Antophagy inhibition can decrease AR42J cell apoptosis induced by exendin-4
10.3760/cma.j.issn.1007-8118.2018.12.013
- VernacularTitle:抑制自噬对艾塞那肽诱导的AR42J细胞凋亡的影响
- Author:
Weidong ZHU
1
;
Xiao YU
;
Xia LI
;
Can YU
;
Hongwei ZHU
;
Duo HAN
;
Hui HUANG
;
Zhiqiang LI
Author Information
1. 湖南省岳阳市二人民医院肝胆胰外科
- Keywords:
Exendin-4;
Pancreatic acinar cell;
Apoptosis;
Autophagy inhibitor,3-MA
- From:
Chinese Journal of Hepatobiliary Surgery
2018;24(12):848-852
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore whether exendin-4 inhibits AR42J cells and its mechanism.Methods AR42J cells were treated with exendin-4 under multiple concentrations(1,5,10 pmol/L) at 24,48,72,96,120 h to assess its cell viability by MTT assay and got the IC-50 and time points.Then checking whether exendin-4 could induce the AR42Jcells apoptosis by setting normal control (NC) group,exendin-4 (Ex-4) group and Ex-4+ z-VAD-fmk (apoptosis inhibitor) group,and exploring whether 3-MA which is autophagy inhibitor could inhibit the AR42J cells apoptosis induced by exendin-4 by setting NC group,Ex-4 group and Ex-4+3-MA group.Cell viability was analyzed by MTT and the cells apoptosis was detected by flow cytometry and the protein levels of caspase-3,LC3 and p62 were studied by Western blot.Results Concentration of 10 pmol/L exendin-4 and and time point 72 h were selected for the further study.z-VAD-fmk pretreatment can significantly inhibit the cell viability of exendin-4 by (81.2±3.3)％ vs.(49.4±3.0)％ (P＜0.05).Flow cytometry showed that exendin-4 could induce the AR42J cells apoptosis by (28.2± 1.4)％ vs.(3.6±0.8)％,and increased the caspase-3 level by Western blot,which both can be reversed by (79.1 ±2.3) ％ vs.(49.8±2.5)％ (P＜0.05) when the cells were treated for 72 h,as was apoptosis ratio by (14.5±2.1)％ vs.(29.2±3.2)％.Western blot showed that exendin-4 can upregulate protein levels of LC3B-Ⅱ,p62 和 caspase-3 and 3-MA,and pretreatment can inhibit the upregulation of LC3B-Ⅱ and caspase-3 but further increased the upregulation of p62 induced by exendin-4.Conclusions Exendin-4 can induce AR42J cells apoptosis and 3-MA pretreating can inhibit exendin-4 cytotoxicity through downregulating autophagy.So auto phagy inhibitor 3-MA could potentially extenuate the cytotoxicity of exendin-4 in pancreatic acinar cells.
