Planta Medica, Table of Contents

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^*

¹College of Quartermaster Technology, Jilin University, Changchun, PR China

,

Jing Dong^*

²Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin University, Changchun, PR China

,

Xiaohan Dai

²Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin University, Changchun, PR China

,

Yu Zhang

²Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin University, Changchun, PR China

,

Jianfeng Wang

²Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin University, Changchun, PR China

,

Hongen Li

²Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin University, Changchun, PR China

,

Chongjian Lu

²Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin University, Changchun, PR China

,

Wei Tan

²Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin University, Changchun, PR China

,

Xiaohan Gao

¹College of Quartermaster Technology, Jilin University, Changchun, PR China

,

Xuming Deng

²Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin University, Changchun, PR China

,

Shijin Bu

³College of Veterinary Medicine, Yangzhou University, Yangzhou, PR China

,

Xiaodi Niu

¹College of Quartermaster Technology, Jilin University, Changchun, PR China

› Author Affiliations

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

Key words

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

References
1 Lowy FD. Staphylococcus aureus infections. N Engl J Med 1998; 339: 520-532
2 Rubinstein E, Kollef MH, Nathwani D. Pneumonia caused by methicillin-resistant Staphylococcus aureus . Clin Infect Dis 2008; 46: 378-385
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
4 Rasko DA, Sperandio V. Anti-virulence strategies to combat bacteria-mediated disease. Nat Rev Drug Discov 2010; 9: 117-128
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
6 Kobayashi SD, DeLeo FR. An update on community-associated MRSA virulence. Curr Opin Pharmacol 2009; 9: 545-551
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
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
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
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
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
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
13 Wellington K, Jarvis B. Silymarin: a review of its clinical properties in the management of hepatic disorders. BioDrugs 2001; 15: 465-489
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
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
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
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
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
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
20 Cheung AL, Zhang G. Global regulation of virulence determinants in Staphylococcus auresus by SarA protein family. Front Biosci 2002; 7: 1825-1842
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
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
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
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
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
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
27 Wardenburg JB, Schneewind O. Vaccine protection against Staphylococcus aureus pneumonia. J Exp Med 2008; 205: 287-294
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
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
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
31 Hung DT, Shakhnovich EA, Pierson E, Mekalanos JJ. Small-molecule inhibitor of Vibrio cholerae virulence and intestinal colonization. Science 2005; 310: 670-674
32 Paul M, Leibovici L. Combination antimicrobial treatment versus monotherapy: the contribution of meta-analyses. Infect Dis Clin North Am 2009; 23: 277-293
33 Cegelski L, Marshall GR, Eldridge GR, Hultgren SJ. The biology and future prospects of antivirulence therapies. Nat Rev Microbiol 2008; 6: 17-27
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
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