牛种布鲁氏菌Ⅳ型分泌系统对巨噬细胞内质网应激和细胞凋亡的影响

doi:10.11843/j.issn.0366-6964.2022.04.019

Abstract

Abstract: In order to study the effect of Brucella bovis type IV secretion system (T4SS) on the endoplasmic reticulum stress and apoptosis of macrophages, the molecular pathogenic mechanism of Brucella was deeply explored. In this study, bovine Brucella vaccine strain A19 and its T4SS promoter deletion strain A19ΔVirB were used to infect mouse macrophages. RNA and total protein were collected at 6, 12 and 24 hours after infection, transcription level and protein expression level of endoplasmic reticulum stress marker molecules GRP78 and CHOP, apoptosis related molecules BAX and BCL-2 were detected by qRT-PCR and Western blot, apoptosis at different time points was detected by flow cytometry. The results showed that at 6, 12 and 24 h after infection, the transcription level and protein expression level of GRP78 in deletion strain A19ΔVirB group were significantly lower than those in parent A19 group (P<0.05); at 6 h after infection, the transcription level and protein expression level of CHOP in the deletion strain group were also significantly lower than those in the parent strain (P<0.05), but at 24 h, it was significantly higher than that of the parent strain (P<0.05); at 24 h after infection,transcription level and protein expression level of BAX in deletion strain A19ΔVirB group were significantly higher than those in parental strains (P<0.05); at 24 h after infection,transcription level and protein expression level of BCL-2 in deletion strain A19ΔVirB group were significantly lower than those in parental strain (P<0.05); the results of flow cytometry further showed that the apoptosis rate of the deletion strain group was significantly higher than that of the parent strain (P<0.05). The results showed that deletion of T4SS would weaken the ability of Brucella bovis to induce endoplasmic reticulum stress and enhance its ability to induce apoptosis.This study laid a foundation for further understanding the pathogenic mechanism of Brucella, and also provided a scientific basis for the functional research of Brucella T4SS in the future.

Key words: Brucella, deletion strain, T4SS, endoplasmic reticulum stress, apoptosis

CLC Number:

YANG Qin, DENG Xiaoyu, XIE Shanshan, YI Jihai, WANG Yong, ZHANG Qian, WANG Zhen, CHEN Chuangfu. Effects of Brucella bovis Type IV Secretion System on Endoplasmic Reticulum Stress and Apoptosis of Macrophages[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(4): 1192-1200.

References 25

[1]	GELETU U S, USMAEL M A, MUMMED Y Y. Seroprevalence and risk factors of small ruminant brucellosis in West Hararghe zone of oromia regional state, Eastern Ethiopia[J]. Vet Med Int, 2021, 2021:6671554.
[2]	耿慧娟. 布鲁氏杆菌在巨噬细胞内可以长期存在的原因[J]. 中西医结合心血管病电子杂志, 2020, 8(28):58-59.GENG H J. Reasons why Brucella can long-term persist within macrophages[J]. Cardiovascular Disease Journal of Integrated Traditional Chinese and Western Medicine (Electronic), 2020, 8(28):58-59. (in Chinese)
[3]	JIAO H W, ZHOU Z X, LI B W, et al. The mechanism of facultative intracellular parasitism of Brucella [J]. Int J Mol Sci, 2021, 22(7):3673.
[4]	KE Y H, WANG Y F, LI W F, et al. Type IV secretion system of Brucella spp. and its effectors[J]. Front Cell Infect Microbiol, 2015, 5:72.
[5]	CELLI J. The intracellular life cycle of Brucella spp. [J]. Microbiol Spectr, 2019, 7(2):10.
[6]	MA Z C, LI R R, HU R R, et al. Brucella abortus BspJ is a nucleomodulin that inhibits macrophage apoptosis and promotes intracellular survival of Brucella [J]. Front Microbiol, 2020, 11:599205.
[7]	何作萍. 羊种布鲁氏菌外膜蛋白诱导的细胞凋亡分子机制[D]. 广州: 南方医科大学, 2016.HE Z P. Molecular mechanism of apoptosis induced by outer membrane proteins of Brucella melitensis [D]. Guangzhou: Southern Medical University, 2016. (in Chinese)
[8]	岳瑞超. MAP诱导内质网应激介导的巨噬细胞凋亡及其对小鼠感染M. bovis 的影响[D]. 北京: 中国农业大学, 2018.YUE R C. MAP induces macrophage apoptosis mediated by ERs and its effect on M. bovis -infected mice[D]. Beijing: China Agriculture University, 2018. (in Chinese)
[9]	孟鹭, 赵冬冬, 吴雨桐, 等. 内质网应激介导的胰岛β细胞凋亡研究进展[J]. 动物医学进展, 2019, 40(8):98-102.MENG L, ZHAO D D, WU Y T, et al. Progress on islet β cells apoptosis mediated by endoplasmic reticulum stress[J]. Progress in Veterinary Medicine, 2019, 40(8):98-102. (in Chinese)
[10]	刘宇婷, 李国新, 王斌. 疱疹病毒与内质网应激[J]. 生物工程学报, 2021, 37(1):67-77.LIU Y T, LI G X, WANG B. Herpesvirus and endoplasmic reticulum stress[J]. Chinese Journal of Biotechnology, 2021, 37(1):67-77. (in Chinese)
[11]	李铭, 邹颖, 郭寒, 等. 钙离子在内质网应激中的作用[J]. 动物医学进展, 2018, 39(9):112-116.LI M, ZOU Y, GUO H, et al. Effect of calcium ion on endoplasmic reticulum stress[J]. Progress in Veterinary Medicine, 2018, 39(9):112-116. (in Chinese)
[12]	ZHI F J, ZHOU D, BAI F R, et al. VceC Mediated IRE1 pathway and inhibited CHOP-induced apoptosis to support Brucella replication in goat trophoblast cells[J]. Int J Mol Sci, 2019, 20(17):4104.
[13]	尹彦龙, 李杨, 白芙蓉, 等. 布鲁氏菌对RAW264. 7细胞内质网应激与细胞因子分泌的影响[J]. 微生物学通报, 2017, 44(12):2896-2904.YIN Y L, LI Y, BAI F Y, et al. Effect of Brucella suis S2 on endoplasmic reticulum stress and cytokines in RAW264. 7 cells[J]. Microbiology China, 2017, 44(12):2896-2904(in Chinese)
[14]	史静雪. 布鲁氏菌分泌蛋白VceA和VceC对细胞自噬和凋亡的影响[D]. 石河子: 石河子大学, 2017.SHI J X. Secretion proteins VceA and VceC of Brucella effects on autophagy and apoptosis[D]. Shihezi: Shihezi University, 2017. (in Chinese)
[15]	相彩霞. 布鲁氏菌VceC蛋白对山羊滋养层细胞内质网应激反应和性腺激素分泌的影响[D]. 杨凌: 西北农林科技大学, 2017.XIANG C X. The endoplasmic reticulum stress and gonadal hormone secretory influence of Brucella VceC on goat trophoblast cells[D]. Yangling: Northwest A & Forestry University, 2017. (in Chinese)
[16]	邓肖玉, 何金科, 杨琴, 等. 牛种布鲁氏菌A19 VirB启动子缺失株的构建及其生物学特性研究[J]. 中国畜牧兽医, 2020, 47(3):864-872.DENG X Y, HE J K, YANG Q, et al. Construction of Brucella abortus A19 VirB promoter deletion strain and study on its biological characteristics[J]. China Animal Husbandry & Veterinary Medicine, 2020, 47(3):864-872. (in Chinese)
[17]	席静，王月丽，邓肖玉，等.STA6介导的巨噬细胞极化对布鲁氏菌胞内存活的影响[J].畜牧兽医学报，2022, 53(1): 263-271.XI J, WANG Y L, DENG X Y, et al. STA6 mediated macrophage polarization on intracellular survival of Brucella [J]. Acta Veternaria et Zootechnica Sinica, 2022, 53(1): 263-271.(in Chinese)
[18]	WANG X G, LIN P F, LI Y, et al. Brucella suis vaccine strain 2 induces endoplasmic reticulum stress that affects intracellular replication in goat trophoblast cells in vitro [J]. Front Cell Infect Microbiol, 2016, 6:19.
[19]	吴同垒, 单晓枫, 康元环, 等. 布鲁菌Ⅳ型分泌系统效应蛋白的研究进展[J]. 中国兽医学报, 2018, 38(3):602-608.WU T L, SHAN X F, KANG Y H, et al. Research advances on type Ⅳ secretion system effectors of Bruella [J]. Chinese Journal of Veterinary Science, 2018, 38(3):602-608. (in Chinese)
[20]	LI P, TIAN M X, BAO Y Q, et al. Brucella rough mutant induce macrophage death via activating IRE1α pathway of endoplasmic reticulum stress by enhanced T4SS secretion[J]. Front Cell Infect Microbiol, 2017, 7:422.
[21]	刘同君, 赵盼, 赵敏孟, 等. 内质网应激标记基因Grp 78参与鹅肥肝免疫/炎症状态的调控[J]. 畜牧兽医学报, 2019, 50(4):727-737.LIU T J, ZHAO P, ZHAO M M, et al. Endoplasmic reticulum stress marker gene Grp 78 is involved in regulation of immune/inflammatory state of goose fatty liver[J]. Acta Veterinaria et Zootechnica Sinica, 2019, 50(4):727-737. (in Chinese)
[22]	胡海, 田明星, 丁婵. CHOP调控内质网应激介导细胞凋亡的机制[J]. 中国预防兽医学报, 2019, 41(2):219.HU H, TIAN M X, DING C. Mechanisms by which CHOP regulates ER stress-mediated apoptosis[J]. Chinese Journal of Preventive Veterinary Medicine, 2019, 41(2):219. (in Chinese)
[23]	支飞杰. 布鲁氏菌GntR缺失菌株构建及其对山羊肺泡巨噬细胞内质网应激的影响[D]. 杨凌: 西北农林科技大学, 2018.ZHI F J. Construction of Brucella GntR gene mutant strain and its effect on endoplasmic reticulum stress in goat alveolar macrophages[D]. Yangling: Northwest A & F University, 2018. (in Chinese)
[24]	弓晶, 解新科. 内质网应激和细胞自噬对肝细胞凋亡的影响[J]. 临床肝胆病杂志, 2019, 35(12):2828-2832.GONG J, XIE X K. Effect of endoplasmic reticulum stress and autophagy on hepatocyte apoptosis[J]. Journal of Clinical Hepatology, 2019, 35(12):2828-2832. (in Chinese)
[25]	高文暄. 微生物感染与细胞凋亡相互作用研究进展[J]. 生物学教学, 2018, 43(11):4-6.GAO W X. Progress in the interaction between microbial infection and apoptosis[J]. Biology Teaching, 2018, 43(11):4-6. (in Chinese)

Effects of Brucella bovis Type IV Secretion System on Endoplasmic Reticulum Stress and Apoptosis of Macrophages

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