Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Apr 20;42(4):584-590.
doi: 10.12122/j.issn.1673-4254.2022.04.15.

[Role of PNPT1 in cardiomyocyte apoptosis induced by oxygen-glucose deprivation]

[Article in Chinese]
X Zhang  1   2 , X Wang  2 , Q Li  3 , Y Chen  2 , X Zhang  4 , P Wang  2 , M Yuan  5 , H Pei  1   2
Affiliations

[Role of PNPT1 in cardiomyocyte apoptosis induced by oxygen-glucose deprivation]

[Article in Chinese]
X Zhang et al. Nan Fang Yi Ke Da Xue Xue Bao. .

Abstract
in English, Chinese

Objective: To explore the effect of inhibiting polyribonucleotide nucleotidyl-transferase 1 (PNPT1) on oxygen-glucose deprivation (OGD)-induced apoptosis of mouse atrial myocytes.

Methods: Cultured mouse atrial myocytes (HL-1 cells) with or without OGD were transfected with PNPT1-siRNA or a negative control siRNA (NC-siRNA group), and the cell survival rate was detected using CCK-8 assay. The expression levels of ACTB and TUBA mRNA were detected with qPCR, and the protein expression of PNPT1 was detected with Western blotting. The apoptosis rate of the treated cells was determined with flow cytometry, the mitochondrial membrane potential was detected using JC-1 kit, and the mitochondrial morphology was observed using transmission electron microscope.

Results: With the extension of OGD time, the protein expression levels of PNPT1 increased progressively in the cytoplasm of HL-1 cells (P < 0.05). Transfection with PNPT1-siRNA significantly reduced PNPT1 expression in HL-1 cells (P < 0.05). Exposure to OGD significantly enhanced degradation of ACTB and TUBA mRNA (P < 0.05) and markedly increased the apoptosis rate of HL-1 cells (P < 0.05), and these changes were significantly inhibited by transfection with PNPT1-siRNA (P < 0.05), which obviously increased mitochondrial membrane potential and improved mitochondrial morphology of HL-1 cells exposed to OGD.

Conclusion: Inhibition of PNPT1 improves mitochondrial damage and reduces degradation of apoptotic-associated mRNAs to alleviate OGD-induced apoptosis of mouse atrial myocyte.

目的: 探讨多核糖核苷酸核苷转移酶1 (PNPT1)对氧糖剥夺（OGD）诱导心房肌细胞凋亡的影响和作用机制。

方法: 体外培养HL-1心房肌细胞，PNPT1-siRNA转染HL-1细胞。实验分组为：正常组（Control）、OGD组、NC-siRNA组（转染乱码RNA）、PNPT1- siRNA组、OGD+NC-siRNA组和OGD+PNPT1- siRNA组。通过CCK-8检测细胞存活率，qPCR检测ACTB mRNA和TUBA mRNA的含量，Western blot检测PNPT1蛋白水平，流式细胞术检测HL-1细胞凋亡率，JC-1试剂盒检测线粒体膜电位，透射电镜观察线粒体形态。

结果: 随OGD诱导时间延长，HL-1细胞质中PNPT1的蛋白表达水平呈上升趋势（P < 0.05）。与NCsiRNA相比，PNPT1-siRNA明显减少细胞内PNPT1表达。OGD条件下，ACTB mRNA和TUBA mRNA降解增加（P < 0.05），HL-1心房肌细胞凋亡率增加（P < 0.05）；相反地，PNPT1-siRNA则抑制ACTB mRNA和TUBA mRNA降解（P < 0.05），同时降低HL-1心房肌细胞凋亡率（P < 0.05），并且改善线粒体膜电位以及线粒体形态。

结论: 抑制PNPT1可改善线粒体损伤并减少凋亡相关mRNA的降解，从而减轻OGD诱导的HL-1心房肌细胞凋亡损伤。

Keywords: apoptosis-related mRNA degradation; myocardial cell apoptosis; oxygen-glucose deprivation; polyribonucleotide nucleotidyl-transferase 1.

PubMed Disclaimer

Figures

图 1
图 1
OGD诱导心肌细胞凋亡损伤 OGD induces apoptosis of cultured mouse cardiomyocytes (HL-1 cells). A: CCK-8 assay for detecting cell survival rate. B: Flow cytometry for detecting cell apoptosis rate. *P < 0.05 vs 0 h, #P < 0.05 vs control (n=6).
图 2
图 2
不同氧糖剥夺（OGD）时间处理后HL-1细胞PNPT1的蛋白表达 Protein expression of PNPT1 in HL-1 cells exposed to different OGD time. *P < 0.05 vs 0 h (n=6).
图 3
图 3
敲减PNPT1减轻OGD诱导的HL-1细胞凋亡 Knockdown of PNPT1 reduces OGD-induced apoptosis of HL-1 cells. A: PNPT1 protein expression detected by Western blotting. B: Flow cytometry for detecting cell apoptosis rate. *P < 0.05 (n=6).
图 4
图 4
敲减PNPT1抑制HL-1细胞OGD条件下凋亡相关mRNA的降解 Knockdown of PNPT1 inhibits degradation of apoptosis-related mRNA in HL-1 cells with OGD. A, B: qPCR for detecting the relative expression level of ACTB mRNA. C, D: qPCR for detecting relative expression level of TUBA mRNA. n=6. *P < 0.05 vs control, #P < 0.05 vs NC-siRNA, ＄P < 0.05 vs NC-siRNA+OGD (n=6).
图 5
图 5
敲减PNPT1减轻OGD条件下线粒体损伤 Knockdown of PNPT1 reduces mitochondrial damage in HL-1 cells exposed to OGD. A: Flow cytometry for detecting mitochondrial membrane potential (JC-1). B: Transmission electron microscopy for observing morphology of cell mitochondria. *P < 0.05 vs NC-siRNA+OGD (n=6).
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. 中国心血管健康与疾病报告编写组 中国心血管健康与疾病报告2020概要. 中国循环杂志. 2021;36(6):521–45. doi: 10.3969/j.issn.1000-3614.2021.06.001. - DOI
    1. Buja LM, Maximilian BL. Mitochondria in Ischemic Heart Disease. Adv Exp Med Biol. 2017;982:127–40. - PubMed
    1. Hu YF, Chen YJ, Lin YJ, et al. Inflammation and the pathogenesis of atrial fibrillation. Nat Rev Cardiol. 2015;12(4):230–43. doi: 10.1038/nrcardio.2015.2. - DOI - PubMed
    1. Jame S, Barnes G. Stroke and thromboembolism prevention in atrial fibrillation. Heart. 2020;106(1):10–7. doi: 10.1136/heartjnl-2019-314898. - DOI - PMC - PubMed
    1. Tsai CF, Yang SF, Lo CH, et al. Role of the ROS-JNK signaling pathway in hypoxia-induced atrial fibrotic responses in HL-1 cardiomyocytes. Int J Mol Sci. 2021;22(6):3249–57. doi: 10.3390/ijms22063249. - DOI - PMC - PubMed

LinkOut - more resources