Jianpi Qingchang Decoctioncontaining serum regulates the autophagy of interstitial cells

Objective: To investigate the effects of Jianpi Qingchang Decoctioncontaining serum (JQDCS) on interstitial cells of Cajal (ICCs) autophagy and cell cycle arrest in vitro. Methods: ICCs were collected from the small intestines of mice and analyzed using an anticKit antibody. ICCs were divided into five groups:the blank group, the rapamycin group (an autophagy inducer), the 5% (rapamycin + 5% JQDCS), the 20% (rapamycin + 20% JQDCS) JQDCS groups, and the 3Methyladenine (3MA) group (rapamycin + 3MA; positive control). Transmission electron microscopy was used to observe the ultrastructure of ICCs. Western blotting was used to detect the expression of microtubuleassociated protein 1 light chain 3 (LC3Ⅱ), Beclin1, phosphatidylinositol 3kinase (PI3K), pPI3K, protein kinase B (AKt), pAKt, mammalian target of rapamycin (mTOR), and pmTOR. Ca2+ current was examined by patchclamp experiments. Cell cycle was detected by flow cytometry. Results: Unlike in the rapamycin group, the ICC structures were more integrated and a lower number of autophagic vacuoles were observed in the 20% JQDCS group. Moreover, the expression of LC3Ⅱ and Beclin1 decreased, the expression of cKit, pPI3K, pAKt increased, the maximum current density value decreased, and the number of cells in the G1 phase increased while those in the G2/M phase decreased, with the addition of 20% JQDCS. Conclusion: JQDCS can antagonize rapamycininduced autophagy in ICCs in vitro by promoting the phosphorylation of PI3K/AKt pathway, inhibiting Ca2+ inflow, and regulating the cell cycle.