In this study we have investigated the impact of Streptococcus thermophiluds on the rat colonic epithelium. After generation of the model axenic rat and inoculated with S. thermophilus we have investigated the interplay between bacteria and host colon. Colonic epithelium gene expression was investigated also, with two different approaches: Suppressive Subtractive Hybridization. The subtraction library was prepared subtracting mRNA between epithelial cells from colonic mono-associated rats and germ-free rats. The transcripts generated by SSH were grouped into divers Functional groups: cell-cycle arrest and induction of differentiation; cell-communication and binding. Tow candidates genes were privileged, krupel like factor 4 and 14-3-3σ. These candidate genes were tested by RT-PCR, qRT-PCR and Western blot in mono-associated animals and in germ-free animals. The tests confirmed that candidate genes increase their expression in the presence of S. thermophilus. Microarray analysis. Gene expression was measured in tow groups of animals: i) germ-free rats; ii) mono-associated rats inoculated with LMD9 strain of Streptococcus thermophilus. The results of microarray analysis data show that Streptococcus thermophilus remarkably affected gene expression in the colonic epithelial cells. Streptococcus thermophilus enhanced the expression of genes involved in different pathways in the host, compared to the gem free group.
In questo studio abbiamo analizzato l'impatto dellethermophiluds Streptococcus sull'epitelio del colon di ratto. Dopo la generazione del modello di ratto axenico e inoculato con S. thermophilus abbiamo investigato l'interazione tra il batterio e epithelo del colon di ratto. Dopo questo studio integrativo abbiamo analizzato l’espressione genica del colon usando due diversi approcci: Ibridazione sottrattiva I trascritti ottenuti dopo il sequenziamento dei cloni ottenuti con la SSH sono stati raggruppati in diversi categorie funzionali: arresto del ciclo cellulare e induzione del differenziamento, comunicazione cellulare e binding . due geni candidati sono stati privilegiati, krupel like fattore 4 e 14-3-3σ. Questi geni candidati sono stati analizzati mediante RT-PCR, qRT-PCR e Western Blot in animali mono-associati e in animali germ-free. I test hanno confermato che l’espressione dei geni candidati aumenta in presenza di S. thermophilus. Microarray Analysis. L’spressione genica è stata misurata per due gruppi di animali: i) ratti privi di germi, ii) ratti mono-associati con il ceppo LMD9 di Streptococcus thermophilus. I risultati delle analisi dei dati di microarray indicano che Streptococcus thermophilus influenza notevolmente l'espressione genica nelle cellule epiteliali del colon.
Chegdani, F., Effects of Streptococcus Thermophilus Bacteria on rat gene expression profiles, 2011-02-24 [https://hdl.handle.net/10807/284987]
Effects of Streptococcus Thermophilus Bacteria on rat gene expression profiles
Chegdani, Fatima
2011
Abstract
In this study we have investigated the impact of Streptococcus thermophiluds on the rat colonic epithelium. After generation of the model axenic rat and inoculated with S. thermophilus we have investigated the interplay between bacteria and host colon. Colonic epithelium gene expression was investigated also, with two different approaches: Suppressive Subtractive Hybridization. The subtraction library was prepared subtracting mRNA between epithelial cells from colonic mono-associated rats and germ-free rats. The transcripts generated by SSH were grouped into divers Functional groups: cell-cycle arrest and induction of differentiation; cell-communication and binding. Tow candidates genes were privileged, krupel like factor 4 and 14-3-3σ. These candidate genes were tested by RT-PCR, qRT-PCR and Western blot in mono-associated animals and in germ-free animals. The tests confirmed that candidate genes increase their expression in the presence of S. thermophilus. Microarray analysis. Gene expression was measured in tow groups of animals: i) germ-free rats; ii) mono-associated rats inoculated with LMD9 strain of Streptococcus thermophilus. The results of microarray analysis data show that Streptococcus thermophilus remarkably affected gene expression in the colonic epithelial cells. Streptococcus thermophilus enhanced the expression of genes involved in different pathways in the host, compared to the gem free group.File | Dimensione | Formato | |
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