Abstract:[Background] Short-chain fatty acids (SCFAs) have a wide range of physiological activities and biological effects, such as providing energy, regulating nutrient metabolism, inhibiting endogenous cholesterol synthesis, and so on. [Objective] The model of Caco-2 cell absorption SCFAs was established, which was used to research the effect of Lactobacillus on intestinal uptake of SCFAs. [Methods] The integrity and stability of Caco-2 cell model was evaluated by transepithelial electrical resistance (TEER), cell ultrastructure, apparent permeability coefficients of phenol red (Papp) and cell proliferation toxicity test. The content of propionic acid and butyric acid in the model was determined by gas chromatography mass spectrometry (GC-MS) before and after inoculating with Lactobacillus. [Results] The TEER of Caco-2 cell monolayer was 1 290.73 Ω·cm2 on the 11th day, 1 319.31 Ω·cm2 on the 15th day, and the Papp was less than 1×10?6 cm/s, the cells were closely connected and covered with a layer of microvilli perpendicular to the cell surface. The survival rates of Caco-2 cells were 99.03% and 91.42% after treating with 1 mmol/L propionic acid and 1 mmol/L butyric acid for 3 hours respectively. The propionic acid content of Caco-2 cells in the model inoculated with Lactobacillus plantarum P54, P58, P67, P97, P123 and P198 was higher than non-inoculate significantly (P<0.05), and the butyric acid content of Caco-2 cells in the model inoculated with Lactobacillus fermentum F146 and Lactobacillus plantarum P1 was higher than non-inoculate significantly (P<0.05). [Conclusion] The Lactobacillus strains used in this study can promote the absorption of propionic acid or butyric acid by Caco-2 cells in the model.