牛病毒性腹泻病毒诱导铁死亡对其复制水平的影响

doi:10.11843/j.issn.0366-6964.2024.12.034

Abstract

Abstract:

This study aimed to investigate the regulatory effect of bovine viral diarrhoea virus(BVDV) infection on ferroptosis in Madin-Darby Bovine Kidney(MDBK) cells and its effect on viral replication. Technologies such as confocal microscopy, Western blot, qPCR and electron microscopy were used to detect ferroptosis and the level of viral replication in BVDV-infected cells. Furthermore, the viral replication level and inflammatory cytokines expression in BVDV-infected cells pretreated with Fer-1, a commonly used ferroptosis inhibitor, were also detected. The results showed that, compared with the normal control group, the mortality rate of cells infected with BVDV (MOI=5) was significantly increased (P < 0.05), which was accompanied with increased levels of Fe²⁺ and lipid peroxidation (P < 0.05). Transmission electron microscopy (TEM) analysis revealed that BVDV-infected and Erastin-treated cells displayed shrunk mitochondria with fewer cristae, increased mitochondrial membrane density and decreased mitochondrial mean areas compared with those of mock-infected cells, which was a typical morphological feature of ferroptosis. Fer-1 treatment significantly inhibited the levels of viral replication in BVDV-infected cells (P < 0.05). In addition, ferroptosis induced by BVDV infection had positive regulatory effects on inflammatory cytokines expression, including IL-1β, IL-18 and IFN-β. The results suggested that BVDV infection can induce ferroptosis in host cells, and the induction of ferroptosis enhanced the level of viral replication and the expression of inflammatory cytokines.

Key words: bovine viral diarrhoea virus (BVDV), ferroptosis, virus replication, inflammatory cytokines

CLC Number:

S852.653

ZHANG Zixuan, ZHANG Ying, LI Zhijun, YANG Jingling, JIANG Zihao, HUANG Huamin, QI Xuefeng. The Effects of Bovine Viral Diarrhoea Virus (BVDV)-induced Ferroptosis on Virus Replication[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5716-5724.

Figures/Tables 10

Table 1

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Fig. 6

Fig. 7

Fig. 8

Fig. 9

References 23

1	韩猛立, 张倩, 赵文娟, 等. 牛病毒性腹泻病毒的分子生物学特性及发病机制研究进展[J]. 中国畜牧兽医, 2023, 50 (11): 4632- 4645.
	HAN M L , ZHANG Q , ZHAO W J , et al. Advances on molecular biology and pathogenesis of bovine viral diarrhea virus[J]. China Animal Husbandry & Veterinary Medicine, 2023, 50 (11): 4632- 4645.
2	李新培, 周伟光, 关平原, 等. 牛病毒性腹泻病毒致病机制研究进展[J]. 中国畜牧兽医, 2018, 45 (8): 2303- 2311.
	LI X P , ZHOU W G , GUAN P Y , et al. Research progress on pathogenesis of bovine viral diarrhoea virus[J]. China Animal Husbandry & Veterinary Medicine, 2018, 45 (8): 2303- 2311.
3	张天生. 牛病毒性腹泻病毒的病原学、致病机制及免疫反应的研究进展[J]. 中国动物保健, 2023, 25 (11): 40- 41. doi: 10.3969/j.issn.1008-4754.2023.11.020
	ZHANG T S . Research progress on the etiology, pathogenic mechanism and immunoreaction of bovine viral diarrhea virus[J]. China Animal Health, 2023, 25 (11): 40- 41. doi: 10.3969/j.issn.1008-4754.2023.11.020
4	管文斌, 刘新光, 赵炜. 铁死亡与衰老[J]. 中国生物化学与分子生物学报, 2024, 40 (1): 28- 37.
	GUAN W B , LIU X G , ZHAO W . Ferroptosis and aging[J]. Chinese Journal of Biochemistry and Molecular Biology, 2024, 40 (1): 28- 37.
5	CHEN J , FU J P , ZHAO S , et al. Free radical and viral infection: a review from the perspective of ferroptosis[J]. Vet Sci, 2023, 10 (7): 456. doi: 10.3390/vetsci10070456
6	CHENG J H , TAO J , LI B Q , et al. Swine influenza virus triggers ferroptosis in A549 cells to enhance virus replication[J]. Virol J, 2022, 19 (1): 104. doi: 10.1186/s12985-022-01825-y
7	BANERJEE S , SARKAR R , MUKHERJEE A , et al. Rotavirus-induced lncRNA SLC7A11-AS1 promotes ferroptosis by targeting cystine/glutamate antiporter xCT (SLC7A11) to facilitate virus infection[J]. Virus Res, 2024, 339, 199261. doi: 10.1016/j.virusres.2023.199261
8	LI Y G , BAO Y W , LI Y , et al. RSL3 inhibits porcine epidemic diarrhea virus replication by activating ferroptosis[J]. Viruses, 2023, 15 (10): 2080. doi: 10.3390/v15102080
9	CHEN Y , GUO Y C , CHANG H , et al. Brequinar inhibits African swine fever virus replication in vitro by activating ferroptosis[J]. Virol J, 2023, 20 (1): 242. doi: 10.1186/s12985-023-02204-x
10	FU Q , SHI H J , ZHANG H , et al. Autophagy during early stages contributes to bovine viral diarrhea virus replication in MDBK cells[J]. J Basic Microbiol, 2014, 54 (10): 1044- 1052. doi: 10.1002/jobm.201300750
11	COUSSENS P M , VERMAN N , COUSSENS M A , et al. Cytokine gene expression in peripheral blood mononuclear cells and tissues of cattle infected with Mycobacterium avium subspparatuberculosis: evidence for an inherent proinflammatory gene expression pattern[J]. Infect Immun, 2004, 72 (3): 1409- 1422. doi: 10.1128/IAI.72.3.1409-1422.2004
12	谌婕, 左之才, 才冬杰, 等. 牛病毒性腹泻病毒全球基因型与亚型流行情况[J]. 浙江农业学报, 2022, 34 (12): 2622- 2628. doi: 10.3969/j.issn.1004-1524.2022.12.05
	CHEN J , ZUO Z C , CAI D J , et al. Global genotype and subgenotype prevalence of bovine viral diarrhea virus[J]. Acta Agriculturae Zhejiangensis, 2022, 34 (12): 2622- 2628. doi: 10.3969/j.issn.1004-1524.2022.12.05
13	MAURER K , KREY T , MOENNIG V , et al. CD46 is a cellular receptor for bovine viral diarrhea virus[J]. J Virol, 2004, 78 (4): 1792- 1799. doi: 10.1128/JVI.78.4.1792-1799.2004
14	侯佩莉, 王洪梅, 赵贵民, 等. 牛病毒性腹泻病毒致病分子机制的研究进展[J]. 内蒙古大学学报(自然科学版), 2017, 48 (4): 452- 457.
	HOU P L , WANG H M , ZHAO G M , et al. Research advances in the molecular mechanism of bovine viral diarrhea virus[J]. Journal of Inner Mongolia University (Natural Science Edition), 2017, 48 (4): 452- 457.
15	周文博, 孔晨飞, 秦高伟, 等. 铁死亡发生机制的研究进展[J]. 生物化学与生物物理进展, 2018, 45 (1): 16- 22.
	ZHOU W B , KONG C F , QIN G W , et al. Research progresss on mechanism of ferroptosis[J]. Progress in Biochemistry and Biophysics, 2018, 45 (1): 16- 22.
16	刘利, 杜旅, 张平, 等. 线粒体参与铁死亡的研究进展[J]. 现代医药卫生, 2023, 39 (20): 3541- 3546. doi: 10.3969/j.issn.1009-5519.2023.20.026
	LIU L , DU L , ZHANG P , et al. Research progress of mitochondria involved in ferroptosis[J]. Journal of Modern Medicine & Health, 2023, 39 (20): 3541- 3546. doi: 10.3969/j.issn.1009-5519.2023.20.026
17	FOO J , BELLOT G , PERVAIZ S , et al. Mitochondria-mediated oxidative stress during viral infection[J]. Trends Microbiol, 2022, 30 (7): 679- 692. doi: 10.1016/j.tim.2021.12.011
18	KAJARABILLE N , LATUNDE-DADA G O . Programmed cell-death by ferroptosis: antioxidants as mitigators[J]. Int J Mol Sci, 2019, 20 (19): 4968. doi: 10.3390/ijms20194968
19	李易聪, 蒲飞洋, 冯茜莉, 等. 牛病毒性腹泻病毒蛋白的免疫学特性以及相关疫苗研究进展[J]. 畜牧兽医学报, 2023, 54 (4): 1381- 1391. doi: 10.11843/j.issn.0366-6964.2023.04.004
	LI Y C , PU F Y , FENG X L , et al. Research progress on immunological characteristics of bovine viral diarrhea virus protein and vaccines[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54 (4): 1381- 1391. doi: 10.11843/j.issn.0366-6964.2023.04.004
20	张桂洪, 唐旭东. 铁死亡与免疫细胞关系的研究进展[J]. 中国比较医学杂志, 2023, 33 (3): 90- 97.
	ZHANG G H , TANG X D . Research progress on the relationship between ferroptosis and immune cells[J]. Chinese Journal of Comparative Medicine, 2023, 33 (3): 90- 97.
21	王浩, 肖金龙, 沈珏, 等. 细胞死亡的新方式——铁死亡与铜死亡[J]. 畜牧兽医学报, 2024, 55 (2): 461- 470. doi: 10.11843/j.issn.0366-6964.2024.02.005
	WANG H , XIAO J L , SHEN J , et al. New ways of cell death—ferroptosis and cuproptosis[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55 (2): 461- 470. doi: 10.11843/j.issn.0366-6964.2024.02.005
22	黄家望, 马心悦, 冯芷莹, 等. A型流感病毒通过激活HIF-1α/iNOS/VEGF信号通路诱导肺上皮细胞铁死亡的机制[J]. 微生物学通报, 2024, 51 (1): 306- 322.
	HUANG J W , MA X Y , FENG Z Y , et al. Mechanism of influenza A virus inducing ferroptosis in lung epithelial cells by activating the HIF-1α/iNOS/VEGF signaling pathway[J]. Microbiology China, 2024, 51 (1): 306- 322.
23	LIU C C , WU X H , BING X , et al. H1N1 influenza virus infection through NRF2-KEAP1-GCLC pathway induces ferroptosis in nasal mucosal epithelial cells[J]. Free Radic Biol Med, 2023, 204, 226- 242. doi: 10.1016/j.freeradbiomed.2023.05.004

基因名称 Genes name	引物 Primer	序列 Sequence	登录号 Accession No.	长度/bp Length
BVDV N^pro	F	CAACGCTAAAACTGCCACATAA	JX065783	124
BVDV N^pro	R	CCCTGGTTTTAAATAGATTCCACTC	JX065783	124
IFN-β^[10]	F	CCTGTGCCTGATTTCATCATGA	NM_174350	97
IFN-β^[10]	R	GCAAGCTGTAGCTCCTGGAAAG	NM_174350	97
IL-18^[11]	F	GAAAATGATGAAGACCTGGAATCA	NM_174091	84
IL-18^[11]	R	AACTTGGTCATTCAATTTCGTATGA	NM_174091	84
IL-1β^[11]	F	GCCTTCAATAACTGTGGAACCAAT	NM_174093	80
IL-1β^[11]	R	GTATATTTCAGGCTTGGTGAAAGGA	NM_174093	80
GAPDH^[10]	F	GGCAAGTTCAACGGCACAG	NM_001034034	248
GAPDH^[10]	R	TTCACGCCCATCACAAACA	NM_001034034	248

[1]	Hongyu FU, Yue LI, Han CUI, Jiuzhi LI, Wanxue XU, Xi WANG, Ruifeng FAN. The Mechanism of Long-Chain acyl-CoA Synthetase 4-mediated Ferroptosis [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(9): 3792-3801.
[2]	Yiqian FU, Dongge LIANG, Mingyang WANG, Jiajia PAN, Yanbin YANG, Lei ZENG, Xiangtao KANG. Construction of Interferon Regulatory Factor Knockdown Cell Line and Its Effect on Pseudorabies Virus Proliferation [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(9): 4100-4109.
[3]	ZHANG Xinting, QIU Wenyue, PANG Xiaoyue, SU Yiman, YE Jiali, HUANG Jianjia, ZHOU Shuilian, TANG Zhaoxin, WANG Rongmei, SU Rongsheng. Effect of Asiatic Acid Alleviating Myocardial Injury Caused by Lipopolysaccharide through Inhibiting Oxidative Stress and Ferroptosis in Broilers [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1787-1799.
[4]	WANG Hao, XIAO Jinlong, SHEN Jue, ZHAO Jingang, WANG Shuai, LIU Gen, ZHAO Ru, XIAO Peng, GAO Hong. New Ways of Cell Death—Ferroptosis and Cuproptosis [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 461-470.
[5]	LIU Weijiao, HE Qing, JIANG Yifan, CAO Siyu, ZHANG Shirui, LÜ Jialu, LONG Ping, YANG Lingchen, ZHOU Chuan, WANG Naidong. The Role of Porcine Lck Protein in PCV2 Replication in vitro [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5751-5761.
[6]	XIONG Zhiwei, WANG Yun, CAO Huabin, PENG Chengcheng, YANG Fan, DAI Xueyan, XING Chenghong, LIU Lingli, LI Jingni, HU Aiming. Molybdenum and Cadmium Combined Exposure Mediates Oxidative Stress and Ferroptosis Induced Kidney Damage in Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5802-5812.
[7]	Kexin WANG, Xian WU, Haizhou GONG, Qiaoyu FANG, Xiangchen LI, Yanan ZHANG. Effects of Co-Treatment of Sodium Butyrate and Indole-3-Propionic Acid on Tight Junctions and Inflammatory Cytokines in Caco-2 Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(11): 5124-5134.
[8]	SONG Wenyan, ZHANG Hanwen, WU Aodi, ZHANG Liyan, LIU Zhao, YE Tongtong, CHEN Chuangfu, SHENG Jinliang. Screening of Nanobodies against Porcine Reproductive and Respiratory Syndrome Virus GP5 Protein and Exploration of Their Inhibitory Effect on Virus Replication [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 258-270.
[9]	MAO Peng, WANG Zhihao, LI Jianji, CUI Luying, ZHU Guoqiang, MENG Xia, DONG Junsheng, WANG Heng. Research Progress of Ferroptosis in Bacterial Infection [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2280-2287.
[10]	FANG Yuan, HOU Qiaodi, XIANG Chaohui, ZHAO Hongyi, QI Xuefeng. Regulatory Effects of IFITM3 on Proliferation of Peste des Petits Ruminants Virus (PPRV) in Goat Endometrial Epithelial Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2200-2207.
[11]	LIU Yankun, LUO Runbo, LIN Yan, ZHU Weiyun. Effects of Phage Cocktail on Growth Performance, Blood Parameters and Fecal Microbiota of Weaned Piglets [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1555-1567.
[12]	CHEN Jingyi, YU Miao, ZHANG Jinyang, FAN Kun, YANG Guijun, GE Ming, ZHANG Ruili. Study on the Involvement of Ferroptosis in Liver Injury of Cadmium-exposed Chickens [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(2): 787-802.
[13]	CHEN Danian, MA Xusheng, DAI Junfei, WANG Yang, LI Qian, BAI Heng, MAO Tiantian, LIU Yongsheng, DING Long, CHEN Haohan, CHEN Siyan, RAO Yufei, JIA Ning, ZHANG Jie, ZHENG Haixue, LIU Xiangtao. Preliminary Study on the Function of MGF360-13L Gene of African Swine Fever Virus Multigene Family [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(12): 4419-4428.
[14]	DUAN Zhiqiang, XIE Lingling, CHEN Jiaqi, TANG Hong, WANG Yanbi, ZHAO Caiqin, ZHAO Jiafu. Research Progress on the Interactions of Newcastle Disease Virus M Protein with Host Proteins [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(1): 32-42.
[15]	CHEN Sheng, LIAO Zhihong, XIE Zi, CHEN Feng, XIE Qingmei, SHU Wei. GADD45β Suppresses the Replication of Avian Leukosis Virus Subgroup J in DF-1 Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(6): 1662-1669.

The Effects of Bovine Viral Diarrhoea Virus (BVDV)-induced Ferroptosis on Virus Replication

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 10

References 23

Related Articles 15

Recommended Articles

Metrics

Comments