Study of endoplasmic reticulum stress in human dental pulp cells induced by oxygen-glucose deprivation culture in vitro
10.12016/j.issn.2096-1456.2022.04.003
- Author:
LI Lifen
1
;
ZHU Yaqin
1
;
JIANG Long
1
Author Information
1. Department of General Dentistry, Shanghai Ninth People’s Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine; National Center for Stomatology; National Clinical Research Center for Oral Disease; Shanghai key Laboratory of Stomatology
- Publication Type:Journal Article
- Keywords:
human dental pulp cells;
oxygen-glucose deprivation;
endoplasmic reticulum stress;
splicing x-box binding protein 1;
activating transcription factor 4;
C/EBP homologous protein;
RNA-activated protein kinase-like ER-resident kinase;
eukaryotic initiation factor-2α
- From:
Journal of Prevention and Treatment for Stomatological Diseases
2022;30(4):245-250
- CountryChina
- Language:Chinese
-
Abstract:
Objective:Oxygen-glucose deprivation (OGD) is used to mimic ischemia in vitro to observe whether endoplasmic reticulum (ER) stress is involved in human dental pulp cells (hDPCs) after OGD and to better understand the regulatory mechanism of hDPCs in ischemia.
Methods: hDPCs were cultured in glucose-free DMEM and hypoxia (volume fraction 2% O2) to establish an hDPCs OGD model in vitro, which mimics hDPCs ischemia in vitro. hDPCs were divided into a control group (normal culture) and an experimental group (OGD 0 h, 2 h, 4 h and 8 h groups). After pretreatment with OGD for 0, 2, 4 and 8 h, hDPC viability was measured by methylthiazol tetrazolium (MTT) assay. qRT-PCR was used to detect the mRNA expression of ER stress markers [splicing x-box binding protein1 (sXBP1), activating transcription Factor 4 (ATF4) and C/EBP homologous protein (chop)]. Western blot was used to detect the protein expression of ER stress markers [phosphorylated RNA-activated protein kinase-like ER-resident kinase (p-perk) and phosphorylated eukaryotic initiation factor-2α (p-eIF2α)].
Results:Compared with OGD 0 h group, cell viability of hDPCs decreased when exposed to OGD treatment for 2 h, 4 h and 8 h. Compared with the control group, mRNA expressions of ER stress makers (sXBP1, ATF4 and chop) and the protein expressions of ER stress protein markers (p-perk andp-eIF2α) increased in OGD treatment cells after 4 h were higher in OGD cells. The differences were statistically significant (P<0.05).
Conclusion:The results indicate that ER stress response is involved in hDPCs in OGD treatment.
- Full text:体外氧糖剥夺培养条件促进人牙髓细胞内质网应激的研究.pdf
