Effect of Conditioned Medium of Bone Marrow Mesenchymal Stem Cells Pretreated by Ghrelin on Proliferation, Migration and Apoptosis of Endothelial Cells
- VernacularTitle:Ghrelin 预处理骨髓间充质干细胞来源上清液对内皮细胞增殖、迁移及凋亡的影响
- Author:
Shan⁃hui GE
1
;
Li⁃shan ZHANG
1
;
Shan LIN
1
;
Mian ZENG
1
Author Information
1. Medical Intensive Care Unit, First Affiliated Hospital, Guangzhou 510080, China
- Publication Type:Journal Article
- Keywords:
acute respiratory distress syndrome;
ghrelin;
bone marrow mesenchymal stem cells;
endothelial cell
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2020;41(5):681-689
- CountryChina
- Language:Chinese
-
Abstract:
【Objective】 To investigate the impact of conditioned medium derived from bone marrow mesenchymal stem cells(BMSCs) pretreated by ghrelin on proliferation, migration and apoptosis of endothelial cells(EA.hy926) and its underlying mechanism. 【Methods】 Rat BMSCs were isolated and cultured in vitro by the whole bone marrow adherence method, and surface markers of BMSCs were identified with flow cytometry. The conditioned medium of BMSCs and BMSCs pretreated by high, moderate and low concentration(100 nmol/L, 10 nmol/L and 1 nmol/L) of ghrelin for 24 hours was collected and co⁃cultured with EA.hy926 for 24 hours, and the serum⁃free medium was considered as the control group. Cell viability was detected by cell counting kit⁃8 while migration ability was analyzed by scratch assays. The cell apoptosis rates were measured by Annexin V method after treatment of lipopolysaccharide for 24 hours, and the protein expression of bax, caspase3, and β⁃catenin were detected by western blot. 【Results】 Compared with untreated BMSCs group, high concentration of ghrelin pretreated BMSCs group indicated higher cell viability and migration ability(P< 0.05). In light of the result of flow cytometry, endothelial cells apoptosis rate of 100 nmol/L ghrelin pretreated BMSCs group(34.51±3.51%) were remarkably lower than that of untreated BMSCsgroup(P<0.05). As revealed by western blot, expression level of bax, caspase3, and β⁃catenin were reduced in 100 nmol/L ghrelin pretreated BMSCs group compared with untreated BMSCs group(P<0.05) . 【Conclusion】 Endothelial cell function was improved when co⁃cultured in conditioned medium of BMSCs pretreated by ghrelin, and the attenuation of LPS⁃induced apoptosis was related with inhibition of Wnt/β⁃catenin signal path in endothelial cells.
