzyme immuno-assay. Phagocytis-tosis was quantified by counting of double-immunostained bound and ingested ROS.Results. PGE(2), the most potent of PGs, strongly elevated both basal and ROS-phagocytosis- induced levels of tis-1 mRNA, while significantly inhibiting both basal and phagocytosis-induced expression of zif-268 mRNA. PGD(2), PGJ(2) and 15-deoxy-Delta (12,14)-PGJ(2) elevated ROS-phagocytosis-induced, but not basal, expression of tis-1 mRNA expression. PGF(2 alpha) super-induced both phagocytosis-induced and basal tis-1 mRNA expression. U-46619 and carbaprostacyclin had no effect on expression of tis-1 mRNA. PGE(2) was the only PG to affect zif-268 expression. Exogenous PGE(2), PGD(2) and PGF(2), when added to the medium at 1-muM concentrations, significantly inhibited ingestion of ROS, with PGE(2) being the most potent PG affecting ROS-phagocytosis.Conclusions. PGs act as selective regulators of phagocytosis-induced transcription factor gene expression in RPE cells, as well as of ROS-phagocytosis itself. This modulation may help to ensure specificity in the differential activation of target genes by ROS-phagocytosis receptor-mediated signal transduction in RPE cells.