Plastic covers have been used to prevent environmental constraints negatively affecting sweet cherry production in Southern Chile. However, less information is available on agronomic practices and their effects on fruit quality in sweet cherry covered orchards. Thus, in this study, we evaluated the impact of fruit load regulation on cherries’ antioxidant-related parameters and the quality and condition at harvest and postharvest in sweet cherry (Prunus avium) cv. Regina that was cultivated under a plastic cover in Southern Chile. For this, four fruit load treatments were manually applied—(i) 100% fruit load (the control), (ii) 80% fruit load, (iii) 60% fruit load, and (iv) 40% fruit load—in a commercial sweet cherry orchard for two seasons (2021/2022 and 2022/2023). The results revealed that the yield and fruit load were not significantly different between the treatments. Interestingly, the 60% and 40% fruit loads increased the fresh weight, fruit size, and firmness (20.3%) compared to the control (the 100% fruit load) during both seasons. Likewise, the 60% and 40% fruit load treatments exhibited the highest fruit size distribution of 30 mm, while the 100 and 80% fruit load treatments showed the highest fruit distribution with fruit sizes between 28 mm and 24 mm. The total soluble solids (TSSs) did not vary among the fruit load treatments, while a significant increase was found in the titratable acidity (TA) in the 60 and 40% fruit load treatments during both seasons. No significant differences in antioxidant activity (AA) and total phenols (TPHs) among the treatments were observed during both seasons. Overall, the results revealed that the fruit load treatments, mainly 40%, increased the fruit weight and firmness and reduced pitting in fruits by 39.4% at postharvest. Thus, fruit thinning might be an important agronomical practice to regulate fruit load, positively affecting fruit quality at harvest and during postharvest storage in sweet cherry cv. Regina cultivated under a plastic cover. However, more biochemical and molecular studies are needed to elucidate the mechanism involved in this improvement.