乙酰左旋肉碱在缺氧条件下对人视网膜色素上皮-19细胞的影响
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Fırat大学科研中心项目(No.TF:19.41)


Effect of acetyl L-carnitine on human retinal pigment epithelium-19 cells in hypoxic conditions
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Fırat University Scientific Research Project Center(No.TF:19.41)

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    摘要:

    目的:使用缺氧模型探究乙酰左旋肉碱(ALCAR)对人视网膜色素上皮(ARPE-19)细胞活力、形态完整性和血管内皮生长因子(VEGF)表达的影响。

    方法:第一组实验通过暴露ARPE-19细胞培养物于不同浓度确定最佳CoCl2剂量。建立五组ARPE-19细胞培养物,包括对照组,假手术组(200 μM CoCl2)和分别接受1、10、100 mM ALCAR联合200 μM CoCl2组,以评估ALCAR对细胞活力的影响。使用MTT法测量细胞活力。通过倒置相差显微镜观察细胞的形态特征。采用酶联免疫吸附实验(ELISA)检测ARPE-19细胞分泌VEGF和HIF-1α的水平。

    结果:ARPE-19细胞暴露于不同剂量的CoCl2中,以创建缺氧模型。然而,暴露于浓度为200 μM CoCl2时,细胞活力显著降低至83%。ALCAR在1 mM和10 mM浓度下可增加细胞活力,而最大浓度(100 mM)没有额外效果。与假手术组相比,浓度为1 mM和10 mM ALCAR组的细胞活力显著更高(P=0.041、0.019)。细胞活力和形态不受最大剂量ALCAR(100 mM)的影响。与假手术组相比,10 mM ALCAR组VEGF和HIF-1α水平显著降低(P=0.013、0.033)。

    结论:ALCAR是可行的治疗选择,可为视网膜疾病开辟新的治疗途径,与年龄相关性黄斑变性(AMD)特别相关。然而,仍需开展进一步研究明确定义其确切机制,以期充分阐明ALCAR对抗视网膜疾病的应用潜力。

    Abstract:

    AIM: To investigate the effect of acetyl-L-carnitine(ALCAR)on cell viability, morphological integrity, and vascular endothelial growth factor(VEGF)expression in human retinal pigment epithelium(ARPE-19)cells using a hypoxic model.

    METHODS: In the first set of experiments, the optimal CoCl2 dose was determined by exposing ARPE-19 cell cultures to different concentrations. To evaluate the effect of ALCAR on cell viability, five groups of ARPE-19 cell culture were established that included a control group, a sham group(200 μM CoCl2), and groups that received 1, 10 and 100 mM doses of ALCAR combined with 200 μM CoCl2, respectively. The cell viability was measured by MTT assay. The morphological characteristics of cells were observed by an inverted phase contrast microscope. The levels of VEGF and HIF-1α secretion by ARPE-19 cells were detected by enzyme linked immunosorbent assay(ELISA)assay.

    RESULTS: ARPE-19 cells were exposed to different doses of CoCl2 in order to create a hypoxia model. Nevertheless, when exposed to a concentration of 200 μM CoCl2, a notable decrease in viability to 83% was noted. ALCAR was found to increase the cell viability at 1 mM and 10 mM concentrations, while the highest concentration(100 mM)did not have an added effect. The cell viability was found to be significantly higher in the groups treated with a concentration of 1 mM and 10 mM ALCAR compared to the Sham group(P=0.041, P=0.019, respectively). The cell viability and morphology remained unaffected by the greatest dose of ALCAR(100 mM). The administration of 10 mM ALCAR demonstrated a statistically significant reduction in the levels of VEGF and HIF-1α compared with the Sham group(P=0.013, P=0.033, respectively).

    CONCLUSION: The findings from the current study indicate that ALCAR could represent a viable therapeutic option with the potential to open up novel treatment pathways for retinal diseases, particular relevance for age-related macular degeneration(AMD). However, to fully elucidate ALCAR's application potential in retinal diseases, additional investigation is necessary to clearly define the exact mechanisms involved.

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Ali Dal, Onur Çatak, Murat Erdağ,等.乙酰左旋肉碱在缺氧条件下对人视网膜色素上皮-19细胞的影响.国际眼科杂志, 2024,24(10):1515-1521.

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  • 收稿日期:2023-05-23
  • 最后修改日期:2024-06-06
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  • 在线发布日期: 2024-09-14
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