Making weight is a practice often used in combat sports. This consists of a rapid weight loss (RWL) and a subsequent rapid weight gain (RWG) in the days preceding competition. However, this practice is often carried out based on anecdotal information provided by ex-athletes or non-professionals, which has led to several adverse events. This study aimed to assess the acute effects of a supervised nutritional period of RWL/RWG on health markers, hormone concentrations, and body composition. We performed a single-arm repeated-measures (baseline, after RWL and after RWG) clinical trial with twenty-one (8F:16M) Italian Muay Thai fighters. Body mass was significantly lower after the RWL (-4.1%) while there was a significantly higher glucose availability after RWL and RWG. Blood urea nitrogen, lipid profile, and creatinine were within the normal range after RWL/RWG. Testosterone decrease significantly after RWL and RWG in the men group. Male fighters had a significant reduction in thyroid-stimulating hormone concentration after the RWL and RWG intervention, but no change was found in women at pre-competition. Bioelectrical parameters were almost fully restored after RWG. An evidence-based and individualized nutrition methodology reduces the adverse events after an RWL and RWG practice, although the impact on the hormonal profile is inevitable.