Background: Nicotine as a free radical reduces male fertility. Green tea, as an antioxidant, has contributed to protecting testes function; however, green tea kombucha has never been studied for such benefits. This study demonstrated the effect of green tea kombucha in increasing Leydig cells number and testosterone levels in male Wistar rats exposed to nicotine from electric cigarettes. Methods: A post-test-only control group trial was performed on 28 male Wistar rats randomly divided into control, untreated, treatment groups 1 and 2. The control group was exposed to e-cigarette vapor + 1 ml of distilled water. Treatment groups 1 and 2 were exposed to e-cigarette vapor + 0.5 ml/day and 1 ml/day of green tea kombucha, respectively—the e-cigarette vapor exposure occurred daily for 30 minutes. After 28 days, Leydig cell and testosterone levels were measured and compared between groups. Results: The control group's mean value of Leydig cell number is lower than untreated group (p=0.004). Only treatment group 2 had a higher mean of Leydig cell number than control (p<0.001). The control group's mean value for testosterone levels was lower than untreated group (p<0.001). The mean testosterone level in treatment groups 1 and 2 was higher than in untreated and control (p<0.001). In addition, treatment group 2 had higher mean testosterone levels than group 1 (p<0.001). Conclusions: Green tea kombucha has been shown to inhibit the harmful effects of nicotine. These results serve as a basis for the administration of green tea kombucha to prevent free radical-induced testosterone decline. Keywords: Kombucha, Leydig Cells, Testosterone, Nicotine, Antioxidant