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在人脐静脉内皮细胞中通过半定量逆转录聚合酶链反应评估能量平衡对人端粒酶逆转录酶(hTERT)可变剪接的影响。

Evaluation of Energy Balance on Human Telomerase Reverse Transcriptase (hTERT) Alternative Splicing by Semi-quantitative RT-PCR in Human Umbilical Vein Endothelial Cells.

作者信息

Behjati Mohaddeseh, Hashemi Mohammad, Kazemi Mohammad, Salehi Mansoor, Javanmard Shaghayegh Haghjooy

机构信息

Heart Failure Research Center, Cardiovascular Research Center, Cardiovascular Research Institute, Isfahan University of Medical Sciences, Isfahan, Iran.

Department of Cardiology, Isfahan University of Medical Sciences, Isfahan, Iran.

出版信息

Adv Biomed Res. 2017 Apr 17;6:43. doi: 10.4103/2277-9175.204591. eCollection 2017.

Abstract

BACKGROUND

Decreased high-energy phosphate level is involved in endothelial cell injury and dysfunction. Reduced telomerase activity in endothelial cells in parallel with reduced energy levels might be due to altered direction of alternative splicing machine as a complication of depleted energy during the process of atherosclerosis.

MATERIALS AND METHODS

Isolated human umbilical vein endothelial cells (HUVECs) were treated for 24 hours by oligomycine (OM) and 2-deoxy glucose (2-DG). After 24 hours, the effect of energy depletion on telomerase splicing pattern was evaluated using RT-PCR. Indeed, in both treated and untargeted cells, nitric oxide (NO) and von Willebrand factor (vWF) were measured.

RESULTS

ATP was depleted in treated cells by 43.9% compared with control group. We observed a slight decrease in NO levels ( = 0.09) and vWF ( = 0.395) in the setting of 49.36% ATP depletion. In both groups, no telomerase gene expression was seen. Telomerase and housekeeping gene expression were found in positive control group (colon cancer tissue) and sample tissue.

CONCLUSIONS

The absence of telomerase gene expression in HUVECs might be due to the mortality of these cells or the low level of telomerase gene expression in these cells under normal circumstances.

摘要

背景

高能磷酸水平降低与内皮细胞损伤和功能障碍有关。内皮细胞中端粒酶活性降低与能量水平降低同时出现,可能是由于在动脉粥样硬化过程中能量耗竭导致可变剪接机制方向改变所致。

材料与方法

将分离的人脐静脉内皮细胞(HUVECs)用寡霉素(OM)和2-脱氧葡萄糖(2-DG)处理24小时。24小时后,使用逆转录聚合酶链反应(RT-PCR)评估能量耗竭对端粒酶剪接模式的影响。实际上,在处理过的细胞和未处理的细胞中,均检测了一氧化氮(NO)和血管性血友病因子(vWF)。

结果

与对照组相比,处理过的细胞中三磷酸腺苷(ATP)耗竭了43.9%。在ATP耗竭49.36%的情况下,我们观察到NO水平(P=0.09)和vWF(P=0.395)略有下降。两组均未观察到端粒酶基因表达。在阳性对照组(结肠癌组织)和样本组织中发现了端粒酶和管家基因表达。

结论

HUVECs中未出现端粒酶基因表达可能是由于这些细胞的死亡或在正常情况下这些细胞中端粒酶基因表达水平较低。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8730/5414409/d5848af433cf/ABR-6-43-g001.jpg

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