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26 février 2011 6 26 /02 /février /2011 10:28

Toxicon. 2010 Dec 22. [Epub ahead of print]

Allicin from garlic neutralizes the hemolytic activity of intra- and extra-cellular pneumolysin O in vitro.

Arzanlou MBohlooli SJannati EMirzanejad-Asl H.

Department of Microbiology, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran.


Pneumolysin (PLY) is a key virulence factor contributes to the pathogenesis of Streptococcus pneumoniae. In this study we investigated the effect of allicin and aqueous garlic extracts on hemolytic activity of PLY both in prelysed and intact cells. Additionally the antimicrobial activity of allicin was tested against the bacteria. All tested materials potently inhibited the PLY hemolytic activity. Allicin neutralizes PLY in a concentration- and time-dependent manner. Twenty five minute incubation of PLY (2 HU/mL) with 0.61 μM/mL concentration of allicin, totally inhibited hemolytic activity of PLY (IC50 = 0.28 μM/mL). The inhibitory activity of old extract of garlic was similar to pure allicin (IC50 = 50.46 μL/mL; 0.31 μM/mL; P < 0.05). In contrast fresh extract of garlic inhibits the PLY hemolytic activity at lower concentrations (IC50 = 13.96 μL/mL; 0.08 μM/mL allicin). Exposure of intact cells to allicin (1.8 μM) completely inhibited hemolytic activity of PLY inside bacterial cells. The inhibitory effect of the allicin was restored by addition of reducing agent DTT at 5 mM, proposing that allicin likely inhibits the PLY by binding to cysteinyl residue in the binding site. The MIC value of allicin was determined to be 512 μg/mL (3.15 μM/mL). These results indicate that PLY is a novel target for allicin and may provide a new line of investigation on pneumococcal diseases in the future.

Copyright © 2010 Elsevier Ltd. All rights reserved.

PMID: 21184771 [PubMed - as supplied by publisher]

 

 

 

J Med Microbiol. 2010 Sep;59(Pt 9):1044-9. Epub 2010 Jun 10.

Inhibition of streptolysin O by allicin - an active component of garlic.

Arzanlou MBohlooli S.

Department of Microbiology, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran. m.arzanlou@arums.ac.ir

Streptolysin O (SLO) is a potent cytolytic toxin produced by almost all strains of group A streptococci and is considered an important virulence factor for this organism. In this study we investigated the effect of allicin and aqueous garlic extracts on the haemolytic activity of SLO. All tested materials potentially inhibited the SLO haemolytic activity. Allicin neutralized SLO in a dose- and time-dependent manner. A 15 min incubation of SLO with 35 microg allicin totally inhibited the haemolytic activity of SLO [IC(50) (concentration necessary to reach half maximum inhibition)=5.97 microg]. The inhibitory activity of an old extract of garlic was equipotent to pure allicin (IC(50)=6.27 microg; P<0.05). In contrast, fresh extract of garlic inhibited the SLO haemolytic activity at lower concentrations (IC(50)=1.59 microl; 1.9 microg allicin). The inhibitory effect of the allicin was restored by addition of reducing agent DTT at 2 mM, suggesting that allicin likely inhibits the SLO by binding to the cysteine residue in the binding site. These results indicate a new activity for allicin and allicin may be a potential alternative drug against streptococcal diseases.

PMID: 20538890 [PubMed - indexed for MEDLINE]


 Thromb Res. 2009 Sep;124(4):477-82. Epub 2009 Jul 26.

Allicin and disulfiram enhance platelet integrin alphaIIbbeta3-fibrinogen binding.

Manaster YShenkman BRosenberg NSavion N.

Goldschleger Eye Research Institute and Department of Human Molecular Genetics and Biochemistry, Sackler Faculty of Medicine, Tel Aviv University, Israel.

INTRODUCTION: Activation of the platelet receptor alphaIIbbeta3 (glycoprotein IIbIIIa) involves a change in the disulfide bonds pattern in the extra-cellular domain of the receptor. The disulfide-bond reducing agent, dithiothreitol (DTT), can increase integrin activity, and point mutations of specific cysteine residues of the integrin can cause its lockage at the high affinity state. The present study is aimed to support the hypothesis that prevention of specific alphaIIbbeta3 intra-molecular disulfide bond formation increases receptor-ligand binding activity.

METHODS: Platelet aggregation was induced by collagen or ADP and epinephrine. Integrin alphaIIbbeta3-fibrinogen binding was evaluated on prostaglandins E(1) (PGE(1))-treated washed platelets or baby hamster kidney (BHK) cells expressing human alphaIIbbeta3. Integrin was directly activated by an anti-ligand induced binding site (LIBS) PT25-2 antibody. The effect of sulfhydryl-reactive agents, such as allicin, glutathione, dithiobis nitrobenzoic acid (DTNB) and disulfiram, was tested on alphaIIbbeta3 activity.

RESULTS: Allicin (40 microM) completely inhibited washed platelets agonist-induced aggregation. Both allicin and disulfiram (40 microM) inhibited alphaIIbbeta3-fibrinogen binding and P-selectin expression in washed platelets. However, there was an increase in alphaIIbbeta3-fibrinogen binding but not P-selectin expression in PGE(1)-treated washed platelets activated by PT25-2 antibody. At a high concentration (400 microM) both inhibited alphaIIbbeta3-fibrinogen binding. Similarly, in BHK cells expressing alphaIIbbeta3 activated by PT25-2 antibody, allicin at a low concentration increased alphaIIbbeta3 activity.

CONCLUSIONS: Allicin and disulfiram inhibit agonist-induced washed platelet activation probably via inhibition of platelet signaling, but enhance PT25-2 antibody-induced alphaIIbbeta3 integrin activity most likely by preventing reformation of disulfide bridges thereby stabilizing the active conformation of the integrin.

PMID: 19632706 [PubMed - indexed for MEDLINE]

 

J Antimicrob Chemother. 2009 Jan;63(1):151-4. Epub 2008 Nov 11.

In vitro activity of an aqueous allicin extract and a novel allicin topical gel formulation against Lancefield group B streptococci.


Cutler RROdent MHajj-Ahmad HMaharjan SBennett NJJosling PDBall VHatton PDall'Antonia M.

University of East London, Romford Rd, Stratford, London E15 4LZ, UK. rcutler@uel.ac.uk

BACKGROUND: Studies have shown the efficacy of intra-partum antibiotics in preventing early-onset group B streptococcal sepsis. This approach results in a high intra-partum antibiotic use. Worryingly, the same antibiotics used in prophylaxis are also first-line treatment for neonatal sepsis, and antibiotic exposure in the peri-natal period has been shown to be a risk factor for late-onset serious bacterial infections and allergic disease. Antibiotic exposure in the peri-natal period is becoming a major public health issue; alternative strategies are needed. Garlic has been traditionally used to treat vaginal infections. Allicin is the main antibacterial agent isolated from garlic.

OBJECTIVES: The aim of the study was to investigate the in vitro activity of a novel allicin extract in aqueous and gel formulation against 76 clinical isolates of Lancefield group B streptococci (GBS).

METHODS: MICs and MBCs of allicin were determined for 76 GBS isolates by agar dilution and microtitre plate methods. Killing kinetics were determined for a selected 16 of the 76 strains. Agar diffusion tests were compared for allicin liquid and gel (500 mg/L).

RESULTS AND CONCLUSIONS: MICs and MBCs of allicin liquid were 35 to 95 mg/L and 75 to 315 mg/L, respectively. Time/dose kill curves produced a 2-3 log reduction in cfu/mL within 3 h and no detectable growth at 8 and 24 h. A novel 500 mg/L allicin gel produced an average zone size of 23+/-6 mm compared with 21+/-6 mm for allicin in water.

 

Aqueous allicin is bactericidal against GBS isolates and maintains activity in a novel gel formulation.


PMID: 19001449 [PubMed - indexed for MEDLINE]

 


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