Effect of ethanol and its metabolites on acetylcholine-sensitive K+ channel Kir3.1 protein expression of neonatal rat primary atrial cardiomyocytes
10.3760/cma.j.issn.0253-3758.2015.07.009
- VernacularTitle:乙醇及其代谢产物对新生大鼠原代培养心房肌细胞乙酰胆碱敏感型钾通道Kir3.1蛋白表达的影响
- Author:
Yuanyuan ZHAO
1
;
Jinghan SUN
;
Jun HU
;
Ni BO
;
Bo YU
Author Information
1. 中国医科大学附属第一医院心血管内科
- Keywords:
Ethanol;
Acetaldehyde;
Myocytes,cardiac;
G Protein-coupled inwardly-rectifying potassium channels;
Arrhythmias
- From:
Chinese Journal of Cardiology
2015;43(7):609-613
- CountryChina
- Language:Chinese
-
Abstract:
Objective To identify the effect of ethanol and its metabolite acetaldehyde on acetylcholine-sensitive K + channel Kir3.1 protein expression,and explore the potential role of this channeland acetaldehyde in arrhythmia caused by acute alcoholic intoxication.Methods Primary atrial cardiomyocytes were isolated from 150 newborn SD rats by typsin and type Ⅱ collagenase,cultured and troponin Ⅰ was determined by immunofluorescence.Cell survival in 200-800 mmol/L ethanol or 50-500 μmol/L acetaldehyde treated cells for 24 hours was measured by CCK-8 assay to determine the concentration of ethanol and acetaldehyde for inducing apoptosis in cardiomyocytes.The highest non-apoptotic concentration (200 mmol/L) of ethanol and acetaldehyde(100 μmol/L) was used in the main study.Kir3.1 protein expression was detected by Western blot.Results (1) Cellular immunofluorescence results showed that cultured cells are cardiomyocytes,and more than 90％ of these cells are troponin Ⅰ positive.(2)CCK-8 assay demonstrated that the survival rate of cardiomyocytes in the groups treated by ethanol over 400 mmol/L for 24 hours or acetaldehyde over 400 μmoL/L was significantly lower than that of the control group (P ＜ 0.05),while the survival rate was similar in cardiomyocytes treated by ethanol less than 200 mmol/L or acetaldehyde less than 350 μmol/L for 24 hours and the control group (P ＞ 0.05).(3) Western-bolt assay revealed that ethanol and acetaldehyde treatment for 24 hours upregulated Kir3.1 protein expression in primary atrial cardiomyocytes of newborn SD rats by (44.52 ± 23.07) ％ and (45.04 ± 22.01) ％ respectively compared with the control group(all P ＜ 0.01).Conclusions Acute ethanol and acetaldehyde treatment could significantly upregulate the protein expression of acetylcholine-sensitive K + channel Kir3.1,this might serve as a potential mechanism for arrhythmia caused by acute alcoholic intoxication.
