Overexpression of NKx2.5 gene affects the anti apoptotic ability of mesenchymal stem cells and cardiac function after myocardial infarction
10.3760/cma.j.cn101721-20230911-00068
- VernacularTitle:过表达NKx2.5基因对间充质干细胞抗凋亡能力及对心肌梗死后心功能的影响
- Author:
Fugang MAO
1
;
Xinxin WU
;
Xinhao CHEN
;
Si LI
;
Dan YAN
;
Zhiyuan XIAO
;
Jigang HE
Author Information
1. 云南省第一人民医院超声科，昆明 650000
- Keywords:
Bone marrow mesenchymal stem cells;
Nkx2.5;
Anti-apoptosis;
Myocardial infarction
- From:
Clinical Medicine of China
2024;40(3):191-196
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of overexpression of Nkx2.5 gene on the anti apoptotic ability of bone marrow mesenchymal stem cells (BMSCs) and cardiac function after myocardial infarction.Methods:A cell ischemia model was established by culturing cells under oxygen glucose deprivation/reoxygenat (OGD/R) conditions. The experiment was divided into four groups: bone marrow mesenchymal stem cells cultured under normal conditions (BMSC group), BMSC group cultured under glucose and oxygen deprivation (BMSC+OGD/R group), overexpressed empty vector BMSC group cultured under glucose and oxygen deprivation(BMSC NC+OGD/R group), and overexpressed Nkx2.5 BMSC group cultured under glucose and oxygen deprivation (BMSC Nkx2.5+OGD/R group). The apoptosis rate of BMSCs in each group was detected via flow cytometry, and BMSC protein was extracted. The expression of caspase-3 and pro-caspase-3, caspase-8 and pro-caspase-8, caspase-9, and cytochrome C protein and expression of Nkx2.5 in the BMSCs of each group were detected by Western blot to determine the anti-apoptotic pathway in vitro. The model of myocardial infarction in mice was established by ligating the left anterior descending branch of coronary artery. The experiment was divided into five groups: sham surgery group, myocardial infarction untreated group, myocardial infarction tail vein injection of BMSC group, myocardial infarction tail vein injection of BMSC empty body group, myocardial infarction tail vein injection of BMSC overexpression Nkx2.5 group. The changes of cardiac function in mice were evaluated by echocardiography. Normal distribution econometric data were compared between groups using convenient analysis, and pairwise comparisons were conducted using LSD-t test. Results:The apoptosis rate of the BMSC+OGD/R group (12.98±1.24)% was higher than that of the BMSC group (7.82±0.42)%, and the difference was statistically significant ( P<0.001). The apoptosis rate of the BMSC NKx2.5+OGD/R group (11.26±0.22)% was lower than that of the BMSC+OGD/R group (12.98±1.24)% and the BMSC NC+OGD/R group (13.14±0.70)%, with statistically significant differences ( P<0.05). Compared to BMSC group ((0.36±0.08), (1.13±0.04), (0.36±0.06), (1.12±0.13), (1.23±0.08), (0.60±0.05), (0.67±0.14)), BMSC+OGD/R group ((1.05±0.10), (0.62±0.04), (1.07±0.09), (0.57±0.07), (0.55±0.08), (1.25±0.09), (0.71±0.04)) and BMSC NC+OGD/R group ((1.16±0.16), (0.64±0.06), (1.19±0.16), (0.56±0.06), (0.50±0.06), (1.28±0.06), (0.73±0.04)), the expression of Caspase-3 (0.72±0.08) and pro-caspase-3(0.89±0.09), Caspase-8 (0.63±0.08) and pro-caspase-8(0.85±0.12), Caspase-9 (0.87±0.09), cytochrome C (0.91±0.10), and Nkx2.5 (1.54±0.16) in BMSC Nkx2.5+OGD/R group was statistically significant (all P<0.05). In vivo experiments showed that the heart ejection fraction (29.05±7.07)% of mice treated with BMSC Nkx2.5 after myocardial infarction was significantly improved compared to the BMSC group (16.57±2.09)% and BMSC NC group (18.08±3.27)% (all P<0.05). Conclusion:BMSC Nkx2.5 may enhance the anti-apoptosis ability of BMSCs and improve cardiac function after myocardial infarction by inhibiting the death receptor pathway and the mitochondrial signal pathway .
