RIPK3 activation of CaMKII triggers mitochondrial apoptosis in NIBV-infected renal tubular epithelial cells

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology Pub Date : 2025-01-10 DOI:10.1016/j.vetmic.2025.110375

Ying Li , Qiurong Qi , Yifei Chen, Mengbing Ding, Manzi Huang, Cheng Huang, Ping Liu, Xiaona Gao, Xiaoquan Guo, Zhanhong Zheng

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Abstract

The purpose of this study was to investigate whether RIPK3-mediated programmed cell death can promote the replication and transmission of renal infectious bronchitis virus in renal tubular epithelial cells. Primary renal tubular epithelial cells were extracted from 1 to 7 day old Hy-Line Brown chicks, cultured in vitro by type I collagenase digestion, and infected with 1MOI SX9 strain. Cell samples were collected at 12 hpi, 24 hpi, 36 hpi and 48 hpi for experimental exploration. Our results showed that NIBV infection could lead to programmed necrosis and mitochondrial apoptosis, and the expression levels of programmed necrosis-related genes TNFR1, TRADD, FADD, RIPK1, RIPK3 and MLKL increased significantly with the extension of infection time, the expression levels of mitochondrial apoptosis-related genes Cyt-C, APAF-1, Caspase-9 and Csapase-3 were significantly increased at 36 hpi. While, after 36 hpi of virus infection, apoptosis decreased and necrosis increased, and virus replication peaked. In order to further explore the effect of necroptosis on the amplification of renal infectious virus, the RIPK3 was inhibited at 36 hpi. Inhibition of necroptosis could reduce viral replication and cell death, programmed necrosis occurred in the cells, and cell membrane perforation led to virus diffusion and replication. NIBV-induced necroptosis depends on RIPK3, and RIPK3 activates CAMKII and interacts to cause abnormal opening of mitochondrial membrane permeability transition pore, promotes Ca^2 + influx into mitochondria, initiates mitochondrial apoptosis. While, inhibition of RIPK3 significantly inhibited the programmed necrosis of cells caused by NIBV infection, making excessive necrosis into moderate necrosis, thereby inhibiting the replication of the virus in cells.

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RIPK3激活CaMKII触发nibv感染肾小管上皮细胞线粒体凋亡

本研究旨在探讨ripk3介导的程序性细胞死亡是否能促进肾传染性支气管炎病毒在肾小管上皮细胞中的复制和传播。从1 ~ 7日龄海兰褐鸡中提取原代肾小管上皮细胞，采用I型胶原酶消化法体外培养，并用1MOI SX9菌株感染。分别在12hpi、24hpi、36hpi和48hpi处采集细胞样本进行实验探索。我们的研究结果显示，NIBV感染可导致程序性坏死和线粒体凋亡，随着感染时间的延长，程序性坏死相关基因TNFR1、TRADD、FADD、RIPK1、RIPK3和MLKL的表达水平显著升高，线粒体凋亡相关基因Cyt-C、APAF-1、Caspase-9和Csapase-3的表达水平在36 hpi时显著升高。病毒感染36hpi后，细胞凋亡减少，坏死增加，病毒复制达到高峰。为了进一步探讨坏死坏死对肾感染性病毒扩增的影响，我们在36 hpi时抑制RIPK3。抑制坏死下垂可减少病毒复制和细胞死亡，细胞发生程序性坏死，细胞膜穿孔导致病毒扩散和复制。nibv诱导的坏死坏死依赖于RIPK3， RIPK3激活CAMKII并相互作用导致线粒体膜通透性过渡孔异常打开，促进Ca2 +内流线粒体，引发线粒体凋亡。而抑制RIPK3可显著抑制NIBV感染引起的细胞程序性坏死，使过度坏死变为中度坏死，从而抑制病毒在细胞中的复制。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Veterinary microbiology 农林科学-兽医学

CiteScore

5.90

自引率

6.10%

发文量

221

审稿时长

52 days

期刊介绍： Veterinary Microbiology is concerned with microbial (bacterial, fungal, viral) diseases of domesticated vertebrate animals (livestock, companion animals, fur-bearing animals, game, poultry, fish) that supply food, other useful products or companionship. In addition, Microbial diseases of wild animals living in captivity, or as members of the feral fauna will also be considered if the infections are of interest because of their interrelation with humans (zoonoses) and/or domestic animals. Studies of antimicrobial resistance are also included, provided that the results represent a substantial advance in knowledge. Authors are strongly encouraged to read - prior to submission - the Editorials (''Scope or cope'' and ''Scope or cope II'') published previously in the journal. The Editors reserve the right to suggest submission to another journal for those papers which they feel would be more appropriate for consideration by that journal. Original research papers of high quality and novelty on aspects of control, host response, molecular biology, pathogenesis, prevention, and treatment of microbial diseases of animals are published. Papers dealing primarily with immunology, epidemiology, molecular biology and antiviral or microbial agents will only be considered if they demonstrate a clear impact on a disease. Papers focusing solely on diagnostic techniques (such as another PCR protocol or ELISA) will not be published - focus should be on a microorganism and not on a particular technique. Papers only reporting microbial sequences, transcriptomics data, or proteomics data will not be considered unless the results represent a substantial advance in knowledge. Drug trial papers will be considered if they have general application or significance. Papers on the identification of microorganisms will also be considered, but detailed taxonomic studies do not fall within the scope of the journal. Case reports will not be published, unless they have general application or contain novel aspects. Papers of geographically limited interest, which repeat what had been established elsewhere will not be considered. The readership of the journal is global.