The development of environmentally friendly corrosion inhibitors has become a research hotspot. Aiming at the potential corrosion inhibition effect of perilla seed extract on Q235 carbon steel, the corrosion inhibition effect was quantitatively evaluated by various research methods, and the effective corrosion inhibition composition and mechanism were discussed. The research methods include potentiodynamic polarization curve method, HPLC-MS, FT-IR, XPS and chemical calculation. The experimental results show that the inhibitor prepared from perilla seed extract is a mixed inhibitor, and its adsorption behavior accords with Langmuir adsorption theory and its adsorption free energy is −22.70 kJ/mol. Combined with the experimental results and theoretical calculation, the effective corrosion inhibiting components are luteolin and apigenin. Theoretical calculation shows that both of them are adsorbed parallel to the surface of carbon steel to form thin films. The adsorption mechanism is that carbonyl O atoms in luteolin and apigenin hybridize with the 3 d empty orbit of Fe. From the point of view of quantum chemistry, the smaller the HOMO value and the energy gap value, the better the adsorption of corrosion inhibitor on the surface of carbon steel. From the point of view of molecular dynamics simulation, the greater the absolute value of adsorption energy, the better the adsorption of corrosion inhibitor on carbon steel surface.