Role of the GRP78-c-Src signaling pathway on osteoblast differentiation of periodontal ligament fibroblasts in-duced by cyclic mechanical stretch
10.7518/hxkq.2024.2023354
- VernacularTitle:GRP78-c-Src信号通路介导周期性牵张力作用下牙周膜成纤维细胞成骨分化的机制研究
- Author:
Jing HU
1
;
Zhihua CUI
;
Keqiang HUANG
;
Rongjian SU
;
Song ZHAO
Author Information
1. 锦州医科大学生命科学研究院公共实验平台,锦州 121001
- Keywords:
cyclic mechanical stretch;
osteogenic differentiation;
periodontal ligament fibroblasts;
glucose regu-lated protein 78
- From:
West China Journal of Stomatology
2024;42(3):304-312
- CountryChina
- Language:Chinese
-
Abstract:
Objective This study aims to investigate the influence of glucose regulated protein(GRP)78 on osteo-blast differentiation in periodontal ligament fibroblasts(PDLFs)under cyclic mechanical stretch and determine the under-lying mechanism.Methods FlexCell 5000 cell mechani-cal device was applied to simulate the stress environment of orthodontic teeth.GRP78High and GRP78Low sub-population were obtained by flow sorting.Gene transfec-tion was performed to knockdown GRP78 and c-Src ex-pression and overexpress c-Src.Western blot analysis was used to detect the protein expression of Runt-related gene 2(RUNX2),Osterix,osteocalcin(OCN),and osteopontin(OPN).Immunoprecipitation assay was used to determine the interaction of GRP78 with c-Src.The formation of cellular mineralized nodules was determined by alizarin red staining.Results GRP78 was heterogeneously expressed in PDLFs,and GRP78High and GRP78Low subpopulations were obtained by flow sorting.The osteogenic differentiation ability and phosphorylation level of c-Src kinase in the GRP78High subpopulation were significantly increased com-pared with those in GRP78Low subpopulation after cyclic mechanical stretch(P<0.05).GRP78 interacted with c-Src in PDLFs.The overexpression c-Src group showed significantly increased osteogenic differentiation ability than the vector group(P<0.05),and the sic-Src group showed significantly decreased osteogenic differentiation ability(P<0.05)after cy-clic mechanical stretch.Conclusion GRP78 upregulates c-Src expression by interacting with c-Src kinase and pro-motes osteogenic differentiation under cyclic mechanical stretch in PDLFs.
