Assessment Of Toll Like Receptor 4 Activation in HEK Transformed Cell Line via Incubation with Semen from Asthenozoospermia Patients

Background: Infertility could disrupt the essential steps of hu-man life as an important disorder in the evolution pathway and results in an extensive trauma. Asthenozoospermia is one of the prevalent causes of male infertility which could be a result of infection. Infection reduces the sperm motility and causes the male infertility. In the studies carried out on semen and sperm, it is clarified that bacterial endotoxins such as the LPS of gram-negative bacteria, activates the TLR4 pathway and induces the expression of Chemokines and Cytokines coding genes. Con-sequently, it has an important role in apoptosis induction and sperm motility reduction. Sperm motility reduction often oc-curs as a result of infection and TLR4 induction. According to this fact, the goal of this study is to benefit from a suitable cellu-lar model such as transfected human embryonic kidney (HEK) cells with TLR4, in order to study the effects of asthenozoo-spermia patients semen and to prove the existence of specified infection for this receptor from assessment of gene expression TLR4 downstream of iNOS and TNF-α in asthenozoospermia patients.Materials and Methods: Semen samples of 15 in asthenozoo-spermia patients and 15 healthy person who refer to the Isfahan fertility and infertility clinic, were examined. Motility of sam-ples was studied according to the WHO 2010 protocol. LPS was studied as the positive control. Transfected HEK cells with TLR4 were treated with different concentrations of semen. After 24 hours, in order to assess the afformentioned genes, mRNAs were extracted by means of TRIZOL, cDNAs were synthesized and then Real time PCR was carried out. Results: Significant increment in expression of downstream genes of TLR4 receptor including iNOS and TNF-α in asthenozoospermia patients in comparison to healthy people was found out.Results: Significant increment in expression of downstream genes of TLR4 receptor including iNOS and TNF-α in astheno-zoospermia patients in comparison to healthy people was found out.Conclusion: In conclusion, infection induces the activation of the TLR4 pathway, reduces the sperm motility and results in infertility

Keywords:

Asthenozoospermia , Transfected HEK Cells , TLR4 Pathway , Infection , LPS