上海交通大学学报(医学版)

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AT2R+与AT2R-人骨髓单个核细胞对于心肌梗死后心肌细胞的保护作用

杜明君,连锋,薛松   

  1. 上海交通大学 医学院附属仁济医院心血管外科, 上海 200127
  • 出版日期:2016-01-28 发布日期:2016-02-26
  • 通讯作者: 连锋, 电子信箱: dr.lianfeng@hotmail.com。
  • 作者简介: 杜明君(1989—), 男, 硕士生; 电子信箱: 919402904@qq.com。
  • 基金资助:

    上海市科委医学引导项目(124119a2100)

Protective effects of AT2R+ and AT2R- human bone marrow mononuclear cells on cardiomyocytes after incidence of myocardial infarction

DU Ming-jun,  LIAN Feng,  XUE Song   

  1. Department of Cardiovascular Surgery, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China
  • Online:2016-01-28 Published:2016-02-26
  • Supported by:

    Medicine Guidance Project of Science and Technology Commission of Shanghai Municipality, 124119a2100。

摘要:

目的 观察心肌梗死后人骨髓单个核细胞(BMMNCs)中AT2R的表达情况。比较AT2R+ BMMNCs与AT2R- BMMNCs在体外对受损心肌细胞的保护能力。方法 术中抽取患者骨髓。流式细胞仪分选出AT2R+ BMMNCs。利用8 μm孔径的Transwell迁移检测系统比较AT2R+ BMMNCs和AT2R- BMMNCs的迁移能力;分离SD乳鼠心肌细胞,并分别与AT2R+ BMMNCs和AT2R- BMMNCs共培养,RT-PCR检测AT2R+ BMMNCs和AT2R- BMMNCs向心肌细胞转分化的能力(心脏发育相关转录因子Nkx2.5和Gata4表达)的影响。在缺氧、无血清的条件下,AT2R+ BMMNCs和AT2R- BMMNCs分别与心肌细胞H9C2共培养,流式细胞术检测细胞凋亡。结果 流式细胞仪分选结果显示:心肌梗死患者骨髓样本中AT2R表达量较非心肌梗死患者上升约5.85%(P=0.039 8)。体外实验发现,AT2R+ BMMNCs在迁移和向心肌细胞转分化方面的能力均优于AT2R- BMMNCs;与AT2R- BMMNCs相比,AT2R+ BMMNCs对心肌细胞H9C2在缺氧和无血清条件下的抗凋亡作用更强。结论 心肌梗死后患者骨髓中AT2R的表达明显上调。AT2R+ BMMNCs可能对心肌梗死后的心肌细胞具有保护作用。

关键词: 骨髓单个核细胞, AT2受体, 迁移, 转分化, 心肌梗死

Abstract:

Objective To observe the expression of AT2R of human bone marrow mononuclear cells (BMMNCs) after the incidence of myocardial infarction (MI) and compare the ability of AT2R+ BMMNCs and AT2R- BMMNCs to protect impaired cardiomyocytes in vitro. Methods The bone marrow were drawn during operation and AT2R+ BMMNCs were isolated by flow cytometer. The migration ability of AT2R+ BMMNCs and AT2R-BMMNCs were compared by Transwell migration detection system with the pore size of 8 μm. The cardiomyocytes of neonatal SD mice were isolated and co-cultured with AT2R+ BMMNCs and AT2R- BMMNCs. The effects of AT2R+ BMMNCs and AT2R- BMMNCs on the transdifferentiation of cardiomyocytes (expressions of transcription factors Nkx2.5 and Gata4 relevant to the cardiac development) were detected by RT-PCR. AT2R+ BMMNCs and AT2R- BMMNCs were co-cultured with cardiomyocytes H9C2 under the conditions of hypoxia and non-serum and the cell apoptosis was detected by flow cytometry. Results Isolation results of flow cytometer showed that the expression of AT2R in bone marrow samples of patients with myocardial infarction was 5.85% higher than that of patients without myocardial infarction (P=0.039 8). In vitro experiment found that the migration ability and the ability of transdifferentiating towards cardiomyocytes of AT2R+ BMMNCs were better than those of AT2R- BMMNCs. Compared with AT2R- BMMNCs, the antiapoptosis effect of AT2R+ BMMNCs on cardiomyocytes H9C2 was stronger under the conditions of hypoxia and non-serum. Conclusion The expression of AT2R in bone marrow of patients with myocardial infarction significantly up-regulates. AT2R+ BMMNCs may protect the cardiomyocytes after the incidence of myocardial infarction.

Key words: bone marrow mononuclear cells, AT2 receptor, migration, transdifferentiation, myocardial infarction