山羊副流感病毒3型通过Caspase途径诱导气管上皮细胞凋亡

doi:10.11843/j.issn.0366-6964.2019.10.012

Abstract

Abstract: To investigate the molecular mechanisms of apoptosis in goat bronchial epithelial cells infected with CPIV3 in vitro, the cells were inoculated with CPIV3 JS2013 strain that isolated from the previous study. The cultural supernatants of the cells were harvested at 12,24,36,48,72 and 96 h post inoculation(hpi), then qRT-PCR and TCID₅₀ assays were conducted to determine CPIV3 proliferation levels. The apoptosis was examined by Annein V-FITC/PI double staining cell apoptosis detection kit; and relevant indicators were detected by Caspase-3, Caspase-8 and Caspase-9 activity detection kit, qRT-PCR assay, and Western blot. The results showed that virus titers increased from 10^1.80 to 10^4.50 TCID₅₀·mL^-1 at 96 hpi. Observation with inverted microscope showed that CPIV3-infected cells had typical CPE, including shrink and disrupted. Flow cytometry analysis of apoptosis showed that the number of apoptotic cells increased to 19.66% at 48 hpi. Caspases activity assay demonstrated that CPIV3 infection increased the activation of Caspase-3, Caspase-8 and Caspase-9. Meanwhile, the expression levels of intrinsic pathway and extrinsic apoptotic pathway mRNA were increased. In addition, cleaved Caspase-3 was activated in CPIV3-infected cells. These findings indicate that CPIV3 infection could induce apoptosis and the activation of Caspase played important role in apoptosis.

CLC Number:

LIAO Zheng, LI Jizong, XIAO Fang, ZHONG Chunyan, YANG Leilei, MAO Li, LI Wenliang, SUN Min, ZHU Xing, JI Xinqin, ZHANG Wenwen, LIU Maojun. Caprine Parainfluenza Virus Type 3 Induces Apoptosis in Goat Bronchial Epithelial Cells via Caspase-mediated Pathways[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(10): 2061-2069.

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References 29

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Caprine Parainfluenza Virus Type 3 Induces Apoptosis in Goat Bronchial Epithelial Cells via Caspase-mediated Pathways

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