Inflammatory periodontal stem cells mediate interleukin-1β secretion of macrophage by regulating macrophage endoplasmic reticulum stress
10.3760/cma.j.cn112144-20201105-00553
- VernacularTitle:炎性牙周膜干细胞通过调控巨噬细胞内质网应激介导白介素-1β分泌的研究
- Author:
Yang ZHANG
1
;
Yazheng WANG
;
Dongdong FEI
;
Xige ZHANG
;
Zixuan LIAO
;
Lingxia LIU
;
Qintao WANG
Author Information
1. 第四军医大学口腔医学院牙周病科 军事口腔医学国家重点实验室 口腔疾病国家临床医学研究中心 陕西省口腔生物工程技术研究中心，西安 710032
- Keywords:
Periodontitis;
Macrophages;
Periodontal ligament stem cells;
Endoplasmic reticulum stress;
Interleukin-1β;
Inflammatory environment
- From:
Chinese Journal of Stomatology
2021;56(4):329-334
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect and mechanism of periodontal ligament stem cell (PDLSC) from inflammatory environment on the secretion of interleukin-1β (IL-1β) by macrophages.Methods:PDLSCs were pretreated with lipopolysaccharide (LPS) in order to simulate the inflammatory environment. Human monocyte cell line (THP-1) cells were treated with conditioned media collected from healthy and inflammatory PDLSCs respectively and divided into conditioned medium of health PDLSC (CM-H) group and conditioned medium of LPS-PDLSC (CM-LPS) group. After 24 h of co-culture, the condition media were abandoned and THP-1 cells were then cultured for another 24 h. The expression of IL-1β in THP-1 cells supernatant was detected by enzyme-linked immunosorbent assay (ELISA). Quantitative real time-PCR (qRT-PCR) was used to detect the expression of glucose regulated protein 78 (GRP78), activating transcription factor-6 (ATF6), inositol requiring enzyme 1 (IRE1), protein kinase R-like endoplasmic reticulum kinase (PERK), CCAAT enhancer binding protein homologous protein (CHOP), activating transcription factor-4 (ATF4) and X box binding protein 1 spliced (XBP1s), which were all related with endoplasmic reticulum stress (ERS), in THP-1 cells. The expressions of proteins GRP78 and CHOP were detected by Western blotting. Furthermore, THP-1 cells, which pretreated with ER inhibitor 4-phenylbutyrate (4-PBA) for intervention experiments were grouped by various concentrations of 4-PBA including groups 0 (control group), 1, 10 and 20 mmol/L and treated with condition medium of inflammatory PDLSC. ELISA was used to detect IL-1β expression and qRT-PCR to detect expression of ERS related genes.Results:ELISA results showed that the expression of IL-1β in THP-1 cells of group CM-LPS [(31.35±2.11) ng/L] was significantly higher than group CM-H [(8.19±1.51) ng/L] ( t=12.60, P<0.01). qRT-PCR results showed that the relative expressions of GRP78, ATF6, IRE1, PERK, CHOP, ATF4 and XBP1s genes in THP-1 cells of group CM-LPS (1.782±0.070, 1.387±0.204, 1.404±0.119, 1.777±0.187, 1.325±0.156, 1.295±0.066 and 1.137±0.149, respectively) were significantly higher than those in group CM-H ( P<0.05). In the 4-PBA intervention experiment, compared with group 0 mmol/L, the expressions of GRP78, IRE-1, ATF-6, PERK and CHOP were significantly lower in group 1, 10 and 20 mmol/L ( P<0.05). Moreover, compared with control group [(31.23±1.98) ng/L], the expression of IL-1β in THP-1 cells were significantly lower in group 10 mmol/L [(21.20±0.37) ng/L] and group 20 mmol/L [(23.85±1.80) ng/L] ( P<0.05) with ERS inhibited. Conclusions:PDLSC from inflammatory environment could promote IL-1β secretion of macrophages through upregulating macrophages ERS.
