Experimental research on leakage rate of silastic O-ring at simulative space environment is conducted to investigate the effect of compression ratio, temperature, load decay of O-ring and radialization of atomic oxygen on the leakage rate of O-ring. The results show that the leakage rate of O-ring decreases with the increase of compression ratio, increases with the rise of temperature, and it is affected by the load decay of O-ring and radialization of atomic oxygen at various degrees.