CPC derivedexosome protects cardiomyocytes from oxidative stress
10.3969/j.issn.1001-1978.2015.12.007
- VernacularTitle:心脏干细胞分泌的 exosome对氧化应激下心肌细胞的保护作用研究
- Author:
Jing XIAO
;
Yu PAN
;
Xiaohong LI
;
Lin JIANG
;
Xiangyu YANG
;
Yueheng WU
;
Heping LEI
;
Xiyong YU
- Publication Type:Journal Article
- Keywords:
cardiac progenitor cells;
exosome;
mag-netic cell sorting system;
paracrine effect;
oxidative stress;
apoptosis
- From:
Chinese Pharmacological Bulletin
2015;(12):1656-1660
- CountryChina
- Language:Chinese
-
Abstract:
Aim To explore the anti-apoptotic function of cardiac progenitor cells(CPCs)-derived exosome in vitro.Method CPCs were isolated from mouse heart using Magnetic Cell Sorting(MACS)system.Flow Cy-tometry(FC)determine the purity of stem cell surface antigen-1 positive(Sca-1 +)CPCs.Exosome was puri-fied from conditional medium,and confirmed by West-ern blot using CD63 as a marker,Nanoparticle Traffic-king Analysis(NTA)was used to detect the diameters and concentration of exosome.Then the cells were di-vided into control groups and CPC-exosome pre-protec-tion groups.H2 O2 was added into H9c2 cells to induce oxidative stress.Western blot was adopted to determine the expression of cleaved caspase-3.Results ① Im-munofluorescence showed that CPCs isolated by MACS were positively expressing Sca-1 protein;FC analysis showed that typical purity of Sca-1 +CPCs from the first
preparations was more than 95%.② WB demonstrated that CD63 of exosome isolated from CCMwas positively expressed,and NTA results showed that the diameters of exosome were (82.33 ±3.06)nm(n =3).Micro-scope detected PKH-26 labeled exosome appeared in the cytoplasma of H9c2 cells.③ Western blot showed the CPC-exosome pre-protection groups significantly down-regulated the levels of cleaved caspase-3 com-pared to the control groups(P <0.05).Conclusion CPC can secrete exosome which carries many important cargos,which can effectively gather in H9c2 cells. CPC-exosome can protect H9c2 cells from the oxidative stress induced by H2 O2 .Our results highlight a new perspective strategy for cardiac disease.
