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金丝桃素抗鸡胚肾细胞传染性支气管炎病毒诱导细胞凋亡及活性氧的作用

Protective effects of hypericin against infectious bronchitis virus induced apoptosis and reactive oxygen species in chicken embryo kidney cells.

机构信息

Key Laboratory for Laboratory Animals and Comparative Medicine of Heilongjiang Province, College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China.

College of Biological and Pharmaceutical Engineering, Jilin Agriculture Science and Technology College, Jilin 132101, China.

出版信息

Poult Sci. 2019 Dec 1;98(12):6367-6377. doi: 10.3382/ps/pez465.

Abstract

Avian infectious bronchitis virus (IBV), a coronavirus, causes infectious bronchitis leading to enormous economic loss in the poultry industry worldwide. Hypericin (HY) is an excellent compound that has been investigated in antiviral, antineoplastic, and antidepressant. To investigate the inhibition effect of HY on IBV infection in chicken embryo kidney (CEK) cells, 3 different experimental designs: pre-treatment of cells prior to IBV infection, direct treatment of IBV-infected cells, and pre-treatment of IBV prior to cell infection were used. Quantitative real-time PCR (qRT-PCR), immunofluorescence assay (IFA), flow cytometry, and fluorescence microscopy were performed and virus titer was determined by TCID50. The results revealed that HY had a good anti-IBV effect when HY directly treated the IBV-infected cells, and virus infectivity decreased in a dose-dependent manner. Furthermore, HY inhibited IBV-induced apoptosis in CEK cells, and significantly reduced the mRNA expression levels of Fas, FasL, JNK, Bax, Caspase 3, and Caspase 8, and significantly increased Bcl-2 mRNA expression level in CEK cells. In addition, HY treatment could decrease IBV-induced reactive oxygen species (ROS) generation in CEK cells. These results suggested that HY showed potential antiviral activities against IBV infection involving the inhibition of apoptosis and ROS generation in CEK cells.

摘要

禽传染性支气管炎病毒(IBV),一种冠状病毒,可引起传染性支气管炎,导致全球家禽业遭受巨大的经济损失。金丝桃素(HY)是一种极好的化合物,已在抗病毒、抗肿瘤和抗抑郁方面进行了研究。为了研究 HY 对鸡胚肾(CEK)细胞中 IBV 感染的抑制作用,采用了 3 种不同的实验设计:细胞在 IBV 感染前进行预处理、IBV 感染后直接处理细胞、以及细胞感染前对 IBV 进行预处理。通过定量实时 PCR(qRT-PCR)、免疫荧光分析(IFA)、流式细胞术和荧光显微镜观察,并通过 TCID50 测定病毒滴度。结果表明,当 HY 直接处理感染 IBV 的细胞时,HY 对 IBV 具有良好的抗病毒作用,病毒感染力呈剂量依赖性下降。此外,HY 抑制了 IBV 诱导的 CEK 细胞凋亡,并显著降低了 Fas、FasL、JNK、Bax、Caspase 3 和 Caspase 8 的 mRNA 表达水平,同时显著增加了 CEK 细胞中 Bcl-2 的 mRNA 表达水平。此外,HY 处理可降低 IBV 诱导的 CEK 细胞中活性氧(ROS)的产生。这些结果表明,HY 对 IBV 感染表现出潜在的抗病毒活性,涉及抑制 CEK 细胞中的细胞凋亡和 ROS 生成。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5eac/8913760/6af5f9433a38/gr3.jpg

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