The Effect of Remdesivir as An Anti-COVID-19 Drug on the Secretion of Inflammatory Markers by Chicken Liver Cells: An In Vitro Study

Zahra Akbari Jonoush, Roya Mahdavi, M. Ghafourian, S. Khoshnam, Fereshteh Nezhad Dehbashi, Maryam Farzaneh
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Abstract

Background: For severe cases of acute respiratory syndrome type 2 (SARS-CoV-2) infection (COVID-19), Remdesivir (RDV) is introduced as an anti-viral drug with side effects. Hepatotoxicity from prolonged exposure to RDV is associated with increased inflammatory factors. In this study, we evaluated the effect of RDV on the secretion of inflammatory markers by chicken liver cells. Methods: In this in vitro study, 20 stage X embryonated chicken eggs were incubated for 10 days at 37.5°C and 60–65% humidity Hamburger–Hamilton stages (stage HH35). Liver cells were grown in a medium containing DMEM/F12+10% fetal bovine serum (FBS). After three days, the culture media was supplemented with four doses of RDV (1.00, 2.00, 3.00, and 4.00 μM). After 24 and 48hs, the viability of the hepatocytes, gene expression of Sox17, CXC motif chemokine receptor 4 (CXCR4), Interleukin-1 (IL-1), Interleukin-6 (IL-6), and TNF-α, and hepatocellular functions (albumin and urea secretion) were assessed. Findings: Each hepatocyte had a prominent nucleus and a nucleolus with a hexagonal shape. The pink tint of the periodic acid Schiff (PAS) positive cells in the PAS staining verified the hepatocytes’ glycogen content. Up to 50% of the cells lose viability after 48 hours in the presence of 3 and 4 μM RDV (P<0.001). In the presence of 3 μM RDV, the production and secretion of both albumin (P<0.001) and urea (P<0.05) decreased. Besides, the expression of IL-1, IL-6, and TNF-α significantly increased after treatment with 3 μM RDV (P<0.001). Conclusion: We concluded that RDV therapy altered the expression and function of hepatocyte inflammatory factors.
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瑞德西韦抗covid -19药物对鸡肝细胞分泌炎症标志物的影响
背景:对于严重的急性呼吸综合征2型(SARS-CoV-2)感染(COVID-19)病例,瑞德西韦(Remdesivir, RDV)作为一种有副作用的抗病毒药物被引入。长期暴露于RDV的肝毒性与炎症因子增加有关。在本研究中,我们评估了RDV对鸡肝细胞炎症标志物分泌的影响。方法:采用20只X期鸡胚,在37.5℃、60-65%湿度条件下培养10 d (HH35期)。肝细胞在含有DMEM/F12+10%胎牛血清(FBS)的培养基中生长。3 d后,在培养基中添加4个剂量(1.00、2.00、3.00、4.00 μM)的RDV。24和48h后,观察肝细胞活力、Sox17、CXC基序趋化因子受体4 (CXCR4)、白细胞介素-1 (IL-1)、白细胞介素-6 (IL-6)、TNF-α基因表达及肝细胞功能(白蛋白和尿素分泌)。结果:每个肝细胞有一个突出的细胞核和一个六边形的核仁。PAS染色中周期性酸性席夫(PAS)阳性细胞呈粉红色,证实肝细胞的糖原含量。3 μM RDV和4 μM RDV作用48 h后,50%以上的细胞失去活力(P<0.001)。在3 μM RDV存在下,白蛋白(P<0.001)和尿素(P<0.05)的产生和分泌均减少。3 μM RDV处理后,IL-1、IL-6、TNF-α的表达均显著升高(P<0.001)。结论:RDV治疗可改变肝细胞炎性因子的表达和功能。
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