Effects of ischemia-reperfusion on endothelial progenitor cell function and iNOS, eNOS expression / 中国医师杂志

ABSTRACT

Objective To explore the effects of ischemia and ischemia reperfusion on the proliferation,apoptosis,migration ability,inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) expression of endothelial progenitor cells (EPCs).Methods Collection of peripheral blood from volunteers and culture of endothelial progenitor cells in vitro.The cells were divided into three groupscontrol group,hypoxia group and hypoxia reoxygenation group.Methyl thiazolyl tetrazolium (MTT) assay was used to detect cell proliferation.Transwell chamber method was used to detect cell migration.Cell apoptosis was detected by flow cytometry.Western blot was used to detect iNOS and eNOS expressions.Results A confocal microscope was used to observe the basic adherence of the cells to the wall for about 3 days,and the area became larger.After 7 d of single nucleus cell culture,the growth of colony-like pattern was more than that of spindle.The cell counts of the three groups in the microscope were (1.83 ± 0.92),(5.07± 0.84),(2.11 ± 0.74).Compared with the control group (0.24 ± 0.04),the hypoxia group (0.62± 0.06) could promote EPCs proliferation,and the difference was statistically significant (t =12.142,P ＜ 0.05);While there was no significant difference between the hypoxia reoxygenation group (0.39 ± 0.06) and the control group (P ＞ 0.05).The number of cell migration in the hypoxia group (18.28 ± 2.05) and hypoxic complex oxygen group (14.08 ± 2.11) was not statistically significant compared with the control group (15.14 ± 1.25) (P ＞ 0.05).The apoptosis rate in hypoxia group (34.57 ±0.42)％ and hypoxia reoxygenation group (41.08 ± 0.44)％ was significantly higher than that in control group (24.83 ± 0.38) ％ (x2 =13.427,15.084,P ＜ 0.05).The apoptosis rate of hypoxia reoxygenation group was significantly higher than that of hypoxia group (x2 =9.657,P ＜ 0.05).The expression of iNOS in hypoxia group and hypoxia reoxygenation group was significantly higher than that in control group,and the difference was statistically significant (P ＜ 0.05).Conclusions Ischemia could promote the proliferation of EPCs,and increase the expression of iNOS,but the expression of EPCs was down-regulated after reperfusion.