One of the primary causes of male infertility is the asthenozoospermia (AZS) which is characterized by defective sperm motility. More genes have been recently associated with AS. Oxidative stress is also another notable cause for AZS. This study aimed to investigate the biochemical (oxidant/antioxidant status) and molecular (AZS-associated genes) changes in asthenozoospermic patients. To achieve this goal, we utilized spectrophotometry and real time PCR (qPCR) assays on semen samples collected from AZS patients (n = 50) and normozoospermia men (n = 25). Semen samples were categorized into these two categories based on the sperm physical characters; motility, viability, and morphological parameters. The examined parameters included the oxidative peroxide malondialdehyde (MDA), the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), and asthenozoospermic genes (CATSPER1, SEPT12, CFAP43, and CFAP44). Semen of AZS patients exhibited significantly (P <0.05) higher MDA levels, significantly (P <0.05) reduced expression of CATSPER1, SEPT12, CFAP43, and CFAP44 and activities of CAT, SOD, and GPx compared to semen of the control (normozoospermia) group. Among the four genes, only CATSPER1 and SEPT12 expression showed a significantly (P <0.05) negative correlation with abnormal sperm forms and seminal MDA levels and a significantly positive correlation with sperm motility and count and seminal levels of the antioxidant enzymes. These findings provided empirical proof that decreased CATSPER1 and SEPT12 expression and increased seminal oxidative stress are associated with asthenozoospermia.