In this study, it was demonstrated the ameliorative effect of zingerone (ZO) (25 and 50 mg/kg body weight) against vancomycin (VCM) (200 mg/kg body weight) administered to rats on some metabolic enzymes' activities in the lung, liver, kidney, and testis tissues of rats. Forty-two rats were divided into six groups as follows: control, ZO-25, ZO-50, VCM, VCM + ZO-25, and VCM + ZO-50. alpha-Glycosidase, butyrylcholinesterase, aldose reductase, acetylcholinesterase, paraoxonase-1, and carbonic anhydrase enzyme activities were significantly (P < .05) decreased in VCM group when compared with the control group. ZO, supplied with VCM, significantly activated some of these enzyme in all tissues. The results of this study showed that ZO regulates abnormal increases and decreases in VCM-induced metabolic enzyme activities in all tissues.