Evaluation of TLR5 Gene Expression in Human Vaginal Epithelial Cells Infected by Trichomonas Vaginalis
Abstract
Trichomoniasis is a sexual trasmited disease caused by Trichomonas vaginalis.TLRs are a family of pattern recognition receptors expressed in various mammalian cells and have important role in pathogen detection and inflammatory responses. In present study the role of TLR5 in HeLa cells and human vaginal epithelial cells exposed to T. vaginalis was investigated.
Materials and Methods: T. vaginalis and the HeLa and human vaginal epithelial cells (HVECs) were cultured in RPMI and trypticase yeast extract maltose (TYM), respectively. After adaption of parasite and epithelial cells by RPMI-TYM medium co-culture (9:1 vol/vol), HeLa cells and HVECs were stimulated with different ratios of T. vaginalis trophozoites for 24-hour. Following RNA extraction and cDNA synthesis, the gene expression levels of TLR5, IRAK1 and NF-κB were assessed using real-time PCR. Besides, the proteins levels were measured using western blotting.
Results: Real-time PCR results showed gene expression of TLR5, IRAK1, and NF-κB in T. vaginalis exposed HVECs and HeLa cells increased significatly(P<0.05). Additionally, western blot analysis showed a statistically significant increase in TLR5, and NF-κB proteins in 1X, 3X and 5X of HeLa cells and only in 5X of HVECs exposure to the parasite (P<0.05).