致病性大肠杆菌HPI对Caspase-1细胞焦亡相关分子表达的影响

doi:10.16431/j.cnki.1671-7236.2019.11.006

Abstract

Abstract: In order to study the molecular mechanism of cell death in porcine macrophages induced by pathogenic E.coli HPI from Saba pig in Yunnan,the pig derived macrophages infected with HPI positive strain of Saba pig in Yunnan were taken as the breakthrough point. E.coli was isolated from feces of yellow-white dysentery in a large scale farm in Chuxiong,Yunnan.The HPI irp2 gene was detected by PCR technology.Macrophages were infected with HPI positive strains (HPI⁺) and negative strains (HPI^-),respectively,and LPS+ATP group and blank control group were set up,cells and their supernatants were collected at 0.5 and 6 h.The expression levels of Caspase-1,IL-1β and IL-18 in different groups were detected by Real-time PCR,and the contents of pro-IL-1β,pro-IL-18,IL-1β and IL-18 in cell supernatant were detected by ELISA.The results showed that pathogenic E.coli was successfully isolated and HPI irp2 gene positive strains were successfully obtained by PCR.Real-time PCR results showed that the expression levels of Caspase-1,IL-1β and IL-18 of HPI⁺ and HPI^- groups were all higher than those of blank control group.It showed an upward trend,and HPI⁺ group was higher than HPI^- group.ELISA results showed that compared with blank control group,the expression of pro-IL-1β,pro-IL-18,IL-1β and IL-18 in HPI⁺ and HPI^- groups were generally up-regulated,and the HPI⁺ group was higher than that in HPI^- group.The results showed that the pathogenic E.coli HPI of Saba pig in Yunnan could promote the release of inflammatory cytokines IL-1β and IL-18 from pig macrophages by up regulating the expression of Caspase-1,IL-1β and IL-18 mRNA in porcine macrophages,and eventually induce inflammation.

Key words: pathogenic E.coli; HPI; macrophages; cell death

CLC Number:

S852.61

CHA Jinlong, LIU Chaoying, GAO Hong, WANG Shiyu, SHAN Chunlan, JING Linxi, WANG Hao, YANG Wei. Effect of Pathogenic E.coli HPI on the Expression of Burnout-related Molecules in Caspase-1 Cells[J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(11): 3181-3189.

References

[1] MOKRACKA J,KOCZURA R,KAZNOWSKI A.Yersiniabactin and other siderophores produced by clinical isolates of Enterobacter spp. and Citrobacter spp.[J].FEMS Immunology and Medical Microbiology,2004,40(1):51-55.
[2] MAGISTRO G,MAGISTRO C,STIEF C G,et al.The high-pathogenicity island (HPI) promotes flagellum-mediated motility in extraintestinal pathogenic Escherichia coli[J].PLoS One,2017,12(10):e0183950.
[3] 王艳萍,郭时金,徐倩倩,等.禽致病性大肠杆菌HPI毒力岛irp2蛋白表达、纯化及其间接ELISA方法建立[J].中国兽医学报,2018,38(2):287-292. WANG Y P,GUO S J,XU Q Q,et al.Establishment of indirect ELISA method and avian pathogenic E.coli HPI irp2 protein expression and purification[J]. Chinese Journal of Veterinary Science,2018,38(2):287-292.(in Chinese)
[4] 史秋梅,张艳英,房海,等.腹泻水貂检出携带耶尔森菌HPI毒力岛的大肠杆菌[J].中国兽医学报,2012,32(6):857-861. SHI Q M,ZHANG Y Y,FANG H,et al.Detection of pathogenicity islands of Yersinia enterocolitica HPI from Escherichia coli strains in diarrheic minks[J].Chinese Journal of Veterinary Science,2012,32(6):857-861.(in Chinese)
[5] 张艳英,史秋梅,房海,等.腹泻仔貉检出携带耶尔森菌HPI毒力岛的大肠杆菌[J].中国人兽共患病学报,2012,28(2):179-182. ZHANG Y Y,SHI Q M,FANG H,et al.Detection on pathogenicity islands of Yersinia enterocolitica HPI from Escherichia coli strains in diarrheic young raccoon dogs[J].Chinese Journal of Zoonoses,2012,28(2):179-182.(in Chinese)
[6] 魏红霞.ATP通过调节AMPK信号诱导巨噬细胞炎症小体活化和细胞焦亡[D].广州:暨南大学,2017. WEI H X.ATP-induced macrophage inflammasome activation and pyroptosis is regulated by AMP-activated protein kinase in macrophages[D].Guangzhou:Jinan University,2017.(in Chinese)
[7] 邬皓晨,陈勇军,徐易尘,等.Caspase-1在炎症及程序性细胞死亡过程中的作用[J].中国细胞生物学学报,2011,33(2):182-189. WU H C,CHEN Y J,XU Y C,et al.Role of Caspase-1 in inflammation and programmed cell death[J].Chinese Journal of Cell Biology,2011,33(2):182-189.(in Chinese)
[8] 徐亚芳,陈创夫,王浩,等.AIR对布鲁氏菌感染引发的炎症反应的影响[J].中国畜牧兽医,2016,43(6):1437-1445. XU Y F,CHEN C F,WANG H,et al.Effect of AIR on inflammatory reaction infected by Brucella[J].China Animal Husbandry & Veterinary Medicine,2016,43(6):1437-1445.(in Chinese)
[9] 游宇来,王涛,龚建平.细胞焦亡与感染性疾病研究进展[J].检验医学与临床,2018,24:3792-3795. YOU Y L,WANG T,GONG J P.Advances in research on cell burn-out and infectious diseases[J].Laboratory Medicine and Clinic,2018,24:3792-3795.(in Chinese)
[10] 崔国东,张雪梅,陈立杰.IL-1β在急性脑缺血中作用机制的研究进展[J].临床神经病学杂志,2015,28(6):466-468. CUI G D,ZHANG X M,CHEN L J.Research progress on the mechanism of IL-1 beta in acute cerebral ischemia[J].Journal of Clinical Neurology,2015,28(6):466-468.(in Chinese)
[11] NASI M,PECORINI S,DE BIASI S,et al.Altered expression of PYCARD,IL-1β,IL-18 and NAIP in successfully treated HIV⁺ patients with low CD4/CD8 ratio[J].Journal of Infectious Diseases,2019,219(11):1743-1748.
[12] 李海花,张蕾,杨春蕾,等.PCV2通过NF-κB/NLRP3信号通路调控体外培养PAMs分泌IL-1β[J].中国畜牧兽医,2016,43(9):2366-2372. LI H H,ZHANG L,YANG C L,et al.PCV2 regulates PAMs secreting IL-1β through the NF-κB/NLRP3 signalling pathway in vitro[J].China Animal Husbandry & Veterinary Medicine,2016,43(9):2366-2372.(in Chinese)
[13] 林静,李大主.细胞焦亡:一种新的细胞死亡方式[J].国际免疫学杂志,2011,34(3):213-216. LIN J,LI D Z.Pyroptosis:A caspase-1-dependent cell death[J].International Journal of Immunology,2011,34(3):213-216.(in Chinese)
[14] 刘蓓桦,候梁,罗伟珏,等.LPS与ATP共同诱导巨噬细胞中NLRP3炎症小体的激活[J].北京农学院学报,2018,33(1):74-78. LIU B H,HOU L,LUO W J,et al.Mechanisms of lipopolysaccharide induced the activation of NLRP3 inflammation immune response in macrophages[J].Journal of Beijing University of Agriculture,2018,33(1):74-78.(in Chinese)
[15] 蒋建烨.NLRP3炎症复合体信号通路分子在幽门螺杆菌感染小鼠模型中表达的研究[D].衡阳:南华大学,2012. JIANG J Y.Study of expression of NLRP3 inflammasomes in the mice of Helicobacter pylori infection[D].Hengyang:University of South China,2012.(in Chinese)
[16] 梁文杰,马雪莲,王萱,等.化瘀解毒方对梗阻性肾病大鼠肾脏NLRP3、Caspase1、IL-1β表达的影响[J].中国中西医结合杂志,2017,37(4):470-475. LIANG W J,MA X L,WANG X,et al.Effect of Huayu Jiedu Recipe on the expressions of NLRP3,Caspase1,and IL-1β in kidneys of obstructive nephropathy rats[J].Chinese Journal of Integrated Traditional and Western Medicine,2017,37(4):470-475.(in Chinese)
[17] HUGHES M M,O'NEILL L.Metabolic regulation of NLRP3[J].Immunological Reviews,2018,281(1):88-98.
[18] ZHANG Y,LIU X,BAI X,et al.Melatonin prevents endothelial cell pyroptosis via regulation of long noncoding RNA MEG3/miR-223/NLRP3 axis[J].Journal of Pineal Research,2018,64(2):e12449.
[19] HITZLER I,SAYI A,KOHLER E,et al.Caspase-1 has both proinflammatory and regulatory properties in Helicobacter infections,which are differentially mediated by its substrates IL-1β and IL-18[J].Journal of Immunology,2012,188(8):3594-3602.
[20] SAUER J D,WITTE C E,ZEMANSKY J,et al.Listeria monocytogenes triggers AIM2 mediated pyroptosis upon infrequent bacteriolysis in the macrophage cytosol[J].Cell Host Microbe,2010,7(5):412-419
[21] ZEISEL M B,NEFF L A,RANDLE J,et al.Impaired release of IL-18 from fibroblast-like synoviocytes activated with proteinⅠ/Ⅱ,a pathogen-associated molecular pattern from oral Streptococci,results from defective translation of IL-18 mRNA in pro-IL-18[J].Cellular Microbiology,2004,6(6):593-598.
[22] SAUER J D,WITTE C E,ZEMANSKY J,et al.Listeria monocytogenes triggers AIM2-mediated pyroptosis upon infrequent bacteriolysis in the macrophage cytosol[J].Cell Host Microbe,2010,7(5):412-419.

Effect of Pathogenic E.coli HPI on the Expression of Burnout-related Molecules in Caspase-1 Cells

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