Jianpi Qingchang Decoction-containing serum regulates the autophagy of interstitial cells
Journal Title: Traditional Medicine Research - Year 2018, Vol 3, Issue 4
Abstract
Objective:To investigate the effects of Jianpi Qingchang Decoction-containing serum (JQD-CS) 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 anti-c-Kit antibody. ICCs were divided into five groups: the blank group, the rapamycin group (an autophagy inducer), the 5% (rapamycin + 5% JQD-CS), the 20% (rapamycin + 20% JQD-CS) JQD-CS groups, and the 3-Methyladenine (3-MA) group (rapamycin + 3-MA; positive control). Transmission electron microscopy was used to observe the ultrastructure of ICCs. Western blotting was used to detect the expression of microtubule-associated protein 1 light chain 3 (LC3-II), Beclin-1, phosphatidylinositol 3-kinase (PI3K), p-PI3K, protein kinase B (AKt), p-AKt, mammalian target of rapamycin (mTOR), and p-mTOR. Ca2+ current was examined by patch-clamp 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% JQD-CS group. Moreover, the expression of LC3-II and Beclin-1 decreased, the expression of c-Kit, p-PI3K, p-AKt 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% JQD-CS. Conclusion:JQD-CS can antagonize rapamycin-induced autophagy in ICCs in vitro by promoting the phosphorylation of PI3K/AKt pathway, inhibiting Ca2 + inflow, and regulating the cell cycle.
Authors and Affiliations
Yan-Cheng Dai, Ya-Li Zhang, Lie Zheng, You-Lan Chen, Xuan Chen, De-Liang Chen, Zhi-Peng Tang
Keywords
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- EP ID EP45227
- DOI 10.12032/TMR201812078
- Views 288
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