香叶木素对嗜水气单胞菌气溶素表达的抑制作用
CSTR:
作者:
作者单位:

中国水产科学研究院长江水产研究所,武汉 430223

作者简介:

李声平, E-mail: lsp19970906@163.com

通讯作者:

董靖, E-mail: dongjing@yfi.ac.cn

中图分类号:

S917.1

基金项目:

国家自然科学基金项目(31702368); 现代农业产业技术体系建设专项 (CARS-46)


Inhibitory effect of diosmetin on aerolysin expression in Aeromonas hydrophila
Author:
Affiliation:

Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, China

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    摘要:

    为筛选出抗嗜水气单胞菌感染的天然化合物,以嗜水气单胞菌和香叶木素为研究对象,通过最小抑菌浓度测定、生长曲线、溶血试验、免疫印迹、荧光定量PCR和细胞毒性试验研究了香叶木素对嗜水气单胞菌气溶素表达的抑制作用。结果发现,对嗜水气单胞菌生长没有影响的香叶木素能在较低浓度下通过抑制气溶素的表达降低共培养物上清液的溶血活性;荧光定量PCR试验发现香叶木素能剂量依赖性地降低气溶素编码基因aerA 的表达。此外,通过活/死细胞染色发现香叶木素可减轻气溶素介导的细胞损伤。以上结果表明,香叶木素通过降低aerA的转录水平降低了嗜水气单胞菌的体外致病力,可作为一种抗嗜水气单胞菌感染的候选药物。

    Abstract:

    To screen out natural compounds against Aeromonas hydrophilaA. hydrophila) infection, minimal inhibitory concentration determination, growth curves, hemolysis, Western blot, qRT-PCR and cytotoxicity assay were performed to assess the inhibitory effect of diosmetin on the expression of aerolysin secreted by A. hydrophila. The results showed that diosmetin without anti- A. hydrophila activity could inhibit the hemolytic activity of bacterial supernatants by decreasing the transcription of aerolysin gene. Moreover, live/dead staining assay showed that diosmetin could attenuate cell injury mediated by aerolysin in A549 cells. These findings demonstrated that diosmetin could reduce the pathogenicity of A. hydrophila by inhibiting the expression of aerolysin and diosmetin could be chosen as a promising candidate drug against A. hydrophila infection.

    表 1 试验中使用的引物Table 1 Primer pairs used in test
    图1 嗜水气单胞菌与香叶木素共培养的生长曲线Fig.1 Growth curves of A. hydrophila co-cultured with diosmetin
    图2 香叶木素对嗜水气单胞菌上清液溶血的抑制作用Fig.2 Impact of diosmetin on hemolytic activity of bacterial supernatants
    图3 不同质量浓度香叶木素处理后培养物上清液中气溶素的含量Fig.3 Amount of aerolysin in bacterial supernatants after a co-incubation with diosmetin at different concentrations
    图4 香叶木素处理后嗜水气单胞菌aerA基因的转录水平Fig.4 Transcription levels of aerA genes in A. hydrophila treated with diosmetin
    图5 香叶木素对气溶素介导的细胞损伤的保护作用Fig.5 Protective effect of diosmetin against aerolysin mediated cell injury
    参考文献
    [1] 蒋长军,管翠翠,胡小敏,等.黄芩苷对嗜水气单胞菌生物膜形成的影响及体内外的抑菌作用[J].微生物学报,2021,61(7):2112-2120.JIANG C J,GUAN C C,HU X M,et al.Bacteriostatic and antibiofilm effects of baicalin on Aeromonas hydrophila:in vitro and in vivo evaluation[J].Acta microbiologica Sinica,2021,61(7):2112-2120(in Chinese with English abstract).
    [2] 吴璇,袁海文,谢晓东,等.斑点叉尾鮰套肠病病原嗜水气单胞菌的分离与鉴定[J].中国畜牧兽医,2017,44(10):3057-3062.WU X,YUAN H W,XIE X D,et al.Isolation and identification of Aeromonas hydrophila from the case of channel catfish bowel disease[J].China animal husbandry & veterinary medicine,2017,44(10):3057-3062(in Chinese with English abstract).
    [3] 薛明洋,周勇,梁宏伟,等.一株中华鳖源嗜水气单胞菌的分离鉴定及毒力基因分析[J].中国农学通报,2021,37(20):152-159.XUE M Y,ZHOU Y,LIANG H W,et al.Isolation,identification and virulence gene analysis of an Aeromonas hydrophila from diseased Pelodiscus sinensis[J].Chinese agricultural science bulletin,2021,37(20):152-159(in Chinese with English abstract).
    [4] 胡安东,张明洋,张飘,等.鲤源嗜水气单胞菌分离鉴定、耐药性及毒力基因检测[J].中国畜牧兽医,2019,46(7):2167-2175.HU A D,ZHANG M Y,ZHANG P,et al.Isolation,identification,drug resistance and virulence genes detection of Aeromonas hydrophila from carp[J].China animal husbandry & veterinary medicine,2019,46(7):2167-2175(in Chinese with English abstract).
    [5] HOSSAIN M J,SUN D W,MCGAREY D J,et al.An Asian origin of virulent Aeromonas hydrophila responsible for disease epidemics in United States-farmed catfish[J/OL].mBio,2014,5(3):e00848-e00814[2022-05-06].https://doi.org/10.1128/mBio.00848-14.
    [6] 张国亮,吕利群.高度耐药嗜水气单胞菌的定向诱导及其交叉耐药性分析[J].淡水渔业,2016,46(6):56-63.ZHANG G L,Lü L Q.Directional induction of antibiotics-resistant Aeromonas hydrophila and analysis of its multiple drug resistance[J].Freshwater fisheries,2016,46(6):56-63(in Chinese with English abstract).
    [7] NOVITA H,RUSMANA I,YUHANA M,et al.Uji in vitro bakteri anti quorum sensing pendegradasi acyl homoserine lactone Aeromonas hydrophila[J].Jurnal riset akuakultur,2017,11(3):291-296.
    [8] LJUNGH ?,WADSTR?M T.Aeromonas toxins[J].Pharmacology & therapeutics,1981,15(3):339-354.
    [9] 杜娜.嗜水气单胞菌耐药性分析及气溶素抗血清的免疫保护性研究[D].武汉:华中农业大学,2013.DU N.Drug-resistance analysis of Aeromonas hydrophila and study on immunoprotecive of antiserum of aerolysin[D].Wuhan:Huazhong Agricultural University,2013(in Chinese with English abstract).
    [10] 刘明智,叶星,田园园,等.嗜水气单胞菌气溶素和溶血素基因的克隆与结构预测[J].江西农业大学学报,2010,32(2):356-362.LIU M Z,YE X,TIAN Y Y,et al.Cloning and structure prediction of aerA gene and hlyA gene of Aeromonas hydrophila[J].Acta Agriculturae Universitatis Jiangxiensis,2010,32(2):356-362(in Chinese with English abstract).
    [11] CHAKRABORTY T,HUHLE B,HOF H,et al.Marker exchange mutagenesis of the aerolysin determinant in Aeromonas hydrophila demonstrates the role of aerolysin in A.hydrophila-associated systemic infections[J].Infection and immunity,1987,55(9):2274-2280.
    [12] 马纳,李亚静,范吉平.香叶木素药理作用研究进展[J].辽宁中医药大学学报,2018,20(9):214-217.MA N,LI Y J,FAN J P.Research progress on pharmacological action of diosmetin[J].Journal of Liaoning University of Traditional Chinese Medicine,2018,20(9):214-217(in Chinese with English abstract).
    [13] CLSI. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically-eighth edition: approved standard M07-A8[M]. Wayne, PA, USA: CLSI, 2009.
    [14] IACOVACHE I,DEGIACOMI M T,PERNOT L,et al.Dual chaperone role of the C-terminal propeptide in folding and oligomerization of the pore-forming toxin aerolysin[J/OL].PLoS pathogens,2011,7(7):e1002135[2022-05-06]. https://pubmed.ncbi.nlm.nih.gov/21779171/.DOI: 10.1371/journal.ppat.1002135.
    [15] 王群,曾高峰,王燕,等.香叶木素在代谢综合征中的作用[J].生命的化学,2021,41(8):1701-1708.WANG Q,ZENG G F,WANG Y,et al.Role of diosmetin in metabolic syndrome[J].Chemistry of life,2021,41(8):1701-1708(in Chinese with English abstract).
    [16] MENG J C,ZHU Q X,TAN R X.New antimicrobial mono- and sesquiterpenes from Soroseris hookeriana subsp. erysimoides[J].Planta Med,2000,66(6):541-544.
    [17] LI H L,WEI Y Y,LI X H,et al.Diosmetin has therapeutic efficacy in colitis regulating gut microbiota,inflammation,and oxidative stress via the circ-Sirt1/Sirt1 axis[J].Acta pharmacologica sinica,2022,43(4):919-932.
    [18] GOMEZ-CHANG E,URIBE-ESTANISLAO G V,MARTINEZ-MARTINEZ M,et al.Anti-Helicobacter pylori potential of three edible plants known as quelites in Mexico[J].Journal of medicinal food,2018,21(11):1150-1157.
    [19] 马亮,董靖,张国栋,等. 克氏原螯虾源普通变形杆菌分离、鉴定及联合药敏试验[J].淡水渔业,2022,52(5):53-59.MA L, DONG J, ZHANG G D, et al. Identification and synergistic susceptibility of Proteus vulgaris isolated from Procambarus clarkia[J]. Freshwater fisheries, 2022,52(5):53-59(in Chinese with English abstract).
    [20] CHAN B C L,IP M,GONG H,et al.Synergistic effects of diosmetin with erythromycin against ABC transporter over-expressed methicillin-resistant Staphylococcus aureus (MRSA) RN4220/pUL5054 and inhibition of MRSA pyruvate kinase[J].Phytomedicine,2013,20(7):611-614.
    [21] SUN B,LUO H Z,JIANG H,et al.Inhibition of quorum sensing and biofilm formation of esculetin on Aeromonas hydrophila[J/OL].Frontiers in microbiology,2021,12:737626[2022-05-06].https://pubmed.ncbi.nlm.nih.gov/34630364/. DOI: 10.3389/fmicb.2021.737626.
    [22] DONG J,ZHANG L S,LIU Y T,et al.Thymol protects channel catfish from Aeromonas hydrophila infection by inhibiting aerolysin expression and biofilm formation[J/OL].Microorganisms,2020,8(5):636[2022-05-06]. https://pubmed.ncbi.nlm.nih.gov/32349419/. DOI:10.3390/microorganisms8050636.
    [23] LIU S,ZHOU X,LI W H,et al.Diosmetin inhibits the expression of alpha-hemolysin in Staphylococcus aureus[J].Antonie van Leeuwenhoek,2015,108(2):383-389.
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李声平,马亮,闫天慧,艾晓辉,董靖.香叶木素对嗜水气单胞菌气溶素表达的抑制作用[J].华中农业大学学报,2022,41(6):217-221

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  • 收稿日期:2022-05-06
  • 在线发布日期: 2022-12-09
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