Aim: Pentoxifylline (PTX) is a methylxanthine derivative medicine used to improve motility of human spermatozoa in-vitro. It is commonly used in treatment of male-factor infertility, including asthenozoospermia. This study aimed to evaluate the effect of PTX on human sperm parameters and DNA integrity from asthenozoospermic problem.Material and methods: A total of 38 infertile men with asthenozoospermia were allocated in this experimental study. Specimens were randomly divided into experimental group treated with 3.6 mM PTX, and control group. All samples were incubated at 37˚ C for 45 min. Semen parameters and sperm DNA fragmentation were measured using sperm chromatin dispersion (SCD) test.Results: PTX improved sperm motility, significantly, compared to the control (85.76±5.96 Vs 79.44±9.37, respectively, P<0.01). There was also a significant decrease in sperm viability in the PTXtreated group in comparison to controls (87.7±8.3 Vs 83.5±9, respectively, P<0.01). In addition, sperm DNA fragmentation was higher in PTX-treated group compared to control (23.36±10.25 and 18.5±8.74, respectively, P<0.0001).Conclusion: PTX might have some negative impact (s) on sperm DNA quality, although it has improved the sperm motility. Further studies are needed to elucidate the safety of PTX treatments in ART clinics.