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首页> 外文期刊>Investigative ophthalmology & visual science >Alkylphosphocholines inhibit proliferation of human retinal pigment epithelial cells.
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Alkylphosphocholines inhibit proliferation of human retinal pigment epithelial cells.

机译：烷基磷酸胆碱抑制人视网膜色素上皮细胞的增殖。

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摘要

PURPOSE: To investigate the effect and mechanism of action of alkylphosphocholines (APCs) on proliferation of human retinal pigment epithelium (RPE) cells and RPE-mediated collagen matrix contraction in vitro. METHODS: Cultured RPE cells of five human donors were treated with four APCs in the presence of fetal calf serum. Proliferation was assessed by the tetrazolium dye-reduction (MTT) assay and by counting the number of cells dividing in culture. The effect of APCs on RPE-mediated matrix contraction was determined in three-dimensional collagen gels. Cell viability was tested by the trypan blue exclusion assay. As a possible mechanism of APC action, protein kinase C (PKC) activity was quantified by scintillation counting of (32)P-labeled phosphate transferred to a PKC-specific substrate. RESULTS: All APCs inhibited RPE proliferation and RPE-mediated collagen matrix contraction in a dose-dependent manner in vitro. The antiproliferative and anticontractile effect of APCs increased with elongation of thefatty acid chain beyond C20. IC(50)s of all APCs varied between 8.5 micro M (erucyl-phosphocholine, C22:1-PC), 9.0 micro M (Z)-12-heneicosenyl-phosphocholine, C21:1-PC), 11.0 micro M (Z)-10-eicosenyl-phosphocholine, C20:1-PC), and 26.5 micro M (oleyl-phosphocholine, C18:1-PC). Trypan blue staining revealed a toxicity below 5% for all APCs within the concentration interval tested. PKC activity was significantly reduced by all four APCs, with C22:1-PC being the most effective. CONCLUSIONS: APCs inhibit proliferation of RPE cells and RPE-mediated matrix contraction in vitro at nontoxic concentrations. This effect seems to be exerted through inhibition of PKC activity. Therefore, APCs are promising candidates for treatment of RPE-mediated proliferative processes such as proliferative vitreoretinopathy.
机译：目的：探讨烷基磷酸胆碱（APCs）在体外对人视网膜色素上皮细胞（RPE）增殖和RPE介导的胶原基质收缩的作用及其作用机理。方法：在胎牛血清存在的情况下，用四种APC处理五名人类供体的RPE细胞。通过四唑鎓染料还原（MTT）测定并通过计数在培养物中分裂的细胞数来评估增殖。在三维胶原凝胶中确定了APC对RPE介导的基质收缩的影响。通过台盼蓝排除试验测试细胞活力。作为APC作用的可能机制，通过闪烁计数转移至PKC特异性底物的（32）P标记的磷酸盐来定量蛋白激酶C（PKC）活性。结果：所有APCs在体外均以剂量依赖的方式抑制RPE增殖和RPE介导的胶原基质收缩。 APCs的抗增殖和抗收缩作用随C20上方脂肪酸链的延长而增加。所有APC的IC（50）在8.5 micro M（芥子酰基磷酸胆碱，C22：1-PC），9.0 micro M（Z）-12-二十碳烯基磷酸胆碱，C21：1-PC）和11.0 micro M（Z ）-10-二十碳烯基磷酸胆碱（C20：1-PC）和26.5 micro M（油基磷酸胆碱，C18：1-PC）。台盼蓝染色显示，在所测试的浓度范围内，所有APC的毒性均低于5％。四种APC均显着降低PKC活性，其中C22：1-PC最有效。结论：APCs在无毒浓度下可抑制RPE细胞的增殖和RPE介导的基质收缩。这种作用似乎是通过抑制PKC活性发挥的。因此，APCs是治疗RPE介导的增生过程（如增生性玻璃体视网膜病变）的有希望的候选者。

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