Human umbilical cord blood mesenchymal stem cells conditioned media inhibits hypoxia-induced apoptosis in H9c2 cells by activation of the survival protein Akt |
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Authors: | XUHE GONG GUANGPU FAN HUI LIU SIJIA WANG RONGCHONG HUANG |
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Affiliation: | 1.Department of Cardiology, Beijing Friendship Hospital Affiliated to Capital Medical University, Beijing, 100190, China
2 Department of Cardiac Surgery, Peking University People’s Hospital, Beijing, 100190,
3 Emergency Department, Beijing Friendship Hospital Affiliated to Capital Medical University, Beijing, 100190, China |
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Abstract: | This work aimed to study the beneficial role of human umbilical cord blood-derived mesenchymal stem cellconditioned medium (MSC-CM) in hypoxia-induced apoptosis in H9c2 cardiomyoblasts, in which the serine/heroine
kinases (Akt) pathway would be involved. For this, CM was collected by culturing MSCs in serum-free DMEM
medium for 24 h, and paracrine factors were analyzed by protein chip. H9c2 cells were divided into the following
groups: control group, hypoxia group, MSC-CM intervention group (CM group), MSC-CM + Akt phosphorylation
inhibitor (LY294002) group (LY group). Apoptosis of the H9c2 cells was tested with chromatin dye Hoechst 33342
and FITC-conjugated Annexin V apoptosis detection kit by flow cytometer after a hypoxia/serum deprivation (H/SD)
for 24 h. The apoptosis-related proteins were evaluated by Western blot. MSC-CM displayed significantly elevated
levels of growth factors, anti-inflammatory, and anti-apoptosis cytokines. On Hoechst 33342 apoptosis staining, the
H9c2 cell morphology displayed a lower proportion of apoptosis in the CM group than those in the hypoxia group,
while apoptosis was increased in LY group. Flow cytometer analysis revealed the apoptosis ratio in the CM group was
lower than the hypoxia group (12.34 ± 2.00% vs. 21.73 ± 2.58%; p < 0.05), while the LY group was significantly higher
(22.54 ± 3.89%). Active caspase-3 expression was increased in hypoxia group than control group (p < 0.05), but
decreased in CM group (p < 0.01). Umbilical cord blood-derived mesenchymal stem cell-conditioned media secrete
multiple paracrine factors that are able to inhibit hypoxia-induced H9c2 cardiomyoblasts apoptosis, and in which the
activation of Akt phosphorylation is involved to achieve the protective effect. |
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Keywords: | Umbilical cord blood mesenchymal stem cells paracrine actions H9c2 cells anti-apoptosis caspase-3 Akt |
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