人羊水处理后人视网膜色素上皮细胞中及其受体的表达增强

Augmented Expression of and Its Receptors in Human Retinal Pigment Epithelial Cells Following Treatment with Human Amniotic Fluid.

作者信息

Safdari Bahar, Rezaei-Kanavi Mozhgan, Mishan Mohammad Amir, Ahmadieh Hamid, Soheili Zahra-Soheila, Salahshourifar Iman, Suri Fatemeh

机构信息

Ocular Tissue Engineering Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Ophthalmic Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

出版信息

Iran J Public Health. 2022 Jul;51(7):1658-1666. doi: 10.18502/ijph.v51i7.10100.

DOI:10.18502/ijph.v51i7.10100

PMID:36248281

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9529728/

Abstract

BACKGROUND

Nogo-A, a myelin-associated inhibitor for neurite outgrowth, has important role in visual system development. Trans-differentiation ability of human amniotic fluid (HAF) on human retinal pigment epithelial cells (hRPEs) towards neural progenitor cells has been observed in several studies. We aimed to investigate the expression of gene and its receptors as a marker of neural differentiation in HAF-treated hRPE cells.

METHODS

hRPE cells were cultivated and immune characterized via RPE65 and cytokeratin 8/18 protein markers. Also, the cytotoxicity effect of 30% HAF on hRPE cells was evaluated using ELISA cell death assay. Finally, expression of and its receptors, and was evaluated in the cells treated with HAF in comparison with FBS-treated cells using quantitative real-time PCR.

RESULTS

Harvested cells showed immunoreactivity for cytokeratin 8/18 and RPE65, confirming the hRPE cell identity. Besides, HAF had no cytotoxic effect on hRPE cells compared with FBS-treated cells. Results showed that and its receptors were expressed in cultured hRPE cells. Besides, comparative gene expression analysis revealed significant increased expression of the investigated genes in HAF-treated hRPE cells compared to FBS-treated cells.

CONCLUSION

Augmented expression of and its receptors can support neural differentiation of hRPE when the cells are treated with HAF. Our outcomes provide more evidences on the trans-differentiation ability of HAF on hRPE cells into neural progenitors and retinal neural cells, but further studies are needed to elucidate the exact mechanism.

摘要

背景

Nogo-A是一种神经突生长的髓磷脂相关抑制剂，在视觉系统发育中起重要作用。多项研究观察到人类羊水（HAF）对人视网膜色素上皮细胞（hRPE）向神经祖细胞的转分化能力。我们旨在研究该基因及其受体的表达，作为HAF处理的hRPE细胞中神经分化的标志物。

方法

培养hRPE细胞，并通过RPE65和细胞角蛋白8/18蛋白标志物进行免疫鉴定。此外，使用ELISA细胞死亡测定法评估30% HAF对hRPE细胞的细胞毒性作用。最后，与FBS处理的细胞相比，使用定量实时PCR评估HAF处理的细胞中该基因及其受体的表达。

结果

收获的细胞对细胞角蛋白8/18和RPE65显示免疫反应性，证实了hRPE细胞的身份。此外，与FBS处理的细胞相比，HAF对hRPE细胞没有细胞毒性作用。结果表明，该基因及其受体在培养的hRPE细胞中表达。此外，比较基因表达分析显示，与FBS处理的细胞相比，HAF处理的hRPE细胞中所研究基因的表达显著增加。