厚朴总酚对金黄色葡萄球菌的抑菌作用及抗溶血作用

doi:10.11843/j.issn.0366-6964.2018.02.023

摘要/Abstract

摘要：

为了研究厚朴总酚对乳房炎致病菌（主要为金黄色葡萄球菌）的抗菌活性和抗感染机制，首先通过肉汤稀释法检测厚朴总酚对金黄色葡萄球菌的最小抑菌浓度，通过溶血试验、蛋白质免疫印迹试验、荧光定量PCR试验和LDH释放试验从不同层次和角度验证了厚朴总酚对金黄色葡萄球菌致病力的影响。结果显示，厚朴总酚对金黄色葡萄球菌具有一定的抗菌活性，其对金黄色葡萄球菌最小抑菌浓度范围为8～16 μg·mL^-1。另外，本研究结果表明亚抑菌浓度的厚朴总酚可抑制α-溶血素的表达从而降低金黄色葡萄球菌的致病力，并可明显缓解金黄色葡萄球菌α-溶血素介导的细胞的损伤作用。

Abstract:

In order to investigate the antimicrobial activity and anti-infective activity of Total Magnolol on mastitis-related pathogens (mainly Staphylococcus aureus), a broth microdilution method was employed to determine the minimal inhibitory concentrations (MIC) of the Total Magnolol against S. aureus strains. Hemolysis assay, Western blot analysis, real-time RT-PCR assay and cytotoxic assay were further performed to evaluate the influence of this compound on S. aureus virulence. The results showed that the minimal inhibitory concentration of Total Magnolol against S. aureus were 8-16 μg·mL^-1. Furthermore, treatment with Subinhibitory concentration of Total Magnolol could inhibit the expression of α-hemolysin, reduce the pathogenicity of Staphylococcus aureus, and significantly alleviate the injury induced by α-hemolysin of Staphylococcus aureus.

中图分类号:

S853.7

刘桂兰, 周永林, 李文华, 谢玲玲, 刘爱玲, 邓旭明. 厚朴总酚对金黄色葡萄球菌的抑菌作用及抗溶血作用[J]. 畜牧兽医学报, 2018, 49(2): 432-438.

LIU Gui-lan, ZHOU Yong-lin, LI Wen-hua, XIE Ling-ling, LIU Ai-ling, DENG Xu-ming. The Antimicrobial and Anti-hemolysis Effects of Total Magnolol on Staphylococcus aureus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(2): 432-438.

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https://www.xmsyxb.com/CN/Y2018/V49/I2/432

参考文献

[1] 何可云, 董志, 朱毅. 厚朴酚与和厚朴酚的药理学研究现状[J]. 中成药, 2006, 28(5):716-718.
HE K Y, DONG Z, ZHU Y. Research of present situation Honokiol and Honokiol pharmacology[J]. Chinese Traditional Patent Medicine, 2006, 28(5):716-718. (in Chinese)
[2] CHEN H Y, HUNG Y C, LEE E J, et al. The protective efficacy of magnolol in hind limb ischemia-reperfusion injury[J]. Phytomedicine, 2009, 16(10):976-981.
[3] 保志娟, 杨雪琼, 丁中涛, 等. 紫外分光光度法同时测定厚朴酚与和厚朴酚的含量及活性研究[J]. 天然产物研究与开发, 2004, 16(5):435-438.
BAO Z J, YANG X Q, DING Z T, et al. Simultaneous determination of magnolol and honokiol by UV spectrophotometry and study on free radical scavenger activity[J]. Natural Product Research and Development, 2004, 16(5):435-438. (in Chinese)
[4] 王承南, 夏传格. 厚朴药理作用及综合利用研究进展[J]. 经济林研究, 2003, 21(3):80-81.
WANG C N, XIA C G. Research development of Magnolia officinalis medication effect and multiple utilization[J]. Economic Forest Researches, 2003, 21(3):80-81. (in Chinese)
[5] MYLLYS V, ASPLUND K, BROFELDT E, et al. Bovine mastitis in Finland in 1988 and 1995-changes in prevalence and antimicrobial resistance[J]. Acta Vet Scand, 1998, 39(1):119-126.
[6] LEVISON L J, MILLER-CUSHON E K, TUCKER A L, et al. Incidence rate of pathogen-specific clinical mastitis on conventional and organic Canadian dairy farms[J]. J Dairy Sci, 2016, 99(2):1341-1350.
[7] SHIH H C, WEI Y H, LEE C H, et al. Magnolol alters cytokine response after hemorrhagic shock and increases survival in Subsequent intraabdominal sepsis in rats[J]. Shock, 2003, 20(3):264-268.
[8] 吴维萍. 厚朴提取物对不同病原菌抑菌效果的研究[J]. 中国农村医学杂志, 2008, 6(4):5-11.
WU W P. Magnolia bark extract from the study of different pathogenic bacteria antibacterial effect[J]. Chinese Journal of Rural Medicine, 2008, 6(4):5-11. (in Chinese)
[9] RAGLE B E, WARDENBURG J B. Anti-alpha-hemolysin monoclonal antibodies mediate protection against Staphylococcus aureus pneumonia[J]. Infect Immun, 2009, 77(7):2712-2718.
[10] SMITH-PALMER A, STEWART J, FYFE L. Influence of subinhibitory concentrations of plant essential oils on the production of enterotoxins A and B and alpha-toxin by Staphylococcus aureus[J]. J Med Microbiol, 2004, 53(Pt 10):1023-1027.
[11] QIU J Z, FENG H H, LU J, et al. Eugenol reduces the expression of virulence-related exoproteins in Staphylococcus aureus[J]. Appl Environ Microbiol, 2010, 76(17):5846-5851.
[12] 张彬, 薛明, 崔颖, 等. 新兽药消炎醌对乳牛乳腺炎临床治疗试验[J]. 中国兽医科技, 2000, 30(8):28-29.
ZHANG B, XUE M, CUI Y, et al. New veterinary decrease inflammation quinones medicine antiphlogistic quinone of dairy cow mastitis treatment of clinical trials[J]. Chinese Journal of Veterinary Science and Technology, 2000, 30(8):28-29. (in Chinese)
[13] 路建彪, 刘文强. 奶牛乳房炎病原菌的分离与鉴定[J]. 中国兽医杂志, 2007, 43(1):62-63.
LU J B, LIU W Q. Pathogen isolation and identification of dairy cow mastitis[J]. Chinese Journal of Veterinary Medicine, 2007, 43(1):62-63. (in Chinese)
[14] CHAFFER M, LEITNER G, WINKLER M, et al. Coagulase-negative staphylococci and mammary gland infections in cows[J]. J Vet Med B, 1999, 46(10):707-712.
[15] 王志强, 宓伟, 刘现兵, 等. 厚朴体外抑菌作用研究[J]. 时珍国医国药, 2007, 18(11):2763.
WANG Z Q, MI W, LIU X B, et al. The in vitro growth-inhibitory effect of Magnolia officinalis rehd. et wils. (MOR) on bacteria[J]. Lishizhen Medicine and Materia Medica Research, 2007, 18(11):2763. (in Chinese)
[16] BASELGA R, ALBIZU I, DE LA CRUZ M, et al. Phase variation of slime production in Staphylococcus aureus:implications in colonization and virulence[J]. Infect Immun, 1993, 61(11):4857-4862.
[17] WANG J P, RAUNG S L, CHEN C C, et al. The inhibitory effect of magnolol on cutaneous permeability in mice is probably mediated by a nonselective vascular hyporeactivity to mediators[J]. Naunyn Schmiedebergs Arch Pharmacol, 1993, 348(6):663-669.
[18] 寇芮. 动物源耐甲氧西林金黄色葡萄球菌耐药性分析及分子流行病学研究[D]. 南京:南京农业大学, 2010.
KOU R. Study on antibiotic resistance analysis and molecular epidemiology of methicillin-resistant Staphylococcus aureus from animals[D]. Nanjing:Nanjing Agricultural University, 2010. (in Chinese)
[19] 胡付品, 朱德妹, 汪复, 等. 2013年中国CHINET细菌耐药性监测[J]. 中国感染与化疗杂志, 2014, 14(5):369-378.
HU F P, ZHU D M, WANG F, et al. CHINET 2013 surveillance of bacterial resistance in China[J]. Chinese Journal of Infection and Chemotherapy, 2014, 14(5):369-378. (in Chinese)
[20] SAITO J, SAKAI Y, NAGASE H. In vitro anti-mutagenic effect of magnolol against direct and indirect mutagens[J]. Mutat Res, 2006, 609(1):68-73.
[21] LI N, SONG Y, ZHANG W Z, et al. Evaluation of the in vitro and in vivo genotoxicity of magnolia bark extract[J]. Regul Toxicol Pharmacol, 2007, 49(3):154-159.
[22] SAITO J, SHIBUYA K, NAGASE H, et al. Anti-clastogenic effect of magnolol on benzo(a)pyrene-induced clastogenicity in mice[J]. Food Chem Toxicol, 2008, 46(2):694-700.
[23] WANG J, ZHOU X, LIU S, et al. Morin hydrate attenuates Staphylococcus aureus virulence by inhibiting the self-assembly of α-hemolysin[J]. J Appl Microbiol, 2015, 118(3):753-763.
[24] 陈勇川, 谢林利, 熊丽蓉, 等. 黄芩苷/黄芩素对耐甲氧西林金黄色葡萄球菌抗药性的逆转作用研究[J]. 中国药房, 2008, 19(9):644-646.
CHEN Y C, XIE L L, XIONG L R, et al. Reverse effect of baicalin/baicalein on antibiotic resistance of methicillin-resistant Staphylococcus aureus[J]. China Pharmacy, 2008, 19(9):644-646. (in Chinese)
[25] BERUBE B J, WARDENBURG J B. Staphylococcus aureus α-toxin:nearly a century of intrigue[J]. Toxins, 2013, 5(6):1140-1166.
[26] QIU J Z, NIU X D, DONG J, et al. Baicalin protects mice from Staphylococcus aureus pneumonia via inhibition of the cytolytic activity of α-hemolysin[J]. J Infect Dis, 2012, 206(2):292-301.
[27] PAPA R, ARTINI M, CELLINI A, et al. A new anti-infective strategy to reduce the spreading of antibiotic resistance by the action on adhesion-mediated virulence factors in Staphylococcus aureus[J]. Microb Pathog, 2013, 63:44-53.
[28] IKEMOTO S, SUGIMURA K, YOSHIDA N, et al. Antitumor effects of Scutellariae radix and its components baicalein, baicalin, and wogonin on bladder cancer cell lines[J]. Urology, 2000, 55(6):951-955.
[29] SUGA H, SMITH K M. Molecular mechanisms of bacterial quorum sensing as a new drug target[J]. Curr Opin Chem Biol, 2003, 7(5):586-591.
[30] BARON C. A novel strategy to target bacterial virulence[J]. Future Microbiol, 2013, 8(1):1-3.
[31] IWAO Y, TAKANO T, HIGUCHI W, et al. A new staphylococcal cassette chromosome mec IV encoding a novel cell-wall-anchored surface protein in a major ST8 community-acquired methicillin-resistant Staphylococcus aureus clone in Japan[J]. J Infect Chemother, 2012, 18(1):96-104.