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Objective: To investigate the effects of Aurantii Fructus lmmaturus (Zhishi,ZS) and Atractylodis Macrocephalae Rhizoma (Baizhu,BZ)-containing serum on glutamate-induced autophagy in rat colonic interstitial cells of Cajal (ICCs) and to analyze the underlying mechanism.Methods: Rat colonic ICCs cultured in vitro were identified by fluorescence and then stimulated with glutamic acid (5 mmol/L) for 24 h to establish a cell model of autophagy.The cells were then treated with different concentrations of ZSBZ-containing serum or rat serum.The viability of the ICCs was detected with cell counting kit-8 assays,and cell apoptosis rates were examined with flow cytometry.The ultrastructure and autophagosomes in the ICCs were observed using transmission electron microscopy.The effects of ZSBZ-containing serum on apoptosis-associated mediators were assessed by Western blotting and real-time quantitative polymerase chain reaction.In addition,microtubule-associated protein light chain 3 (LC3),p-phosphoinositide 3-kinase (p-PI3K),p-Akt and p-mammalian target of rapamycin (pmTOR) expression was detected via Western blotting analysis.Results: Compared to those in the model group,ICC viability and apoptosis rates were significantly increased by ZSBZ-containing serum (P ＜ 0.05).In addition,the expression levels of Beclin-1,LC3,p-PI3K,p-Akt and p-mTOR were significantly lower (P ＜ 0.05) and Bcl-2 expression was higher in the ZSBZ-containing serum treatment groups than in the model group (P ＜ 0.05).Conclusion: Our findings demonstrated that ZSBZ protects glutamic acid-stimulated ICCs,and this beneficial effect may be mediated by a reduction in autophagy via inhibition of the PI3K/Akt/mTOR pathway.