Protective effect of carbachol postconditioning on hypoxia/reoxygenation-induced injury in human gastric epithelial cells

• Published in: Life sciences (1973) Vol. 144; pp. 234 - 242
• Main Authors: Han, Hongxia, Yang, Jun, Fei, Sujuan, Liu, Zhangbo, Zhu, Shengping, Dong, Qiuju, Gao, Zhifeng, Wang, Shihui, Zhang, Jianfu
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Inc 01.01.2016
• Subjects: Apoptosis - drug effects; Calcitonin Gene-Related Peptide - biosynthesis; Carbachol - pharmacology; Carbachol postconditioning (CAR-P); Cell Line; Cell Survival; Cholinergic muscarinic receptor; Epithelial Cells - drug effects; Gastric Mucosa - pathology; Human gastric epithelial cells (hGES-1); Humans; Hypoxia - pathology; Hypoxia/reoxygenation (H/R); Ischemic Postconditioning; MAP Kinase Signaling System - drug effects; Muscarinic Agonists - pharmacology; Receptors, Muscarinic - drug effects; Stomach - drug effects; Stomach - pathology; Cholinergic muscarinic receptor; Human gastric epithelial cells (hGES-1); Carbachol postconditioning (CAR-P); Hypoxia/reoxygenation (H/R)
• ISSN: 0024-3205; 1879-0631
• DOI: 10.1016/j.lfs.2015.11.029

We investigated the protective effects of carbachol postconditioning (CAR-P) on acute gastric mucosal injury induced by hypoxia/reoxygenation (H/R) and its possible mechanisms. Cell viability was detected by methyl thiazolyl tetrazolium (MTT). The apoptotic cells were examined by Hoechst 33258 staining. Flow cytometric analysis, lactate dehydrogenate (LDH) release assay, immunocytochemistry, and western blotting were used to investigate the effects of CAR-P on acute gastric mucosal injury induced by H/R. The model of H/R was established by hypoxia induction(94% N2+1% O2+5% CO2 for 2h) and reoxygenation (normoxic condition for 4h, 8h and 16h). Our study observed the protective effect of carbachol postconditioning on H/R-induced injury in human gastric epithelial cell lines (hGES-1) cells, which is achieved by direct activation of vanilloid receptor subtype 1 (VR1) and production of calcitonin gene-related peptide (CGRP), and in the inhibition of cell apoptosis. In the study, we demonstrate that CAR-P has protective effects on the H/R-induced injury in hGES-1 cells, and these effects are associated with cholinergic muscarinic receptors (CMR), VR1, and extracellular signal-regulated kinase (ERK) signaling pathway. Our findings might provide a new and improved understanding of CAR-P function and an effective treatment strategy for acute gastric mucosal injury induced by H/R.