High temperature and high relative humidity (RH) are one of the most serious agricultural meteorological disasters that limit the production capacity of agricultural facilities. However, little information is available on the precise interaction between these factors on tomato growth. The objectives of this study were to determine the effect of high temperature under different RH levels on tomato growth and endogenous hormones and to determine the optimal RH for tomato seedling growth under high temperature environment. Two high temperature (38/18 °C, 41/18 °C) and three relative humidity (50 ± 5%, 70 ± 5%, 90 ± 5%) orthogonal experiments were conducted, with 28/18 °C, 50 ± 5% (CK) as control. The results showed that the dry matter accumulation of tomato plants under high temperature environment was significantly lower than that of CK. At 38 °C, the dry matter accumulation with 70% relative humidity was not significantly different from that of CK; at 41 °C, dry matter accumulation with 70% and 90% relative air humidity was significantly greater than that of 50%. The concentrations of soluble sugar and free amino acids in all organs in high temperature-treated plants were significantly higher than that in CK. As relative humidity increased, soluble sugar concentrations of each organ decreased, and the free amino acid concentrations increased. Cytokinin (ZT) and indole acetic acid (IAA) concentrations in tomato buds were significantly lower than in CK under high temperature conditions. The lower the RH, the lower the content of ZT and IAA. The gibberellin (GA3) and abscisic acid (ABA) concentrations were higher than in CK under high temperatures. GA3 concentrations decreased and ABA concentrations were augmented with increased humidity. The differences of tomato seedling growth indices and apical bud endogenous hormone concentrations between RHs under high temperature conditions were significant. Raising RH to 70% or higher under high temperature conditions could be beneficial to the growth of tomato plants. The results contribute to a better understanding of the interactions between microclimate parameters inside a Venlo-type glass greenhouse environment, in a specific climate condition, and their effects on the growth of tomato.