Effect of Carabin overexpression on oxygenglucose deprivation-induced apoptosis in cultured human ventricular myocytes

Qing WU; Haoqi YAO; Kuoju CHENG; Jianfang TANG; He HUANG; Jian CHEN; Hong LI; Tiande YANG

doi:10.16016/j.1000-5404.201807146

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Effect of Carabin overexpression on oxygenglucose deprivation-induced apoptosis in cultured human ventricular myocytes

Basic Medicine | 更新时间：2026-01-09

- Effect of Carabin overexpression on oxygenglucose deprivation-induced apoptosis in cultured human ventricular myocytes
- Journal of Third Military Medical University Vol. 40, Issue 22, Pages: 2068-2073(2018)
- 作者机构：
  
  1.400037 重庆，陆军军医大学(第三军医大学)第二附属医院麻醉科
  2.400038 重庆，陆军军医大学(第三军医大学)高原军事医学系
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81370210)
- DOI：10.16016/j.1000-5404.201807146
  CLC： R322.11;R394-33;R394.2
- Received：28 July 2018，
  
  Revised：2018-09-28，
  
  Published：30 November 2018
- 稿件说明：
移动端阅览
Qing WU, Haoqi YAO, Kuoju CHENG, et al. Effect of Carabin overexpression on oxygenglucose deprivation-induced apoptosis in cultured human ventricular myocytes[J]. Journal of Third Military Medical University, 2018, 40(22): 2068-2073.
DOI：

Qing WU, Haoqi YAO, Kuoju CHENG, et al. Effect of Carabin overexpression on oxygenglucose deprivation-induced apoptosis in cultured human ventricular myocytes[J]. Journal of Third Military Medical University, 2018, 40(22): 2068-2073. DOI： 10.16016/j.1000-5404.201807146.

摘要

目的

探讨Carabin基因过表达对氧糖剥夺(oxygen glucose deprivation

OGD)诱导的人心室肌细胞株AC16凋亡的作用。

方法

构建Carabin过表达及空载稳定细胞株，将AC16细胞分为空载常氧组、Carabin过表达常氧组、空载OGD 24 h组和Carabin过表达OGD 24 h组。空载常氧组及Carabin过表达常氧组置于正常条件下培养；空载OGD 24 h组和Carabin过表达OGD 24 h组置于Hanks液中放入缺氧恒温细胞培养箱培养24 h(1%O

-5%CO

-94%N

)。采用倒置显微镜观察细胞形态变化；流式细胞术检测心肌细胞凋亡率及存活率；TUNEL染色检测细胞凋亡情况；通过酶标仪检测心肌细胞凋亡蛋白酶Caspase-3和心肌细胞损伤标志物LDH的活性；Western blot检测细胞凋亡蛋白Caspase-3的表达水平。

结果

AC16细胞OGD 24 h后，细胞内Carabin的mRNA及蛋白水平显著降低。与空载常氧组比较，空载OGD 24 h组及Carabin过表达OGD 24 h组的细胞凋亡率、Caspase-3活性、LDH活性及Caspase-3蛋白表达水平均显著升高(

0.05)。与空载OGD 24 h组比较，Carabin过表达OGD 24 h组的细胞凋亡率、Caspase-3活性、LDH活性及Caspase-3蛋白表达水平显著降低(

0.05)。

结论

Carabin过表达能显著减轻OGD诱导后AC16细胞的凋亡。

Abstract

Objective

To investigate the effect of Carabin overexpression on apoptosis of human ventricular myocytes induced by oxygenglucose deprivation (OGD)

in vitro

Methods

Human ventricular myocyte AC16 cell line was transfected with a lentiviral vector carrying Carabin gene to induce Carabin overexpression or with a negative control vector. The transfected cells were cultured for 24 h under routine conditions or under OGD condition

s (in Hank's solution in a hypoxia incubator with 1% O

5% CO

and 94% N

). The changes in the morphology of AC16 cells were observed under an inverted microscope

and flow cytometry was used to determine the apoptosis rate and survival rate of the cells. The apoptotic cells were detected with TUNEL staining

and the activity of Caspase-3 and LDH in the cells were evaluated with enzyme labeling. Western blotting was performed to determine the protein expression of Caspase-3 in the cells in different culture conditions.

Results

OGD for 24 h significantly lowered the mRNA and protein levels of Carabin in AC16 cells. In the cells transfected with Carabin-overexpressing vector and the negative control vector

OGD for 24 h resulted in significantly increased cell apoptosis rate

Caspase-3 activity

LDH activity

and Caspase-3 protein expression as compared with the routinely cultured cells transfected with the negative control vector (

0.05). Compared with the negative control cells

the cells overexpressing Carabin were more resistant to OGD and showed significantly lower apoptotic rate

Caspase-3 and LDH activities

and Caspase-3 protein expression after OGD for 24 h (

0.05).

Conclusion

The overexpression of Carabin can significantly reduce the apoptosis of AC16 cells induced by OGD.

关键词

Keywords

references

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