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| Status |
Public on Nov 22, 2011 |
| Title |
The Apple Flavonoid Phloretin Inhibits Escherichia coli O157:H7 Biofilm Formation and Ameliorates Colon Inflammation in Rat |
| Organism |
Escherichia coli |
| Experiment type |
Expression profiling by array
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| Summary |
Pathogenic biofilms have been associated with persistent infections due to high resistance to antimicrobial agents while commensal biofilms often fortify host immune system. Hence, controlling biofilm formation of both pathogenic bacteria and commensal bacteria is important in bacteria-related diseases. We investigated the effect of plant flavonoids on biofilm formation of both enterohemorrhagic Escherichia coli O157:H7 and three commensal E. coli K-12 strains. Phloretin abundant in apples markedly reduced E. coli O157:H7 biofilm formation without affecting the growth of planktonic cells while phloretin did not harm commensal E. coli K-12 biofilms. Also, phloretin reduced E. coli O157:H7 attachment to human colon epithelial cells. Global transcriptome analyses revealed that phloretin repressed toxin genes (hlyE and stx2), autoinducer-2 importer genes (lsrACDBF), a curli gene (csgA), and a dozens of prophage genes in E. coli O157:H7 cells. Electron microscopy confirmed that phroretin reduced the curli production in E. coli O157:H7. In addition, phloretin suppressed TNF-α-induced inflammatory response in vitro using human colonic epithelial cells. Moreover, in the trinitrobenzene sulfonic acid (TNBS)-induced rat colitis model, phloretin significantly ameliorated colon inflammation and body weight loss. Taken together, our results suggest that phloretin may act as an inhibitor of E. coli O157:H7 biofilm formation as well as anti-inflammatory agent on inflammatory bowel diseases while leaving beneficial commensal E. coli biofilm intact.
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| Overall design |
For the microarray experiments, E. coli O157:H7 EDL933 was inoculated in 25 ml of LB in 250 ml flasks with overnight cultures that were diluted at 1:100. Cells were shaken at 100 rpm and 37°C for 7 hrs. Cells were immediately chilled with dry ice and 95% ethanol (to prevent RNA degradation) for 30 sec before centrifugation in 50 ml centrifuge tubes at 13,000 g for 2 min; cell pellets were frozen immediately with dry ice and stored -80°C. RNA was isolated using Qiagen RNeasy mini Kit (Valencia, CA, USA). To eliminate DNA contamination, Qiagen RNase-free DNase I was used to digest DNA. RNA quality was assessed by Agilent 2100 bioanalyser using the RNA 6000 Nano Chip (Agilent Technologies, Amstelveen, The Netherlands), and quantity was determined by ND-1000 Spectrophotometer (NanoDrop Technologies, Inc., DE, USA).
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| Contributor(s) |
Lee J, Lee J, Kim J, Regmi SC, Cho MH, Yun H, Lee C |
| Citation(s) |
21930760 |
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| Submission date |
Mar 24, 2011 |
| Last update date |
Mar 08, 2019 |
| Contact name |
Jintae Lee |
| E-mail(s) |
jtlee@ynu.ac.kr
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| Phone |
82-53-810-2533
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| Organization name |
Yeungnam University
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| Department |
Chemical engineering
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| Lab |
Biotechnology
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| Street address |
214-1 Daedong
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| City |
Gyeongsan-Si |
| State/province |
Gyeongsangbuk-Do |
| ZIP/Postal code |
712-749 |
| Country |
South Korea |
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| Platforms (1) |
| GPL3154 |
[E_coli_2] Affymetrix E. coli Genome 2.0 Array |
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| Samples (2) |
| GSM697301 |
E. coli O157:H7 EDL933 in LB at 37oC at 7 hrs incubation |
| GSM697302 |
E. coli O157:H7 EDL933 in LB at 37oC at 7 hrs incubation with phloretin |
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| Relations |
| BioProject |
PRJNA139557 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE28144_RAW.tar |
1.9 Mb |
(http)(custom) |
TAR (of CEL, CHP) |
| Processed data included within Sample table |
| Processed data provided as supplementary file |
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