Protection of ghrelin postconditioning on hypoxia/ reoxygenation in gastric epithelial cells  

作　　者：Zhang-Bo Liu Su-Juan Fei Sheng-Ping Zhu Jin-Zhou Zhu Hong-Xia Han Qiu-Ju Dong Jian-Fu Zhang 

机构地区：[1]DepartmentofGastroenterol-ogy,TheAffiliatedHospitalofXuzhouMedicalCollege,99WestHuaihaiRoad,Xuzhou221002,JiangsuProvince,China [2]Department of Physiology, Xuzhou MedicalCollege, Xuzhou 221002, Jiangsu Province, China

出　　处：《World Journal of Gastroenterology》2012年第38期5377-5388,共12页世界胃肠病学杂志（英文版）

基　　金：Supported by National Natural Science Foundation of China, No.30570671;the Educational Department Science Research Foundation of Jiangsu Province, No. 99KJB310005 and 05KJB310134

摘　　要：AIM: To investigate the protective effect and mechanisms of ghrelin postconditioning against hypoxia/reoxygenation (H/R)-induced injury in human gastric epithelial cells. METHODS: The model of H/R injury was established in gastric epithelial cell line (GES-1) human gastric epithelial cells. Cells were divided into seven groups: normal control group (N); H/R postconditioning group; DMSO postconditioning group (DM); ghrelin postconditioning group (GH); D-Lys3-GHRP-6 + ghrelin postconditioning group (D + GH); capsazepine + ghrelin postconditioning group (C + GH); and LY294002 + ghrelin postconditioning group (L + GH). 3-(4,5-dimethylthazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to detect GES-1 cell viability. Hoechst 33258 fluorochrome staining and flow cytometry were conducted to determine apoptosis of GES-1cells. Spectrophotometry was performed to determine release of lactate dehydrogenate (LDH). Protein expression of Bcl-2, Bax, Akt, and glycogen synthase kinase (GSK)-3β was determined by western blotting. Expression of vanilloid receptor subtype 1 (VR1), Akt and GSK-3β was observed by immunocytochemistry. RESULTS: Compared with the H/R group, cell viability of the GH group was significantly increased in a dosedependent manner (55.9% ± 10.0% vs 69.6% ± 9.6%, 71.9% ± 17.4%, and 76.3% ± 13.3%). Compared with the H/R group, the percentage of apoptotic cells in the GH group significantly decreased (12.38% ± 1.51% vs 6.88% ± 0.87%). Compared with the GH group, the percentage of apoptotic cells in the D + GH group, C + GH group and L + GH groups significantly increased (11.70% ± 0.88%, 11.93% ± 0.96%, 10.20% ± 1.05% vs 6.88% ± 0.87%). There were no significant differences in the percentage of apoptotic cells between the H/R and DM groups (12.38% ± 1.51% vs 13.00% ± 1.13%). There was a significant decrease in LDH release following ghrelin postconditioning compared with the H/R group (561.58 ± 64.01 U/L vs 1062.45 ± 105.29 U/L). There was a significant increase in LDH release in the DAIM： To investigate the protective effect and mechanisms of ghrelin postconditioning against hypoxia/reoxygenation （H/R）-induced injury in human gastric epithelial cells. METHODS： The model of H/R injury was established in gastric epithelial cell line （GES-1） human gastric epithelial cells. Cells were divided into seven groups： normal control group （N）; H/R postconditioning group; DMSO postconditioning group （DM）; ghrelin postconditioning group （GH）; D-Lys3-GHRP-6 ＋ ghrelin postconditioning group （D ＋ GH）; capsazepine ＋ ghrelin postconditioning group （C ＋ GH）; and LY294002 ＋ ghrelin postconditioning group （L ＋ GH）. 3-（4,5-dimethylthazol-2-yl）-2,5-diphenyl tetrazolium bromide （MTT） assay was used to detect GES-1 cell viability. Hoechst 33258 fluorochrome staining and flow cytometry were conducted to determine apoptosis of GES-1cells. Spectrophotometry was performed to determine release of lactate dehydrogenate （LDH）. Protein expression of Bcl-2, Bax, Akt, and glycogen synthase kinase （GSK）-3β was determined by western blotting. Expression of vanilloid receptor subtype 1 （VR1）, Akt and GSK-3β was observed by immunocytochemistry. RESULTS： Compared with the H/R group, cell viability of the GH group was significantly increased in a dosedependent manner （55.9% ± 10.0% vs 69.6% ± 9.6%, 71.9% ± 17.4%, and 76.3% ± 13.3%）. Compared with the H/R group, the percentage of apoptotic cells in the GH group significantly decreased （12.38% ± 1.51% vs 6.88% ± 0.87%）. Compared with the GH group, the percentage of apoptotic cells in the D ＋ GH group, C ＋ GH group and L ＋ GH groups significantly increased （11.70% ± 0.88%, 11.93% ± 0.96%, 10.20% ± 1.05% vs 6.88% ± 0.87%）. There were no significant differences in the percentage of apoptotic cells between the H/R and DM groups （12.38% ± 1.51% vs 13.00% ± 1.13%）. There was a significant decrease in LDH release following ghrelin postconditioning compared with the H/R group （561.58

关 键 词：Protection of ghrelin postconditioning on hypoxia/ reoxygenation in gastric epithelial cells 

分 类 号：Q813.11[生物学—生物工程] S828.2[农业科学—畜牧学]