Inducible nitric oxide synthase inhibitor 1 400W suppresses endoplasmic reticulum stress and alleviates ischemia-reperfusion injury in human intrahepatic bile duct epithelial cells
10.3760/cma.j.issn.0254-1785.2019.04.011
- VernacularTitle:1400W抑制内质网应激减轻人肝内胆管上皮细胞缺血再灌注损伤
- Author:
Qiwen YU
1
;
Hongwei TANG
;
Dongjing YANG
;
Wenzhi GUO
;
Jie LI
;
Shuijun ZHANG
Author Information
1. 郑州大学第一附属医院肝胆胰外科河南省器官移植医学工程技术中心郑州市肝胆胰疾病与器官移植医学重点实验室郑州市器官移植技术与应用工程重点实验室
- Keywords:
Ischemia-reperfusion injury;
Bile duct epithelial cells;
Nitric oxide synthase
- From:
Chinese Journal of Organ Transplantation
2019;40(4):241-244
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the role and mechanism of inducible nitric oxide synthase inhibitor 1 400W in alleviating ischemia-reperfusion injury of human intrahepatic bile duct epithelial cells.Methods Human intrahepatic bile duct epithelial cells (HIBEC) in logarithmic phase were inoculated into culture plate at an appropriate density.The samples were randomly divided into control group (group C),ischemiareperfusion group (group I/R) and ischemia-reperfusion + 1 400W group (group I/R + 1 400W).Group C was cultured routinely;cells in I/R and I/R + 1 400W groups were placed in a three-gas incubator for 12h for simulating ischemia and then normal culture for 6h for simulating reperfusion.The I/R + 1 400W group had a final concentration of 100 μmol/L of 1 400W before ischemia and hypoxia.After reperfusion,cells and culture medium were collected,CCK 8 was used for detecting cell vitality,microplate method for detecting the content of lactate dehydrogenase (LDH) in culture medium,AnnexinV-FITC/PI double stain for detecting apoptosis level,Western blot for analyzing the expressions of endoplasmic reticulum stress (ERS)related protein cysteinyl aspartic acid protease 12 (caspase-12),glucose regulatory protein 78 (GRP78) C/EBP homologous protein (CHOP) and inducible nitric oxide synthase (iNOS).Results As compared with group C,cell viability significantly decreased in I/R and I/R+ 1 400W groups (53.8％ ± 2.3％ vs.100％,66.5 ％ ± 2.8 ％ vs.100 ％) (P＜0.05) while LDH increased markedly in cell culture medium (287.4 ±9.0U/L vs 120.2 ± 8.7U/L,212.0 ± 8.3U/L vs 120.2 ± 8.7U/L) (P＜0.05).Apoptosis accelerated markedly (41.5％±2.3％ vs5.2％±0.5％,32.7％± 1.8％ vs 5.2％±0.5％) (P＜0.05) and the expressions of caspase-12,GRP78,CHOP and iNOS spiked (P＜0.05);as compared with I/R group,cell viability of I/R+ 1 400W group rose while LDH,apoptosis level,caspase-12,GRP78 and CHOP declined in cell culture medium (P＜0.05).Conclusions 1 400W may alleviate ischemia-reperfusion injury of human intrahepatic bile duct epithelial cells and its mechanism may be correlated with a suppression of endoplasrnic reticulum stress.
