Inhibitory effect of exosomes secreted by human umbilical cord mesenchymal stem cells on apoptosis of oxygen-glucose deprived reoxygenation model of venous endothelial cells
10.3760/cma.j.issn.1674-6554.2019.12.001
- VernacularTitle: 人脐带间充质干细胞源外泌体对氧糖剥夺复氧静脉内皮细胞凋亡的抑制作用
- Author:
Yichao YE
1
,
2
;
Xiaohong LI
3
;
Xinyu SHI
4
;
Zhenwen ZHANG
4
;
Xiaoyin LIU
4
;
Jian CHEN
1
;
Zhe ZHANG
1
;
Weizhou WU
1
;
Jingjing WANG
4
;
Hongxia ZHOU
5
;
Yi WANG
5
Author Information
1. Logistics University of People's Armed Police Forces, Tianjin 300309, China
2. Institution of Brain Trauma and Neurology Disease of People's Armed Police Forces, Tianjin Key Laboratory of Neurotrauma Repair, Tianjin 300162, China
3. Academy of Medical Engineering and Translational Medicine, Tianjin Uniersity, Tianjin 300072, China
4. Institution of Brain Trauma and Neurology Disease of People's Armed Police Forces, Tianjin Key Laboratory of Neurotrauma Repair, Tianjin 300162, China
5. Department of Neurology, Tianjin Hospital, Tianjin 300211, China
- Publication Type:Journal Article
- Keywords:
Human umbilical mesenchymal stem cells;
Exosomes;
Oxygen-glucose deprivation reoxygenation;
Human umbilical venous endothelial cells;
Apoptosis
- From:
Chinese Journal of Behavioral Medicine and Brain Science
2019;28(12):1057-1063
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the inhibitory effect of exosomes secreted by human umbilical cord mesenchymal stem cells(HUCMSC) on apoptosis of human umbilical vein endothelial cells(HUVEC) after model group(oxygen-glucose deprivation reoxygenation), and to clarify its possible mechanism.
Methods:Human umbilical cord mesenchymal stem cells were cultured. The collected cell supernatant was stored in a centrifugal tube. The exosomes secreted by human umbilical cord mesenchymal stem cells were extracted by ultracentrifugation and identified. Human umbilical vein endothelial cells were randomly divided into control group, model group and different concentrations of HUCMSC-EXO(20 μg/ml, 40 μg/ml, 60 μg/ml) treatment groups(adding HUCMSC-EXO into the model group) . The morphological changes of HUVEC cells in each group were observed by inverted phase contrast microscope, and the proliferation inhibition rate of HUVEC in each group was measured by CCK-8 reagent. Western blot was used to detect the expression of apoptosis-related proteins Caspase-3, Bax, Bcl-2 and hypoxia-associated protein hypoxia inducible factor 1α(HIF-1α). Inhibitor(HIF-1α inhibitor) + model group and HUCMSC-EXO + inhibitor + model group were added on the basis of the above experiments. Western blot analysis was performed to observe the effects of HUCMSC-EXO, inhibitor and both of them on HIF-1α and Bax expressions in HUVEC.
Results:HUCMSC-EXO was successfully extracted and identified. Compared with the control group, the volume of HUVEC in the model group and the HUCMSC-EXO group with different concentrations decreased, became round, connected and evacuated, and the growth state was poor under the inverted phase contrast microscope.CCK-8 detection showed that the cell viability in the HUCMSC-EXO group was significantly higher than that in the model group, the difference was statistically significant (t=9.23, P<0.05). Western blot analysis showed that compared with the control group, the expression levels of Caspase-3 ((0.296±0.038), (0.879±0.088); t=14.92, P<0.05), Bax((0.234±0.034), (0.762±0.084); t=14.36, P<0.05) of HUVEC in the model group were up-regulated, and the expression level of Bcl-2 was down-regulated ((0.863±0.103), (0.387±0.059); t=9.85, P<0.05), with statistically significant differences. Compared with the model group, the expression levels of Caspase-3( (0.586±0.075); t=6.24, P<0.05), Bax((0.311±0.055); t=11.01, P<0.05) and Bcl-2((0.665±0.071); t=7.45, P<0.05) of HUVEC in the HUCMSC-EXO treatment group were down-regulated and the differences were statistically significant. Inhibitor intervention experiments showed that there were no significant differences between the inhibitor+ model group and HUCMSC-EXO+ inhibitor+ model group in the expression of HIF-1α protein ((0.348±0.055), (0.388±0.077); t=1.04, P>0.05)and Bax protein ((0.363±0.069), (0.370±0.064); t=0.18, P>0.05). But both of them were down-regulated compared with the model group (HIF-1α protein (0.919±0.064), Bax protein (0.902±0.071)), the differences were significant( t=13.56, t=13.03, both P<0.05).
Conclusion:HUCMSC-EXO has a protective effect on OGD/R model of HUVEC, and its mechanism may be related to the down-regulation of HIF-1α expression.
