1. Islamic Azad University of Medical Sciences 2. Pasteur Institute of Iran, 3. Islamic Azad University of Medical Sciences
The human gastrointestinal tract (git) has a complex community of microrganisms which is known as a gut microbiota . gut microbiota are affect variuse in essential physiological mechanisms including glucose / lipid metabolism . one of the human factors which is affected by gut microbiota is the fasting induced adipose factor (fiaf). fiaf activation is regulatey pathway which is control lpl activaion
over activation of lpl resulted in increase fat storage which is involved obesity related process. bifidobacterium bifidum, belongs to the actinobacteria phylum and member of the bifidobacteriaceae family b. bifidum gram-positive bacterium that is not motile, anaerobic, rod-shaped and not spore-forming .which is play a significant role in gut microbiota host interaction, accordingly we focused on induction effect of bacterial responses on the expression of fiaf gene in the caco2 cell line intestinal epithelial cell line.
in this study , bifidobacterium bifidum was cultured in the rogosa and sharpe agar(mrs agar) at anaerobic atmosphere including 80% co2 , 10% h2 , 10% n₂ . human intestinal epitelial cells , caco (ibrc 3010094) is growen by using dmem high glucose , which is supplemented with fbs and penicillin streptomycin. after overnight treatment of caco-2 cells with b.bifidum in multiplicity of infection(moi) = 74, total rna was extracted using thiazole solution (rnx) . rna was transcribed to cdna using cdna fermentas synthesis kits. the rt- quantitative pcr (qpcr) technique based on cyber green method was performed to assesy expression of fiaf gen . gene expression were evaluated based on delta delta ct (∆∆ct) method and gapdh as house keeping gene finally, the significance of the results is determined by independent sample t-test by spss and prism.
Our rt-qpcr result showed that, the fiaf mrna level was increased by bifidobacterium bifidum in caco2 cell line at moi 74. this up regulatory effect of b.bifidum on fiaf gene expression was significant based on p<0.05 is meaningful .
Considering the importance role of bifidobacterium bifidum in host interaction especially metabolism and role of fiaf in obesity-related processes in, according to the study which is shown the ability of b.bifidum to upregulation the expression of the fiaf gene , understanding importance of the gut microbiota members can be helpful for treatment of obesity and sickness associated with it.
Bifidobacterium bifidum , fasting induced adipose factor(fiaf), caco2 cell line , gut microbiota , o