Download PDF
Planta Med 2013; 79(02): 110-115
DOI: 10.1055/s-0032-1328068
Biological and Pharmacological Activity
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

Silibinin In Vitro Protects A549 Cells from Staphylococcus aureus-Mediated Injury and In Vivo Alleviates the Lung Injury of Staphylococcal Pneumonia

Xin Wang   *
1   College of Quartermaster Technology, Jilin University, Changchun, PR China
,
Jing Dong   *
2   Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin University, Changchun, PR China
,
Xiaohan Dai
2   Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin University, Changchun, PR China
,
Yu Zhang
2   Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin University, Changchun, PR China
,
Jianfeng Wang
2   Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin University, Changchun, PR China
,
Hongen Li
2   Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin University, Changchun, PR China
,
Chongjian Lu
2   Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin University, Changchun, PR China
,
Wei Tan
2   Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin University, Changchun, PR China
,
Xiaohan Gao
1   College of Quartermaster Technology, Jilin University, Changchun, PR China
,
Xuming Deng
2   Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin University, Changchun, PR China
,
Shijin Bu
3   College of Veterinary Medicine, Yangzhou University, Yangzhou, PR China
,
Xiaodi Niu
1   College of Quartermaster Technology, Jilin University, Changchun, PR China
› Author Affiliations
Further Information

Publication History

received 05 July 2012
revised 13 November 2012

accepted 18 November 2012

Publication Date:
08 January 2013 (online)

    Permissions and Reprints

Abstract

In this study, hemolysis, Western blot, and real-time RT-PCR assays were performed to evaluate silibininʼs activity against S. aureus α-toxin secretion. In addition, live/dead cell staining and lactate dehydrogenase activity assays were introduced to examine the influence of silibinin on α-toxin-induced cell injury in human alveolar epithelial cells. Furthermore, we tested the influence of silibinin on S. aureus pneumonia in a mouse model. We show that silibinin inhibits the expression of α-toxin in a dose-dependent manner and alleviates α-toxin-induced lung cell injury. The IC50 of silibinin that inhibits the hemolytic activity of S. aureus was 14.33 µg/mL for strain 8325–4. Moreover, this compound provides effective protection on the lung injury of staphylococcal pneumonia.

Key words

Staphylococcus aureus - α-toxin - silibinin - Silybum marianum - Asteraceae - A549 cells - pneumonia

* Both authors contributed equally to this work.


 

  • References

  • 1 Lowy FD. Staphylococcus aureus infections. N Engl J Med 1998; 339: 520-532
    CrossrefPubMedGoogle Scholar
  • 2 Rubinstein E, Kollef MH, Nathwani D. Pneumonia caused by methicillin-resistant Staphylococcus aureus . Clin Infect Dis 2008; 46: 378-385
    CrossrefPubMedGoogle Scholar
  • 3 Athanassa Z, Siempos II, Falagas ME. Impact of methicillin resistance on mortality in Staphylococcus aureus VAP: a systematic review. Eur Respir J 2008; 31: 625-632
    CrossrefPubMedGoogle Scholar
  • 4 Rasko DA, Sperandio V. Anti-virulence strategies to combat bacteria-mediated disease. Nat Rev Drug Discov 2010; 9: 117-128
    CrossrefPubMedGoogle Scholar
  • 5 Qiu J, Wang D, Xiang H, Feng H, Jiang Y, Xia L, Dong J, Lu J, Yu L, Deng X. Subinhibitory concentrations of thymol reduce enterotoxins A and B and α-hemolysin production in Staphylococcus aureus isolates. PLoS ONE 2010; 5: e9736
    CrossrefPubMedGoogle Scholar
  • 6 Kobayashi SD, DeLeo FR. An update on community-associated MRSA virulence. Curr Opin Pharmacol 2009; 9: 545-551
    CrossrefPubMedGoogle Scholar
  • 7 Rasooly R, Do PM, Friedman M. Inhibition of biological activity of staphylococcal enterotoxin A (SEA) by apple juice and apple polyphenols. J Agric Food Chem 2010; 58: 5421-5426
    CrossrefPubMedGoogle Scholar
  • 8 Lin YC, Schlievert PM, Anderson MJ, Fair CL, Schaefers MM, Muthyala R, Peterson ML. Glycerol monolaurate and dodecylglycerol effects on Staphylococcus aureus and toxic shock syndrome toxin-1 in vitro and in vivo . PLoS One 2009; 9: e7499
    PubMedGoogle Scholar
  • 9 Recsei P, Kreiswirth B, OʼReilly M, Schlievert P, Gruss A, Novick RP. Regulation of exoprotein gene expression in Staphylococcus aureus by agr. Mol Gen Genet 1986; 202: 58-61
    CrossrefPubMedGoogle Scholar
  • 10 Song L, Hobaugh MR, Shustak C, Cheley S, Bayley H, Gouaux JE. Structure of staphylococcal alpha-hemolysin, a heptameric transmembrane pore. Science 1996; 274: 1859-1866
    CrossrefPubMedGoogle Scholar
  • 11 McElroy MC, Harty HR, Hosford GE, Boylan GM, Pittet JF, Foster TJ. Alpha-toxin damages the air–blood barrier of the lung in a rat model of Staphylococcus aureus–induced pneumonia. Infect Immun 1999; 67: 5541-5544
    PubMedGoogle Scholar
  • 12 Bubeck Wardenburg J, Patel RJ, Schneewind O. Surface proteins and exotoxins are required for the pathogenesis of Staphylococcus aureus pneumonia. Infect Immun 2007; 75: 1040-1044
    CrossrefPubMedGoogle Scholar
  • 13 Wellington K, Jarvis B. Silymarin: a review of its clinical properties in the management of hepatic disorders. BioDrugs 2001; 15: 465-489
    CrossrefPubMedGoogle Scholar
  • 14 Singh RP, Deep G, Chittezhath M, Kaur M, Dwyer-Nield LD, Malkinson AM, Agarwal R. Effect of silibinin on the growth and progression of primary lung tumors in mice. J Natl Cancer Inst 2006; 98: 846-855
    CrossrefPubMedGoogle Scholar
  • 15 Agarwal C, Singh RP, Dhanalakshmi S, Tyagi AK, Tecklenburg M, Sclafani RA, Agarwal R. Silibinin up-regulates the expression of cyclin-dependent kinase inhibitors and causes cell cycle arrest and apoptosis in human colon carcinoma HT-29 cells. Oncogene 2003; 22: 8271-8282
    CrossrefPubMedGoogle Scholar
  • 16 Flaig TW, Gustafson DL, Su LJ, Zirrolli JA, Crighton F, Harrison GS, Pierson AS, Agarwal R, Glodé LM. A phaseI pharmacokinetic study of silybin-phytosome in prostate cancer patients. Invest New Drugs 2007; 25: 139-146
    CrossrefPubMedGoogle Scholar
  • 17 Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing; Fifteenth informational supplement. CL SI/NCCLS document M100-S15. Wayne, PA: Clinical and Laboratory Standards Institute (CLSI); 2005
    Google Scholar
  • 18 Qiu J, Wang J, Luo H, Du X, Li H, Luo M, Dong J, Chen Z, Deng X. The effects of subinhibitory concentrations of costus oil on virulence factor production in Staphylococcus aureus . J Appl Microbiol 2010; 110: 333-340
    PubMedGoogle Scholar
  • 19 Xiang H, Qiu JZ, Wang DC, Jiang YS, Xia LJ, Deng XM. Influence of magnolol on the secretion of alpha-toxin by Staphylococcus aureus . Molecules 2010; 15: 1679-1689
    CrossrefPubMedGoogle Scholar
  • 20 Cheung AL, Zhang G. Global regulation of virulence determinants in Staphylococcus auresus by SarA protein family. Front Biosci 2002; 7: 1825-1842
    CrossrefPubMedGoogle Scholar
  • 21 Novick RP, Ross HF, Projan SJ, Kornblum J, Kreiswirth B, Moghazeh S. Synthesis of staphylococcal virulence factors is controlled by a regulatory RNA molecule. EMBO J 1993; 12: 3967-3975
    PubMedGoogle Scholar
  • 22 Lorenz D, Lücker PW, Mennicke WH, Wetzelsberger N. Pharmacokinetic studies with silymarin in human serum and bile. Methods Find Exp Clin Pharmacol 1984; 6: 655-661
    PubMedGoogle Scholar
  • 23 Kielian T, Cheung A, Hickey WF. Diminished virulence of an alpha-toxin mutant of Staphylococcus aureus in experimental brain abscesses. Infect Immun 2001; 69: 6902-6911
    CrossrefPubMedGoogle Scholar
  • 24 Nilsson IM, Hartford OM, Foster T, Tarkowski A. Alpha-toxin and gamma-toxin jointly promote Staphylococcus aureus virulence in murine septic arthritis. Infect Immun 1999; 67: 1045-1049
    PubMedGoogle Scholar
  • 25 Bramley AJ, Patel AH, OʼReilly M, Foster R, Foster TJ. Roles of alpha-toxin and beta-toxin in virulence of Staphylococcus aureus for the mouse mammary gland. Infect Immun 1989; 57: 2489-2494
    PubMedGoogle Scholar
  • 26 Wardenburg JB, Patel RJ, Schneewind O. Surface proteins and exotoxins are required for the pathogenesis of Staphylococcus aureus pneumonia. Infect Immun 2007; 75: 1040-1044
    CrossrefPubMedGoogle Scholar
  • 27 Wardenburg JB, Schneewind O. Vaccine protection against Staphylococcus aureus pneumonia. J Exp Med 2008; 205: 287-294
    CrossrefPubMedGoogle Scholar
  • 28 Qiu J, Luo M, Wang J, Dong J, Li H, Leng B, Zhang Q, Dai X, Zhang Y, Niu X, Deng X. Isoalantolactone protects against Staphylococcus aureus pneumonia. FEMS Microbiol Lett 2008; 324: 147-155
    PubMedGoogle Scholar
  • 29 Li HE, Qiu JZ, Yang ZQ, Dong J, Wang JF, Luo MJ, Pan J, Dai XH, Zhang Y, Song BL, Deng XM. Glycyrrhetinic acid protects mice from Staphylococcus aureus pneumonia. Fitoterapia 2012; 83: 241-248
    CrossrefPubMedGoogle Scholar
  • 30 Rasko DA, Moreira CG, Li de R, Reading NC, Ritchie JM, Waldor MK, Williams N, Taussig R, Wei S, Roth M, Hughes DT, Huntley JF, Fina MW, Falck JR, Sperandio V. Targeting QseC signaling and virulence for antibiotic development. Science 2008; 321: 1078-1080
    CrossrefPubMedGoogle Scholar
  • 31 Hung DT, Shakhnovich EA, Pierson E, Mekalanos JJ. Small-molecule inhibitor of Vibrio cholerae virulence and intestinal colonization. Science 2005; 310: 670-674
    CrossrefPubMedGoogle Scholar
  • 32 Paul M, Leibovici L. Combination antimicrobial treatment versus monotherapy: the contribution of meta-analyses. Infect Dis Clin North Am 2009; 23: 277-293
    CrossrefPubMedGoogle Scholar
  • 33 Cegelski L, Marshall GR, Eldridge GR, Hultgren SJ. The biology and future prospects of antivirulence therapies. Nat Rev Microbiol 2008; 6: 17-27
    CrossrefPubMedGoogle Scholar
  • 34 Ohlsen K, Ziebuhr W, Koller KP, Hell W, Wichelhaus TA, Hacker J. Effects of subinhibitory concentrations of antibiotics on alpha toxin (hla) gene expression of methicillin-sensitive and methicillin-resistant Staphylococcus aureus isolates. Antimicrob Agents Chemother 1998; 42: 2817-2823
    PubMedGoogle Scholar
  • 35 Worlitzsch D, Kaygin H, Steinhuber A, Dalhoff A, Botzenhart K, Döring G. Effects of amoxicillin, gentamicin, and moxifloxacin on the hemolytic activity of Staphylococcus aureus in vitro and in vivo . Antimicrob Agents Chemother 2001; 45: 196-202
    CrossrefPubMedGoogle Scholar