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| Effect of pVAX-eNOS transfection on the early endothelial progenitor cells from the patients with acute myocardial infarction |
| HUANG Xiaoyan1 WANG Sha2 SHEN Xin3 SUN Jingying1 DONG Xianghui4 JIN Zhankui4 SONG Baoguo2 LIANG Daoyan1 XU Cuixiang5 |
1.Department of Labratory Center, Shaanxi Provincial People′s Hospital, Shaanxi Province, Xi′an 710068, China;
2.Department of Cardial Surgery, Shaanxi Provincial People′s Hospital, Shaanxi Province, Xi′an 710068, China;
3.Department of Obstetric, Shaanxi Provincial People′s Hospital Maternity Hospital, Shaanxi Province, Xi′an 710068, China;
4.Department of Clinical Center, Shaanxi Provincial People′s Hospital, Shaanxi Province, Xi′an 710068, China;
5.Visitthe Cencer, Shaanxi Provincial People′s Hospital, Shaanxi Province, Xi′an 710068, China |
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Abstract Objective To investigate the feasibility of transfection of early endothelial progenitor cells (EPCs) in peripheral blood of patients with acute myocardial infarction (AMI) in vitro using pvax1-mediated nitric oxide synthase (eNOS) gene. Methods From December 2017 to June 2018, 30 cases of AMI patients admitted to Shaanxi Provincial People′s Hospital due to heart disease and diagnosed by coronary angiography were selected. The Ficoll density gradient centrifugation method was used to isolate mononuclear cells in the peripheral blood of patients with fasting veins and induce them to culture into EPCs for identification and purification of pvax-enos plasmids. EPCs were divided into three groups: eNOS transfection group, cationic polymer JetPEITM was used to transfect eNOS gene into EPCs in vitro. After transfection for 24 h, nitric oxide (NO) content in EPCs was detected by nitrate reductase method, and the ability of eNOS and vascular endothelial growth factor (VEGF) secretion by EPCs was detected by ELISA and Western blot. The proliferation of EPCs was detected by Wst-1 method, and the migration and adhesion of EPCs in each group were detected by Transwell chamber. Empty plasmid transfection group, only pVAX empty plasmid was transfected into EPCs. Control group, no transfection of EPCs. Results Pvax-enos gene was successfully transfected into early EPCs of AMI patients, and 24 h after transfection, the content of NO was significantly increased compared with the blank plasmid transfection group and the control group (P ＜ 0.01), The expression of eNOS was significantly increased (P ＜ 0.01), and the ability to secrete VEGF was significantly increased (P ＜ 0.01). The ability of proliferation, migration and adhesion of EPCs was significantly increased (P ＜ 0.01), and there was no significant difference between the empty plasmid transfection group and the control group (P ＞ 0.05). Conclusion Cationic polymer JetPEITM can successfully mediate the effective transfection of pvax-enos into early EPCs in AMI patients, and effectively enhance the proliferation, migration, adhesion and secretion of EPCs, which lays a foundation for the improvement of myocardial ischemia in AMI patients by autologous EPCs transplantation.
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