Chronic hepatitis C (CHC) affects the activity of NK cells, but successful interferon-free treatment partially restores it. The goal of the study was to assess whether gender influences those alterations. We examined 21 women after menopause and 24 men with CHC treated with directly acting antivirals (DAA), and 33 healthy volunteers. With flow cytometry, we analysed KIR2DS4, NKG2D, NKp30, KIR2DL2/DL3, NKG2A, TRAIL and granzyme B on the surface of NK cells, simultaneously we checked serum CXCL10 with ELISA. Overall, patients with CHC had higher expression of KIR2DS4, NKG2A, NKp30 and a lower percentage of NK cells among lymphocytes than the control group. After the treatment KIR2DS4, KIR2DL2/DL3 and NKG2A, TRAIL NKp30 on NK cells decreased, while the percentage of NK cells, expression of granzyme B and NKG2D increased. Serum CXCL10 was elevated before the treatment and dropped afterwards. We observed differences between genders in the expression of KIR2DL2/DL3 (higher in female) and NKp30 (elevated in men) comparing CHC/control groups. After the treatment of KIR2DL2/DL3, NKp30 and CXCL10 dropped only in female while granzyme B increased in male. In conclusion, the response of NK cells among men and women in post-menopausal age with CHC differs. Our research may lead to more studies on different nature of female and male immune systems in the context of HCV infection and treatment.