The Immunomodulatory Role of G2013 (α-L-Guluronic Acid) on the Expression of TLR2 and TLR4 in HT29 cell line.
BACKGROUND
The non-steroidal anti-inflammatory drugs (NSAIDs) play crucial role in the controlling of inflammatory diseases. Due to the vast side effects of NSAIDs, its use is limited. G2013 or α-L-Guluronic Acid is a new NSAID with immunomodulatory features.


OBJECTIVES
Considering the leading role of TLRs in inflammatory responses, in this study, we aimed to evaluate G2013 cytotoxicity and its effect on the expression of TLR2 and TLR4 molecules.


METHODS
HEK293-TLR2 and HEK293-TLR4 cells were cultured and seeded on 96-well cell plate, and MTT assay was performed for detecting the viability of the cells after treatment with different concentrations of G2013. HT29 cells were grown and treated with low and high doses of G2013. After total RNA extraction and cDNA synthesis, quantitative real-time PCR were performed to assess the TLR2 and TLR4 mRNA synthesis.


RESULTS
We found that concentrations of ≤125 µg/ml of G2013 had no apparent cytotoxicity effect on the HEK293-TLR2 and -TLR4 cells. Our results indicated that after G2013 treatment (5 µg/ml) in HT29 cells, TLR2 and TLR4 mRNA expression decreased significantly compared with the untreated control group (p=0.02 and p=0.001 respectively).


CONCLUSION
The results of this study revealed that G2013 can down regulate the TLR2 and TLR4 gene expression and exerts its inhibitory effect. Our findings are parallel to our previous finding which showed G2013 ability to down regulate the signaling pathway of TLRs. However, further studies are needed to identify the molecular mechanism of G2013.
['Farhang|Hamid|H|', 'Sharifi|Laleh|L|', 'Dallal|Mohammad Mehdi Soltan|MMS|', 'Moshiri|Mona|M|', 'Norouzbabaie|Zahra|Z|', 'Bokaie|Saied|S|', 'Aletaha|Somaye|S|', 'Zargar|Seyed Jalal|SJ|', 'Mirshafiey|Abbas|A|']
D002470:Cell Survival D005786:Gene Expression Regulation D057809:HEK293 Cells D019073:HT29 Cells D006603:Hexuronic Acids D006801:Humans D007155:Immunologic Factors D012333:RNA, Messenger D051195:Toll-Like Receptor 2 D051197:Toll-Like Receptor 4
G2013 TLR2 TLR4 HT29 NSAIDs α-L-Guluronic Acid.