Effect of Nrf2 overexpression on anti-hypoxia and anti-apoptotic ability of mesenchymal stem cells
10.3969/j.issn.1674-7445.2018.02.004
- VernacularTitle:核因子E2相关因子2过表达对间充质干细胞抗缺氧和抗凋亡能力的影响
- Author:
Rongqiang LIU
1
;
Zenan YUAN
;
Xiaocai WU
;
Wei LIU
;
Guoying WANG
Author Information
1. 中山大学附属第三医院肝脏外科暨肝移植中心
- Keywords:
Nuclear factor erythroid-2-related factor 2(Nrf2);
Mesenchymal stem cells(MSCs);
Lentiviral vector;
Transfection;
Plasmid;
Apoptosis;
Hypoxia;
Flow cytometry;
Human embryonic kidney cell;
Ischemia-reperfusion injury (IRI)
- From:
Organ Transplantation
2018;9(2):110-115
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of nuclear factor erythroid-2-related factor 2(Nrf2) on the anti-hypoxia and anti-apoptotic ability of mesenchymal stem cells(MSCs). Methods Human embryonic kidney cells(293FT) were transfected with recombinant plasmid which overexpressed Nrf2 and helper plasmid. High-titer lentivirus which overexpressed Nrf2 were obtained. MSCs were transfected with lentivirus with Nrf2 overexpression and empty lentiviral vector to establish Nrf2-MSCs which stably overexpressed Nrf2 (Nrf2 overexpression group) and green fluorescent protein (GFP)-MSCs(control group). The expression of green fluorescent in 2 groups was observed by fluorescence microscope. The expression level of Nrf2 protein in 2 groups was measured by Western Blot. The anti-hypoxia ability of 2 groups was observed by light microscope. The anti-apoptotic ability of 2 groups was measured by flow cytometry. Results Nrf2-MSCs which stably overexpressed Nrf2 were successfully established. Western Blot analysis revealed that the expression level of Nrf2 protein in the Nrf2 overexpression group was significantly higher than that in the control group(P<0.01). After 15 h hypoxia treatment, the cell activity in the Nrf2 overexpression group was significantly higher than that in the control group. Flow cytometry showed that the apoptosis rate in the Nrf2 overexpression group was (30.9±1.4)%, significantly lower than (61.3±1.3)% in the control group(P<0.05). Conclusions Nrf2-MSCs which can stably overexpress Nrf2 possess certain anti-hypoxia and anti-apoptotic ability in hypoxia environment.
