Effects of hypoxia H9c2 exosome on proliferation,migration and tube formation of HUVEC
- VernacularTitle:缺氧H9c2来源外泌体对HUVEC增殖、迁移和成管能力的影响
- Author:
Rui YANG
1
;
Qiong WEI
;
Yikun SUN
;
Mengzhu ZHAO
;
Xu CHENG
;
Menghua LIU
;
Dongmei ZHANG
Author Information
- Keywords: exosomes; myocardial infarction; endothelial cells; hypoxia; angiogenesis
- From: Tianjin Medical Journal 2024;52(7):714-719
- CountryChina
- Language:Chinese
- Abstract: Objective To investigate the role of H9c2-derived exosomes in regulating angiogenesis in rat cardiomyocytes under hypoxia.Methods The hypoxia model of H9c2 cells was prepared by mixed gas method(the hypoxia model group),and the normal cultured cells were used as the control group.The exosomes secreted by the two groups of cells were extracted respectively.The concentration and particle size of exosomes were detected by nanoparticle tracking analysis.The morphology and size of exosomes were detected by transmission electron microscopy.Western blot assay was used to verify the exosome marker proteins.The hypoxia model of human umbilical vein endothelial cells(HUVEC)was established.HUVECs were incubated with H9c2 exosomes and divided into the normoxia group,the hypoxia group,the hypoxia+normal H9c2 exosomes(EXO-C)group and the hypoxia+hypoxia H9c2 exosomes(EXO-M)group.The proliferation,migration and tube formation of HUVECs were detected by CCK-8 method,cell scratch test and Matrigel in vitro three-dimensional forming test.Results The results of exosome identification showed that the particle concentration of H9c2 exosome samples was 1×107-1×1012 particles/mL and the particle size was 40-160 nm in the normoxia group and the hypoxia group.The morphological characteristics were spherical or saucer-like structure,uniform in size and complete in shape.Exosome marker proteins TSG101,CD63 and CD9 were expressed,and there was no expression of negative protein Calnexin.Compared with the normoxic group,the proliferation ability,migration area and migration rate of HUVEC were significantly decreased in the hypoxic group,and the length of tube,the number of branches and the number of nodes were decreased(P＜0.01).Compared with the hypoxia group,the proliferation ability of HUVEC cells was decreased,the migration area was decreased,the migration rate was decreased and the length and number of branches involved in tube formation were further decreased in the EXO-M group(P＜0.05).Compared with the EXO-C group,the proliferation ability of the EXO-M group decreased,the cell migration area decreased and the migration rate decreased(P＜0.01).Conclusion Exosomes derived from hypoxic H9c2 can inhibit the proliferation,migration and tube formation of HUVEC.
