解剖学报

解剖学报 ›› 2020, Vol. 51 ›› Issue (3): 361-366.doi: 10.16098/j.issn.0529-1356.2020.03.008

• 细胞和分子生物学 • 上一篇    下一篇

京尼平对缺氧/复氧损伤后大鼠心肌细胞凋亡及自噬的影响

罗学秀 刘剑*   

  1. 重庆医科大学附属第一医院心血管内科，重庆 400016
  • 收稿日期:2019-10-22 修回日期:2020-01-06 出版日期:2020-06-06 发布日期:2020-06-06
  • 通讯作者: 刘剑 E-mail:liujian@hospital.cqmu.edu.cn
  • 基金资助:
    重庆市渝中区科技计划项目

Effect of genipin on cardiomyocyte apoptosis and autophagy in rat after hypoxia/reoxygenation injury#br#

LUO Xue-xiu LIU Jian*   

  1. Department of Cardiology, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
  • Received:2019-10-22 Revised:2020-01-06 Online:2020-06-06 Published:2020-06-06
  • Contact: LIU Jian E-mail:liujian@hospital.cqmu.edu.cn
  • Supported by:
    the Science and Technology Plan Project of Chongqing Yuzhong District

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摘要：

目的 探讨京尼平对缺氧/复氧(H/R)损伤后大鼠心肌细胞凋亡及自噬的影响。 方法 建立H/R损伤模型，体外培养的大鼠H9c2心肌细胞行缺氧12 h、复氧4 h。实验分为对照组（Con）、京尼平组(GE)、缺氧/复氧组(H/R)、缺氧/复氧+京尼平组(H/R+GE)。细胞计数试剂盒-8（CCK-8）检测细胞存活率，流式细胞仪检测细胞凋亡，透射电子显微镜观察自噬体，Western blotting检测Bax、Bcl-2、P62、Beclin1、LC3-Ⅱ、蛋白激酶B(Akt)、p-Akt、哺乳动物雷帕霉素靶蛋白（mTOR）和p-mTOR蛋白的表达。 结果 京尼平预处理增强了H/R损伤后的H9c2心肌细胞活力,抑制细胞凋亡及自噬体累积，降低自噬结构断面积与细胞质断面积的比值。Western blotting结果显示，京尼平预处理减少了H/R损伤后Bax、LC3-Ⅱ和Beclin1蛋白表达，增加Bcl-2、P62、p-Akt和p-mTOR蛋白表达。 结论 京尼平可以抑制H/R损伤后心肌细胞凋亡及自噬，其机制可能与上调Akt/mTOR信号通路有关。

关键词: 京尼平, 自噬, 缺氧/复氧, H9c2心肌细胞, 免疫印迹法, 大鼠

Abstract:

Objective  To investigate the effect of genipin on cardiomyocyte apoptosis and autophagy in rat after hypoxia/reoxygenation (H/R) injury.   Methods  The method  with hypoxia treatment of H9c2 cells for 12 hours and then reoxygenation treatment for 4 hours was used in the present study in order to establish H/R model. The H9c2 cells were divided into control group (Con), genipin group (GE), model group (H/R), model + genipin group (H/R+ GE). Cell viability was detected by cell counting kit-8(CCK-8). Cell apoptosis was determined by flow cytometry. Transmission electron microscopy was used to observe autophagosomes. The expression of Bax, Bcl-2, P62, Beclin1, LC3-Ⅱ, protein kinase B(Akt), p-Akt, mammalian target of rapamycin(mTOR), p-mTOR proteins were assessed by Western blotting.   Results Genipin pretreatment enhanced the cell viability, prevented cell apoptosis and autophagosome accumulation, and reduced the ratio of the cross-sectional areas of the autophagic structures to that of the cytoplasm after H/R injury in H9c2 cells. Western blotting showed that genipin pretreatment decreased the expression of Bax, LC3-Ⅱ, Beclin1 proteins and increased the expression of Bcl-2, p62, p-Akt, p-mTOR proteins after H/R injury.   Conclusion Genipin can inhibit H/R injury-induced apoptosis and autophagy, which may be through activating Akt/mTOR signaling pathway.

Key words: Genipin, Autophagy, Hypoxia/reoxygenation, H9c2 cardiomyocyte, Western blotting, Rat

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