To establish a RP-HPLC method for the determination of ophiopogonin A content in Radix ophiopogonison; and to observe the intervention effect of Radix ophiopogonison extract on acute lung injury after intestinal ischemia reperfusion. Synergi 4 μ Hydro-RP 80A column (250 mm × 4.6 mm, 5 μm) is used, column temperature is 25°C, mobile phase is water-acetonitrile (volume ratio 62:38), flow rate is 1.0 mL/min and detection wavelength 208 nm. Rat model of intestinal ischemia reperfusion is established, and then the rats are randomized into control group, ischemia reperfusion group and Radix ophiopogonison intervention group. Total protein content in Bronchoalveolar Lavage Fluid (BALF), dry/wet weight ratio (D/W) of lung tissues, peroxidase (MPO) content in lung tissues and Malondialdehyde (MDA) content in plasma are measured. Mass concentration of ophiopogonin A shows a good linear relationship (r=0.9999) with the peak area at a range of 0.020~0.200 mg/mL, and the average recovery is 99.75% (RSD=0.22%, n=9). Compared with the ischemic reperfusion group, BALF total protein content increases significantly, while lung D/W decreases significantly in the intervention group. The content determination method established herein is simple and accurate, which can be used for the quality control of Radix ophiopogonison medicinal material. Radix ophiopogonison extract can significantly reduce the degree of acute lung injury.