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Acute mechanical stress in primary porcine RPE cells induces angiogenic factor expression and in vitro angiogenesis.
Journal of Biological Engineering ( IF 5.6 ) Pub Date : 2020-04-25 , DOI: 10.1186/s13036-020-00235-4
Farhad Farjood 1, 2 , Amir Ahmadpour 1, 3 , Sassan Ostvar 4 , Elizabeth Vargis 1
Affiliation  

Background Choroidal neovascularization (CNV) is a major cause of blindness in patients with age-related macular degeneration. CNV is characterized by new blood vessel growth and subretinal fluid accumulation, which results in mechanical pressure on retinal pigment epithelial (RPE) cells. The overexpression of RPE-derived angiogenic factors plays an important role in inducing CNV. In this work, we investigated the effect of mechanical stress on the expression of angiogenic factors in porcine RPE cells and determined the impact of conditioned medium on in-vitro angiogenesis. Results The goal of this study was to determine whether low levels of acute mechanical stress during early CNV can induce the expression of angiogenic factors in RPE cells and accelerate angiogenesis. Using a novel device, acute mechanical stress was applied to primary porcine RPE cells and the resulting changes in the expression of major angiogenic factors, VEGF, ANG2, HIF-1α, IL6, IL8 and TNF-α, were examined using immunocytochemistry, qRT-PCR, and ELISA. An in vitro tube formation assay was used to determine the effect of secreted angiogenic proteins due to mechanical stress on endothelial tube formation by human umbilical vein endothelial cells (HUVECs). Our results showed an increase in the expression of VEGF, ANG2, IL-6 and IL-8 in response to mechanical stress, resulting in increased in vitro angiogenesis. Abnormal epithelial-mesenchymal transition (EMT) in RPE cells is also associated with CNV and further retinal degeneration. Our qRT-PCR results verified an increase in the expression of EMT genes, CDH2, VIM and FN1, in RPE cells. Conclusions In conclusion, we showed that acute mechanical stress induces the expression of major angiogenic and EMT factors and promotes in vitro angiogenesis, suggesting that mechanical stress plays a role in promoting aberrant angiogenesis in AMD.

中文翻译:

原代猪 RPE 细胞中的急性机械应力诱导血管生成因子表达和体外血管生成。

背景脉络膜新生血管(CNV)是老年性黄斑变性患者失明的主要原因。CNV 的特点是新血管生长和视网膜下液积聚,这导致视网膜色素上皮 (RPE) 细胞受到机械压力。RPE衍生的血管生成因子的过表达在诱导CNV中起重要作用。在这项工作中,我们研究了机械应力对猪 RPE 细胞中血管生成因子表达的影响,并确定了条件培养基对体外血管生成的影响。结果本研究的目的是确定早期CNV期间低水平的急性机械应力是否可以诱导RPE细胞中血管生成因子的表达并加速血管生成。使用新颖的设备,对原代猪 RPE 细胞施加急性机械应力,并使用免疫细胞化学、qRT-PCR 和 ELISA 检测主要血管生成因子 VEGF、ANG2、HIF-1α、IL6、IL8 和 TNF-α 表达的变化。体外管形成测定用于确定由于机械应力对人脐静脉内皮细胞 (HUVEC) 内皮管形成的分泌血管生成蛋白的影响。我们的结果显示,VEGF、ANG2、IL-6 和 IL-8 的表达在机械应力下增加,导致体外血管生成增加。RPE 细胞中的异常上皮间质转化 (EMT) 也与 CNV 和进一步的视网膜变性有关。我们的 qRT-PCR 结果证实了 RPE 细胞中 EMT 基因 CDH2、VIM 和 FN1 的表达增加。结论 总之,
更新日期:2020-04-25
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