In order to explore the effects of combined stress of temperature and vibration intensity on tissue damage and biochemical indexes of sea bass (Lateolabrax japonicus) during in-water keep-alive transportation, six experimental groups were designed with transportation temperature of 20 °C and 12 °C and vibration intensity of 0, 20 and 50 r/min as variables. After simulated transportation for 48 h, the effects of temperature and vibration stress on L. japonicus during simulated transportation were studied by analyzing oxidative stress, tissue damage and immune indexes. The results show that: ① The vibration intensity of different temperature ratios has great influence on the survival rate of L. japonicus. After 24 h transportation, the survival rates of the transportation group at 20 °C at 0, 20 and 50 r/min were 72.70, 62.50 and 18.2% respectively. However, the survival rate of all transport groups at 12 °C is 100%. ② The change trend of each index is different at different temperatures. Under the transport conditions of 20 °C, catalase (CAT), glutathione-S-transferase (GST), malondialdehyde (MDA), glutamic acid transaminase (ALT), aspartate transaminase (AST), acid phosphatase (ACP), alkaline phosphatase (AKP), and lysozyme (LZM) increased with the increase of transport time, and superoxide dismutase (SOD), immunoglobulin ( IgM) showed a tendency to increase and then decrease with increasing transport time, and blood glucose (GLU) showed a tendency to decrease with increasing transport time. Among them, low temperature treatment can reduce the oxidative stress, tissue damage and immune system damage of L. japonicus. At the transportation temperature of 12 °C, the activities of CAT, MDA, ALT, AST and LZM increased first and then decreased, while the activities of SOD, GST, GLU and IgM decreased first and then increased. The highest value of SOD was 0.64 U/mgprot in the transportation groups at 20 °C and 50 r/min, but only 0.37 U/mgprot in the transportation group at 12 °C and 50 r/min. The research shows that the L. japonicus should be transported at low temperature, the temperature should be kept at 12 °C, and the flat transportation road should be chosen to reduce vibration frequency during transportation. This study explored the effects of combined stress of temperature and vibration intensity on marine fish. The results can provide reference for short-distance transportation of L. japonicus and improve the survival rate of live fish during transportation.