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首页> 外文期刊>American Journal of Physiology >In well-differentiated primary human bronchial epithelial cells, TGF-β1 and TGF-β2 induce expression of furin
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In well-differentiated primary human bronchial epithelial cells, TGF-β1 and TGF-β2 induce expression of furin

机译：在良好分化的原始人支气管上皮细胞中，TGF-β1和TGF-β2诱导Furin的表达

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摘要

The COVID-19 pandemic is an ongoing threat to public health. Since the identification of COVID-19, the disease caused by SARS-CoV-2, no drugs have been developed to specifically target SARS-CoV-2. To develop effective and safe treatment options, a better understanding of cellular mechanisms underlying SARS-CoV-2 infection is required. To fill this knowledge gap, researchers require reliable experimental systems that express the host factor proteins necessary for the cellular entry of SARS-CoV-2. These proteins include the viral receptor, angiotensin-converting enzyme 2 (ACE2), and the proteases, transmembrane serine protease 2 (TMPRSS2) and furin. A number of studies have reported cell-type-specific expression of the genes encoding these molecules. However, less is known about the protein expression of these molecules. We assessed the suitability of primary human bronchial epithelial (HBE) cells maintained in an air-liquid interface (ALI) as an experimental system for studying SARS-CoV-2 infection in vitro. During cellular differentiation, we measured the expression of ACE2, TMPRSS2, and furin over progressive ALI days by reverse transcription-quantitative polymerase chain reaction (RT-qPCR), Western blot, and immunofluorescence staining. We also explored the effect of the fibrotic cytokine TGF-(3 on the expression of these proteins in well-differentiated HBE cells. Like ACE2, TMPRSS2 and furin proteins are localized in differentiated ciliated cells, as confirmed by immunofluorescence staining. These data suggest that well-differentiated HBE cells maintained in ALI are a reliable in vitro system for investigating cellular mechanisms of SARS-CoV-2 infection. We further identified that the profibrotic mediators, TGF-(31 and TGF-02, increase the expression of furin, which is a protease required for the cellular entry of SARS-CoV-2.
机译：COVID-19流行病是对公共健康的持续威胁。由于鉴定的COVID-19，由SARS COV-2引起的疾病，没有药物已经开发专门针对SARS COV-2。为了开发有效和安全的治疗方案，需要更好地了解SARS-CoV-2感染的细胞机制。为了填补这一知识空白，研究人员需要可靠的实验系统来表达SARS-CoV-2进入细胞所必需的宿主因子蛋白。这些蛋白质包括病毒受体、血管紧张素转换酶2（ACE2）和蛋白酶、跨膜丝氨酸蛋白酶2（TMPRSS2）和furin。许多研究报告了编码这些分子的基因的细胞类型特异性表达。然而，对这些分子的蛋白质表达知之甚少。我们评估了维持在气液界面（ALI）中的原代人支气管上皮（HBE）细胞作为体外研究SARS-CoV-2感染的实验系统的适用性。在细胞分化过程中，我们通过逆转录定量聚合酶链反应（RT-qPCR）、Western blot和免疫荧光染色检测进展性ALI天数内ACE2、TMPRSS2和furin的表达。我们还探讨了纤维化细胞因子TGF的作用-（3）这些蛋白在高分化HBE细胞中的表达。免疫荧光染色证实，与ACE2一样，TMPRSS2和furin蛋白定位于分化的纤毛细胞中。这些数据表明，维持在ALI中的高分化HBE细胞是研究SARS-CoV-2感染的细胞机制的可靠体外系统e促纤维化介质TGF-（31和TGF-02）增加了furin的表达，furin是SARS-CoV-2进入细胞所需的蛋白酶。

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