Spraul C W, Kaven C K, Kampmeier J K, Lang G K, Lang G E
University of Ulm, Department of Ophthalmology Ulm, Germany.
Curr Eye Res. 1999 Dec;19(6):483-90. doi: 10.1076/ceyr.19.6.483.5281.
The retinal pigment epithelium (RPE) is involved in the pathogenesis of age-related macular degeneration. The purpose of this study was to assess the effect of thalidomide, octreotide, and prednisolone on the proliferation and migration of bovine RPE cells in vitro.
The migration assay was performed in double-chamber-wells separated by a membrane filter with 8 microm pores. Cells were allowed to migrate vertically for 7 hr, afterwards the cells on both filtersides were fixed, stained, and the migrated cells were counted. To examine RPE proliferation, bovine RPE cells were seeded subconfluently followed by an incubation with octreotide, thalidomide or prednisolone in a concentration gradient for 24 hr. Stimulation or inhibition of DNA synthesis was measured by [(3)H]-thymidine incorporation. Statistical analysis was performed with the paired student's t-test.
Statistically significant (p < 0.05) inhibition of RPE cell proliferation was measured for thalidomide at a concentration of 10-50 microg/ml, for octreotide at a concentration of 5 x 10(-4) and 5 x 10(-5) M, and for prednisolone at a concentration of 250 and 500 microg/ml as compared to the negative control. RPE cell migration was significantly (p < 0.05) inhibited by thalidomide at a concentration of 10 microg/ml, by octreotide at a concentration of 5 x 10(-5) M, and also by prednisolone at a concentration of 500 microg/ml as compared to the negative control.
Although the main effect of thalidomide, octreotide, and prednisolone when treating patients with choroidal neovascular membranes is probably related to the inhibition of angiogenesis it should be kept in mind that these substances may additionally inhibit RPE proliferation and migration.
视网膜色素上皮(RPE)参与年龄相关性黄斑变性的发病机制。本研究旨在评估沙利度胺、奥曲肽和泼尼松龙对体外培养的牛RPE细胞增殖和迁移的影响。
迁移实验在由8微米孔径的膜滤器分隔的双室孔板中进行。细胞垂直迁移7小时,之后对滤器两侧的细胞进行固定、染色,并对迁移的细胞进行计数。为检测RPE细胞增殖,将牛RPE细胞以亚汇合状态接种,随后用不同浓度梯度的奥曲肽、沙利度胺或泼尼松龙孵育24小时。通过[³H] - 胸腺嘧啶核苷掺入法测定DNA合成的刺激或抑制情况。采用配对学生t检验进行统计学分析。
与阴性对照相比,沙利度胺浓度为10 - 50微克/毫升、奥曲肽浓度为5×10⁻⁴和5×10⁻⁵摩尔/升、泼尼松龙浓度为250和500微克/毫升时,RPE细胞增殖受到统计学显著(p < 0.05)抑制。与阴性对照相比,沙利度胺浓度为10微克/毫升、奥曲肽浓度为5×10⁻⁵摩尔/升、泼尼松龙浓度为500微克/毫升时,RPE细胞迁移受到显著(p < 0.05)抑制。
尽管沙利度胺、奥曲肽和泼尼松龙在治疗脉络膜新生血管膜患者时的主要作用可能与抑制血管生成有关,但应记住这些物质可能还会抑制RPE细胞的增殖和迁移。