鲍曼不动杆菌荚膜厚度对鼠巨噬细胞炎症复合体活化的影响

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江苏大学学报（医学版） ›› 2019, Vol. 29 ›› Issue (02) : 133.

检验医学

鲍曼不动杆菌荚膜厚度对鼠巨噬细胞炎症复合体活化的影响

吴亮1, 陆娟1,2, 王廷廷1, 万晟霞3, 邹治情1, 戴晓玥1,夏雯1, 阴晴4, 陈盛霞1, 许化溪1

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Activation of inflammasome in infection by Acinetobacter baumannii with different capsule thickness

WU Liang1, LU Juan1,2, WANG Ting-ting1, WAN Sheng-xia3, ZOU Zhi-qing1,DAI Xiao-yue1, XIA Wen1, YIN Qing3, CHEN Sheng-xia1, XU Hua-xi1

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摘要

目的: 探讨荚膜厚度不同的鲍曼不动杆菌激活鼠巨噬细胞Raw 264.7炎症复合体能力的差异。方法: 采用刚果红染色法观察鲍曼不动杆菌荚膜,筛选出荚膜厚度差异显著的两株菌株。将两株菌分别感染Raw 264.7,以实时定量 PCR法分析感染6 h的细胞NOD样受体家族3(NODlike receptors, NLRP3)、Caspase1和IL1β基因表达量;流式细胞术分析感染1、3和6 h的细胞活性氧表达量。结果：刚果红染色可以清晰地观察到鲍曼不动杆菌周围一圈不着色的荚膜,选择出荚膜厚度差异显著的两株鲍曼不动杆菌用于感染细胞。不同荚膜厚度的鲍曼不动杆菌感染均可刺激Raw 264.7细胞NLRP3、Caspase1和IL1β mRNA表达上调,其中厚荚膜菌株诱导细胞NLRP3和IL1β mRNA表达能力显著高于薄荚膜菌株（P<0.05）;不同荚膜厚度的鲍曼不动杆菌感染3 h和6 h时,细胞活性氧表达量均显著高于对照组(P<0.05),且厚荚膜菌株诱导活性氧表达能力显著高于薄荚膜菌株（P<0.05）。>

Abstract

Objective: To investigate the activation of Raw 264.7 cell inflammasome by Acinetobacter baumannii(A. baumannii) with different capsule thickness. Methods: A. baumannii capsule was stained by Congo red, and selected two strains with significant differences in capsule thickness. The Raw 264.7 cells were infected with two strains of bacteria, and the cells culture was collected in 6 h. The gene expression of NODlike receptors(NLRP3), caspase1 and IL1β were determined by realtime PCR. The level of reactive oxygen species(ROS) in Raw 264.7 cells was determined by flow cytometry in 1 h, 3 h and 6 h. Results: The capsule around A. baumannii could not be stained by Congo red, and it was easy to be observed. The expression of NLRP3, caspase1 and IL1β mRNA in Raw 264.7 cells infected by two strains of A. baumannii were increased significantly compared with normal cell control. The expression of NLRP3 and IL1β in thick capsular strain was significantly higher than that of the thin capsular strain（P<0.05）, and there was no significant difference in caspase1 expression between the two strains. Both the two strains could upregulate the ROS production of Raw 264.7 cells. The ROS production in two strains infection group was significantly higher than that of normal cells control in 3 h and 6 h（P<0.05）, and the ROS production in the thick capsular strain group was significantly higher than that of the thin capsular strain group（P<0.05）. conclusion:="" a.="" baumannii="" with="" different="" capsule="" thickness="" showed="" different="" activation="" abilities="" of="" inflammasome="" in="" raw="" 264.7="" cell="" and="" the="" activation="" ability="" of="" inflammasome="" induced="" by="" the="" thick="" capsule="" strain="" was="" stronger.="">

关键词

鲍曼不动杆菌 / 荚膜 / 炎症复合体 / 活性氧

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吴亮1, 陆娟1,2, 王廷廷1, 万晟霞3, 邹治情1, 戴晓玥1,夏雯1, 阴晴4, 陈盛霞1, 许化溪1. 鲍曼不动杆菌荚膜厚度对鼠巨噬细胞炎症复合体活化的影响[J]. 江苏大学学报（医学版）, 2019, 29(02): 133

WU Liang1, LU Juan1,2, WANG Ting-ting1, WAN Sheng-xia3, ZOU Zhi-qing1,DAI Xiao-yue1, XIA Wen1, YIN Qing3, CHEN Sheng-xia1, XU Hua-xi1. Activation of inflammasome in infection by Acinetobacter baumannii with different capsule thickness [J]. Journal of Jiangsu University(Medicine Edition), 2019, 29(02): 133

参考文献

［1］Nowak P, Paluchowska P. Acinetobacter baumannii: biology and drug resistancerole of carbapenemases［J］. Folia Histochem Cytobiol, 2016, 54(2):61-74.

［2］Ko KS. Antibioticresistant clones in Gramnegative pathogens: presence of global clones in Korea［J］. J Microbiol, 2019, 75(3):195-202.

［3］Eze EC, Chenia HY, El Zowalaty ME. Acinetobacter baumannii biofilms: effects of physicochemical factors, virulence, antibiotic resistance determinants, gene regulation, and future antimicrobial treatments［J］. Infect Drug Resist, 2018,11: 2277-2299.

［4］Wood CR, Ohneck EJ, Edelmann RE, et al. A lightregulated type Ⅰ pilus contributes to Acinetobacter baumannii biofilm, motility, and virulence functions［J］. Infect Immun, 2018, 86(9):e00442.

［5］刘倩倩, 刘运德, 张琼, 等. Syk调控单增李斯特菌感染中炎症复合体的活化［J］. 天津医药, 2014, 42(5):432-435.

［6］王怀兵, 牛凤梅, 端木素丽. 细菌荚膜染色的改良［J］. 黔南民族医专学报, 2002, 16(4):206.

［7］何发明, 范晶, 余泽波, 等. ICU中痰标本来源的鲍曼不动杆菌的临床意义分析［J］. 中国抗生素杂志, 2012, 37(5):357-361.

［8］刘华, 黄学斌, 肖代文, 等. 重症监护病房鲍曼不动杆菌感染／定植情况及耐药性分析［J］. 实用医院临床杂志, 2012, 9(6):102-104.

［9］Jacobs AC, Thompson MG, Black CC, et al. AB5075, a highly virulent isolate of Acinetobacter baumannii, as a model strain for the evaluation of pathogenesis and antimicrobial treatments［J］. MBio, 2014, 5(3):e01076-14.

［10］Lee CR, Lee JH, Park M, et al. Biology of Acinetobacter baumannii: pathogenesis, antibiotic resistance mechanisms, and prospective treatment options［J］. Front Cell Infect Microbiol, 2017, 7:55.

［11］Zhou K, Tang X, Wang L, et al. An emerging clone (ST457) of Acinetobacter baumannii clonal complex 92 with enhanced virulence and increasing endemicity in south China［J］. Clin Infect Dis, 2018, 67(2): S179-S188.

［12］孙敬, 陈会, 余理智, 等. 痰液培养定植菌与病原菌判断方法的探讨［J］. 江西医学检验, 2006, 24(6):485-488.

［13］ParedesJuarez GA, de Haan BJ, Faas MM, et al. The role of pathogenassociated molecular patterns in inflammatory responses against alginate based microcapsules［J］. J Control Release, 2013, 172(3):983-992.

［14］Evrard B, Balestrino D, Dosgilbert A, et al. Roles of capsule and lipopolysaccharide O antigen in interactions of human monocytederived dendritic cells and Klebsiella pneumoniae［J］. Infect Immun, 2010, 78(1):210-219.

［15］Franchi L, Mcdonald C, Kanneganti TD, et al. Nucleotidebinding oligomerization domainlike receptors: intracellular pattern recognition molecules for pathogen detection and host defense［J］. J Immunol, 2006, 177 (6):3507-3513.

［16］Martinon F, Petrilli V, Mayor A, et al. Goutassociated uric acid crystals activate the NALP3 inflammasome［J］. Nature, 2006, 440(7081):237-241.

［17］Zhou R, Yazdi AS, Menu P, et al. A role for mitochondria in NLRP3 inflammasome activation［J］. Nature, 2011, 469(7329):221-225.

［18］Kang MJ, Jo SG, Kim DJ, et al. NLRP3 inflammasome mediates interleukin1β production in immune cells in response to Acinetobacter baumannii and contributes to pulmonary inflammation in mice［J］. Immunology, 2017, 150(4):495-505.

［19］Castronovo G, Clemente AM, Antonelli A, et al. Differences in inflammatory response induced by two representatives of clades of the Pandemic ST258 Klebsiella pneumoniae clonal lineage producing KPCtype carbapenemases［J］. PLoS One, 2017, 12(1):e170125.

［20］Feng H, Gu J, Gou F, et al. High glucose and lipopolysaccharide prime NLRP3 inflammasome via ROS／TXNIP pathway in mesangial cells［J］. J Diabetes Res, 2016, 2016:6973175.

［21］Sorbara MT, Girardin SE. Mitochondrial ROS fuel the inflammasome［J］. Cell Res, 2011, 21(4):558-560.