中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (在线): 1-7.doi: 10.3969/j.issn.2095-4344.2016.00.000

• 心脏组织构建 cardiac tissue construction •    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
  • 出版日期:2019-03-28 发布日期:2019-03-28
  • 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 

  • Online:2019-03-28 Published:2019-03-28
  • Contact: 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
  • 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：

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：① H2O2干预H9c2细胞4 h。其他3组H9c2在加入H2O2前1h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。

关键词: 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, we measured the effect of conditioned medium on the electric activities of adult cardiomyocytes 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 afterdepolarization and shortened H2O2-induced action potential duration prolongation (P < 0.01) towards baseline. PGI2-EPCs protect against oxidative stress-induced cardiomyocyte injury through paracrine action. Our findings provide the groundwork for an innovative cell therapy approach to treat ischemic heart disease.