Objective Our study is aim to investigate the possible relationship of seminal elastase, on semen parameters and DNA fragmentation in male patients of recurrent pregnancy loss (RPL) between June 2019 and January 2020. Methods The patients included 80 male patients of RPL couple and 25 male patients from couples with clinical pregnancy through in vitro fertilization due to the female tubal factor. The semen samples were used to determine the seminal elastase, computed assisted semen analysis, sperm dispersion test and sperm morphology analysis. Results Compared to the control group, the levels of seminal elastase was increased in the RPL group. The RPL group was divided into the normal group (Elastase<600 ng/mL) and abnormal group (Elastase ≤ 600 ng/mL).The abnormal group exhibited the lower percentage of progressive sperm (P=0.002) and normal morphology (P=0.009),but higher precentage of DNA fragmentation (P=0.002). Meanwhile, the seminal elastase was positively associated with DNA fragmentation (r=0.36,P=0.001), but was inversely associated with the sperm motility (r=-0.43,P<0.001) and normal morphology (r=-0.39,P<0.001). Conclusion Our study may unveil the possible effects of the seminal elastase on the semen parameters and DNA fragmentation in the male patients of RPL couples. Further studies should put more emphasis on the silent genital tract inflammation of the patients.