5-HT Promotes Proliferation and Inhibits Apoptosis of Megakarycytes through 5-HT2BR
10.19746/j.cnki.issn1009-2137.2025.01.011
- VernacularTitle:5-HT通过5-HT2BR促进巨核细胞生长和抗巨核细胞凋亡
- Author:
Hui-Min KONG
1
;
Yu-Rong CEN
;
Mo YANG
;
Qiang PENG
;
Jin-Qi HUANG
Author Information
1. 中山大学附属第七医院输血科,广东 深圳 518107
- Publication Type:Journal Article
- Keywords:
5-HT;
Meg-01 cells;
5-HT2BR;
proliferation;
apoptosis
- From:
Journal of Experimental Hematology
2025;33(1):75-81
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of 5-hydroxytryptamine(5-HT)on the proliferation,apoptosis and colony-forming unit-megakaryocyte(CFU-MK)of Meg-01 cells and its possible mechanisms.Methods:The uptake and metabolism of 5-HT in Meg-01 cells were analysed by reverse-phase high-performance liquid chromatography(RP-HPLC)with electrochemical detection.The expression of 5-HT2B receptor(5-HT2BR)in megakaryocytes was detected by immunofluorescence staining.The cell proliferation and viability were measured by MTT and Trypan blue staining after Meg-01 cells were single-cultured or co-cultured with different concentrations of 5-HT/5-HT2BR inhibitor Ketanserin for 48 h.Meg-01 cells were incubated with 5-HT/Ketanserin for 72 h,then the flow cytometry was used to detect early apoptosis of the cells and the activity of caspase-3.Using CFU-MK assay to investigate the effect of 5-HT on the differentiation of megakaryocytes.Results:5-HT could be uptaken by Meg-01 cells,and metabolized into 5-hydroxyindoleacetic acid(5-HIAA).The expression of 5-HT2BR on megakaryocytes could be detected after immunofluorescence staining.5-HT could promote the proliferation of Meg-01 cells at a dose-dependent manner(r=0.82),with the most significant effect observed at a concentration of 200 nmol/L(P＜0.001).Trypan blue staining also indicated that 200 nmol/L 5-HT had the most significant effect on the viability of Meg-01 cells(P＜0.05).The proliferation of Meg-01 cells treated with 5-HT was increased compared with the untreated control(P＜0.001),while the combination of 5-HT with ketanserin downregulated this effect.5-HT significantly reduced the early apoptosis rate(P＜0.001)and caspase-3 activity(P＜0.05)of Meg-01 cells,while addition of ketanserin significantly increased the early apoptosis rate of Meg-01 cells(P＜0.001)and caspase-3 activity also increased to some extent.5-HT promoted the formation of CFU-MK in bone marrow cells in a dose-dependent manner(r=0.89).The addition of ketanserin reduced the promoting effect of 5-HT on CFU-MK formation(P＜0.01).Conclusion:There may be monoamine oxidase present in megakaryocytes,which can metabolize and decompose 5-HT into 5-HIAA.5-HT may promote the proliferation and differentiation of megakaryocytes through 5-HT2BR.Besides,5-HT can also reduce the apoptosis of megakaryocytes,and its anti-apoptotic effect may be mediated by 5-HT2BR and caspase-3 pathways.
