In order to prepare andrographolide ethosomes, screen the optimal prescription, and evaluate its quality, this study used the injection method to prepare andrographolide ethosomes. With encapsulation efficiency as the evaluation index, the formulation was optimized using the Box-Behnken response surface methodology, and the morphology, particle size distribution, and zeta potential were characterized. The preliminary stability was assessed using the leakage rate as an indicator. The in vitro transdermal properties of andrographolide ethosomes were investigated using Franz diffusion cells, and the effects of different drug administration forms on the transdermal penetration of andrographolide were compared. The results showed that the optimal formulation for preparing andrographolide ethosomes consisted of 1.40% soybean lecithin, 25.45% ethanol, and 0.14% drug content. The prepared andrographolide ethosomes were round in appearance, with an average encapsulation efficiency of 70.08%±0.57%, an average particle size of 191.81±4.57 nm, a polydispersity index of 0.130±0.020, and a zeta potential of -45.8±0.51 mV. The ethosomes exhibited good stability when stored at 4 °C for three months, with a leakage rate of 6.95%±0.63%. The steady-state transdermal permeation rate of andrographolide ethosomes was 1.33±0.25 μg/(cm²·h), which was 2.11 times higher than that of andrographolide liposomes and 2.66 times higher than that of andrographolide ethanol aqueous solution. The drug skin retention of andrographolide ethosomes was 14.92±1.09 μg/cm², which was 4.24 times higher than that of andrographolide liposomes and 16.74 times higher than that of andrographolide ethanol aqueous solution.The results indicate that the preparation process of andrographolide ethosomes is simple, quality controllable, and exhibits good stability and transdermal absorption performance, showing promising application prospects.