白细胞介素37过表达促进急性心肌梗死患者外周血内皮祖细胞的增殖和迁移

Interleukin-37 overexpression promotes proliferation and migration of peripheral blood endothelial progenitor cells in patients with acute myocardial infarction

目的:探究白细胞介素(IL)-37对急性心肌梗死患者外周血内皮祖细胞(EPCs)增殖和迁移的影响和机制。方法:从6例急性心肌梗死患者和6例年龄性别差异无统计学意义的同期进行体检的健康志愿者外周血中分离并培养EPCs。检测IL-37 mRNA和蛋白表达水平。将急性心肌梗死患者EPCs随机分成3组:对照组、pcDNA3.1组和IL-37组。其中对照组不进行任何处理,pcDNA3.1组和IL-37组分别使用Lipofectamine3000试剂转染pcDNA3.1质粒和pcDNA3.1-IL-37质粒。CCK-8检测细胞活力,Transwell检测迁移细胞数目,qRT-PCR检测IL-37和基质金属蛋白酶(MMP)9 mRNA表达水平,Western blot检测IL-37、增殖细胞核抗原(PCNA)、MMP9、磷酸化-Akt(p-Akt)、Akt、p-mTOR、mTOR、p-p70S6K、p70S6K、p-ERK1/2和ERK1/2蛋白水平表达。结果:与健康志愿者相比,IL-37在急性心肌梗死患者外周血EPCs中表达上调(P<0.01)。与对照组相比,IL-37组中细胞活力、迁移细胞数目和IL-37、PCNA、MMP9表达以及p-Akt/Akt、p-mTOR/mTOR、p-p70S6K/p70S6K均增加,差异具有统计学意义(P<0.05);但pcDNA3.1组中以上指标差异均无统计学意义(P>0.05)。与对照组相比,p-ERK1/2/ERK1/2在IL-37组和pcDNA3.1组中差异均无统计学意义(P>0.05)。结论:IL-37过表达促进急性心肌梗死患者外周血EPCs的增殖和迁移,其可能通过激活Akt/mTOR通路发挥作用,与ERK1/2通路无关。

Objective:To investigate effect and mechanism of interleukin(IL)-37 on proliferation and migration of peripheral blood endothelial progenitor cells(EPCs)in patients with acute myocardial infarction.Methods:EPCs were isolated and cultured from peripheral blood of six patients with acute myocardial infarction and six age and sex-matched healthy volunteers who take a physical examination at the same time. mRNA and protein levels of IL-37 were detected. EPCs of patients with acute myocardial infarction were randomly divided into three groups:control group,pcDNA3.1 group and IL-37 group. Cells in control group did not receive any treatment,and cells in pcDNA3.1 group and IL-37 group were transfected with pcDNA3.1 and pcDNA3.1-IL-37 plasmids,respectively,using Lipofectamine 3000 reagent. Cell viability was detected by CCK-8,number of migrated cells was analyzed by Transwell assay.qRT-PCR was used to determine mRNA levels of IL-37 and matrix metalloproteinase(MMP)9. Western blot was used to measure protein levels of IL-37,proliferating cell nuclear antigen(PCNA),MMP9,phosphorylated-Akt(p-Akt),Akt,p-mTOR,mTOR,p-p70S6K,p70S6K,p-ERK1/2 and ERK1/2.Results:IL-37 expression was higher in peripheral blood EPCs of patients with acute myocardial infarction than that in healthy controls(P<0.01). Compared with control group,cell viability,number of migrated cells,expressions of IL-37,PCNA and MMP9,ratio of p-Akt/Akt,p-mTOR/mTOR and p-p70S6K/p70S6K were significantly enhanced in IL-37 group(P<0.05);whereas these indexes were not changed in pcDNA3.1 group(P>0.05). Compared with control group,p-ERK1/2/ERK1/2 was not altered in IL-37 group and pcDNA3.1 group(P>0.05).Conclusion:Overexpression of IL-37 promoted proliferation and migration of EPCs in peripheral blood of patients with acute myocardial infarction,which might play a role through activation of Akt/mTOR pathway,but not ERK1/2 pathway.