中国组织工程研究

中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (20): 2949-2956.doi: 10.3969/j.issn.2095-4344.2016.20.009

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

前列腺素I2-内皮祖细胞保护氧化应激受损心肌细胞凋亡：
心肌电生理验证

张  靖，阮  璐，康  莉，崔晓海，张  佳   

  1. 西安交通大学第一附属医院胸外二科，陕西省西安市  710061
  • 收稿日期:2016-03-04 出版日期:2016-05-13 发布日期:2016-05-13
  • 通讯作者: 张佳，博士，主治医师，西安交通大学第一附属医院胸外二科，陕西省西安市 710061
  • 作者简介:张靖，男，1983年生，宁夏回族自治区固原市人，汉族，2009年西安交通大学毕业，硕士，主治医师，主要从事胸心外科临床与基础研究。
  • 基金资助:

    陕西省科学技术研究发展计划项目(2014KW23-02)；中央高校基本科研业务费专项资金资助

Cardiac protection of prostacyclin secreted from endothelial progenitor cells against oxidative stress-induced apoptosis: verified by cardiac electrophysiological tests

Zhang Jing, Ruan Lu, Kang Li, Cui Xiao-hai, Zhang Jia   

  1. Second Department of Thoracic Surgery, First Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710061, Shaanxi Province, China
  • Received:2016-03-04 Online:2016-05-13 Published:2016-05-13
  • Contact: Zhang Jia, M.D., Attending physician, Second Department of Thoracic Surgery, First Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710061, Shaanxi Province, China
  • About author:Zhang Jing, Master, Attending physician, Second Department of Thoracic Surgery, First Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710061, Shaanxi Province, China
  • Supported by:

    the Scientific and Technological Planning Project of Shaanxi Province, No. 2014KW23-02; the Fundamental Research Funds for the Central Universities

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摘要：

文章快速阅读：

文题释义：
内皮祖细胞：是血管内皮细胞的前体细胞，亦称为成血管细胞(angioblast)，在生理或病理因素刺激下，可从骨髓动员到外周血参与损伤血管的修复。1997年Asahara等首次证明循环外周血中存在能分化为血管内皮细胞的前体细胞，并将其命名为血管内皮祖细胞。内皮祖细胞在心脑血管疾病、外周血管疾病、肿瘤血管形成及创伤愈合等方面均发挥重要作用，并为缺血性疾病的研究治疗提供了新思路。
氧化应激：是指体内氧化与抗氧化作用失衡，倾向于氧化，导致中性粒细胞炎性浸润，蛋白酶分泌增加，产生大量氧化中间产物。氧化应激是由自由基在体内产生的一种负面作用，并被认为是导致衰老和疾病的一个重要因素。

摘要
背景：研究发现前列腺素I2及其类似物可预防压力超负荷性心肌肥厚，以及降低心肌缺血再灌注损伤，然而，其半衰期较短限制了其临床应用。作者前期以来源于骨髓的内皮祖细胞为载体构建了前列腺素I2-内皮祖细胞可持续产生前列腺素I2。
目的：观察前列腺素I2-内皮祖细胞对氧化应激引起的心肌细胞损伤的影响。
方法：体外培养大鼠心肌细胞H9c2，实验分为4组：①H2O2干预H9c2细胞4 h。其他3组H9c2在加入H2O2前1 h进行条件培养基预处理；②分别加入前列腺素I2-内皮祖细胞条件培养基；③内皮祖细胞的条件培养基；④加入PBS作为空白组。观察条件培养基-前列腺素I2-内皮祖细胞对内皮祖细胞管腔形成能力的影响；应用MTT和Hoechst 33342测定条件培养基对H2O2诱导的心肌细胞存活率及凋亡的影响。取雄性SD大鼠分离心肌，应用全细胞膜片钳技术检测条件培养基-前列腺素I2-内皮祖细胞对H2O2诱导的大鼠心肌细胞电生理活动的影响。
结果与结论：①条件培养基-前列腺素I2-内皮祖细胞组管腔形成能力明显强于条件培养基-内皮祖细胞组；②与条件培养基-内皮祖细胞组和空白对照组相比，条件培养基-前列腺素I2-内皮祖细胞预处理可显著降低H2O2诱导的心肌细胞凋亡和保存细胞存活(P < 0.01)；③条件培养基-前列腺素I2-内皮祖细胞预处理可预防H2O2诱导的早期后除极的发生，并且可缩短H2O2诱导的动作电位时程的延长(P < 0.01)；④结果提示前列腺素I2-内皮祖细胞通过分泌前列腺素I2保护氧化应激诱导损伤心肌细胞，可以心肌电生理活动作为保护效应的评价指标。

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；
ORCID: 0000-0001-8807-4685(张靖)组织工程

关键词: 组织构建, 内皮细胞, 祖细胞, 内皮祖细胞, 前列腺素I2, 凋亡, 心脏保护, 氧化应激

Abstract:

BACKGROUND: Prostacyclin (PGI2) and its analogs have been reported to prevent pressure overload-induced cardiac hypertrophy, and to reduce cardiac ischemia/reperfusion injury. However, clinical application of PGI2 is challenging due to its short half-life (< 2 minutes). Thus, we have generated PGI2 expressing rat endothelial progenitor cell strains (PGI2-EPCs) that constitutively secrete prostacyclin.
OBJECTIVE: To investigate the protective effect of PGI2-EPCs against oxidative stress-induced cardiomyocyte injury.
METHODS: Cultured H9c2 cells in vitro were assigned into four groups: H9c2 cells treated by H2O2 for 4 hours. H9c2 cells were pretreated by conditioned medium (collected form EPCs and PGI2-EPCs or collected form EPCs and PGI2-EPCs mixed with native EPCs) before the addition of H2O2. PBS instead of conditioned mediums served as negative control. The paracrine effect of PGI2-EPCs on in vitro angiogenesis of native EPCs was evaluated. MTT and Hoechst 33342 assays were used to examine the protective effect of conditioned medium on H2O2-induced rat embryonic cardiomyocyte apoptosis and cell viability. Finally, the effect of conditioned medium on the electric activities of adult cardiomyocytes was measured by whole-cell patch clamp techniques.
RESULTS AND CONCLUSION: When native EPCs mixed with conditioned medium of PGI2-EPCs, the total length of tubes was significantly longer compared with those mixed with CM of EPC. Rat embryonic cardiomyocytes pretreated with conditioned medium of PGI2-EPCs significantly reduced H2O2-induced apoptosis and preserved cell viability compared with pretreatment with EPC-conditioned medium and without pretreatment (P < 0.01). Pretreatment of rat adult cardiomyocytes with conditioned medium of PGI2-EPCs abolished H2O2-induced early after depolarization and shortened H2O2-induced action potential duration prolongation (P < 0.01) towards baseline. Our findings indicate that PGI2-EPCs protect against oxidative stress-induced cardiomyocyte injury through paracrine action. This Study provides the groundwork for an innovative cell therapy approach to treat ischemic heart disease.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words:  Prostaglandins I, Myocardium, Tissue Engineering

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