Inhibition of carotid restenosis by Krüppel like factor 4 gene-modified bone marrow mesenchymal stem cells
Objective To construct a recombinant lentiviral vector carrying the Krüppel like factor 4(KLF4)gene,establish bone marrow mesenchymal stem cells(BMSCs)overexpressing the KLF4 gene,and observe the changes in vascular proliferation after carotid artery injury.Methods The lentivirus was used as a vector to mediate KLF4 gene transfection into BMSCs.Totally,24 10-week-old SD rats were ran-domly divided into three groups:sham surgery group,model group,and KLF4 group(injected with KLF4-modified BMSCs).An animal model of carotid artery endothelial injury in rats was constructed using a bal-loon catheter.After 14 days,the animals were euthanized by femoral artery bleeding,and hematoxylin and eosin(HE)staining was used to observe the changes in the thickness and morphology of the common carot-id artery vessel walls in each group.The Griess method was used to detect the content of nitric oxide(NO)in the serum of experimental animals in each group.Western blotting was used to detect the expression lev-els of endothelial nitric oxide synthase(eNOS)and KLF4 proteins in rat carotid arteries.One-way analysis of variance was used for comparison between groups.Results The endothelial cell hyperplasia in the mod-el group was severer than that in the sham operation group(1.68±0.29 vs.0.01±0.01,F=244.345,P＜0.05),and the difference was statistically significant,while the endothelial cell hyperplasia in the KLF4 group was milder than that in the model group(0.14±0.01 vs.1.68±0.29,F=244.345,P＜0.05).The serum NO concentration in the model group was significantly lower than that in the sham group(7.22±1.40 vs.21.29±1.83,F=171.000,P＜0.05),and that in the KLF4 group was significantly higher than that in the model group(16.34±1.33 vs.7.22±1.40,F=171.000,P＜0.05).Western blotting showed that the ratios of KLF4/β-actin in the sham operation group,model group and KLF4 group were 0.564±0.015,0.626±0.023 and 0.916±0.042,respectively,and the differences were statistical-ly significant(F=125.121,P＜0.05),and the ratios of eNOS/β-actin in the sham operation group,model group and KLF4 group were 0.852±0.043,0.383±0.017,0.707±0.014 respectively,and the difference was statistically significant(F=223.879,P＜0.05).Conclusion KLF4 can positively regu-late the expression of eNOS,thereby increasing the concentration of NO in blood vessels to inhibit the mi-gration and proliferation of endothelial cells and suppress vascular stenosis caused by carotid artery injury.
Krüppel like factor 4Gene therapySlow virusEndothelial nitric oxide syn-thaseCarotid artery
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