低温缺血-再灌注对离体大鼠心房肌Kir2.1和CaMKⅡ表达的影响

doi:10.3969/j.issn.1006-5725.2023.21.010

摘要/Abstract

摘要：

目的探讨再灌注房性心律失常心房肌复极时程延长的分子机制。方法采用随机数字表法将16只由雄性SD大鼠制备的Langendorff离体心脏灌注模型分为对照组（C组，n = 8）和低温缺血-再灌注组（IR组，n = 8）。根据再灌注后是否发生房性心律失常，将IR组进一步细分为再灌注非房性心律失常亚组（N-RAA组）和再灌注房性心律失常亚组（R-AA组）。C组使用37 ℃ K-H液平衡灌注120 min。IR组使用37 ℃ K-H液平衡灌注30 min后停止，注射4 ℃ Thomas液（20 mL/kg）使心脏停跳60 min，停跳30 min时使用半量4 ℃ Thomas液（10 mL/kg）对离体心脏进行复灌［停跳期间用低温Thomas液（4 ℃）对心脏进行保护］，后再次灌注37 ℃ K-H液30 min。记录平衡灌注30 min（T₀）、平衡灌注105 min/再灌注15 min（T₁）和平衡灌注120 min/再灌注30 min（T₂）时右心房单相动作电位（MAP），测量单相动作电位复极50%和90%的时程（MAPD₅₀和MAPD₉₀）。电生理指标监测完后用Western blot检测右心房组织内向整流钾通道2.1（Kir2.1）和Ca²⁺/CaM依赖激酶Ⅱ（CaMKⅡ）的表达。结果与T₀时点比较，R-AA组T₁、T₂时MAPD₅₀、MAPD₉₀明显延长（P < 0.05）；与C组比较，R-NAA组和R-AA组T₁、T₂时MAPD₉₀明显延长（P < 0.05）；与R-NAA组比较，R-AA组T₁、T₂时MAPD₅₀、MAPD₉₀明显延长（P < 0.05）。Western blot 结果显示，R-NAA组和R-AA组Kir2.1表达明显少于C组（P < 0.05），且R-AA组明显少于R-NAA组（P < 0.05）；R-NAA组和R-AA组CaMKⅡ表达较C组明显增加（P < 0.05），且R-AA组 CaMKⅡ表达较R-NAA组明显增加（P < 0.05）。结论低温缺血-再灌注房性心律失常大鼠心房肌复极时程延长可能与Kir2.1表达下调和CaMKⅡ 表达增加有关。

关键词: 缺血-再灌注, 心房肌, 复极时程, 内向整流钾通道2.1, Ca²⁺/CaM依赖激酶Ⅱ

Abstract:

Objective To explore the molecular mechanism of prolonged atrial repolarization in rats with reperfusion atrial arrhythmia. Methods Sixteen Langendorff isolated heart perfusion models made by male SD rats were randomly divided into control group （group C，n = 8） and hypothermic ischemia-reperfusion group （group IR，n = 8）. According to the occurrence of atrial arrhythmia after reperfusion，group IR was further subdivided into reperfusion non-atrial arrhythmia subgroup （group N-RAA） and reperfusion atrial arrhythmia subgroup （group R-AA ）.Group C was perfused with 37 ℃ K-H solution for 120 min. In group IR， the isolated heart was perfused with 37 ℃ K-H solution for 30 min and stopped， and the isolated heart was perfused with 4 ℃ Thomas solution （20 mL/kg） for 60 mins. When the heart stopped for 30 mins， the isolated heart was perfused with a half dose of 4 ℃ Thomas solution （10 ℃）. During cardioplegia， the isolated heart was protected by low temperature Thomas solution （4 ℃）， and then reperfused for 30 mins with 37 ℃ K-H solution. The monophasic action potential （MAP） of the right atrium was recorded at balanced perfusion for 30 mins （T₀），balanced perfusion for 105 mins in group C/reperfusion for 15 mins in group IR（T₁） and balanced perfusion for 120 mins in group C/reperfusion for 30 min in group IR（T₂）； The duration of 50% and 90% repolarization of monophasic action potential （MAPD₅₀ and MAPD₉₀） was measured. After electrophysiological monitoring， the expression of Kir2.1 and CaMKⅡ in right atrium was detected by Western blot. Results Compared with T₀， MAPD₅₀ and MAPD₉₀ at T₁ and T₂ were significantly prolonged in group R-AA （P < 0.05）， and MAPD₉₀ at T₁ and T₂ in group R-NAA and group R-AA were significantly longer than those in group C （P < 0.05）.Compared with group R-NAA， MAPD₅₀ and MAPD₉₀ in group R-AA were significantly prolonged at T₁ and T₂ （P < 0.05）.The results of Western blot showed that the expression of Kir2.1 in group R-NAA and group R-AA was significantly lower than that in group C （P < 0.05）， and that in group R-AA was significantly lower than that in group R-NAA （P < 0.05）.The expression of CaMKⅡ in group R-NAA and group R-AA was significantly higher than that in group C （P < 0.05）， and the expression of CaMKⅡ in group R-AA was significantly higher than that in group R-NAA. Conclusion The prolonged duration of atrial repolarization in rats with hypothermic ischemia-reperfusion atrial arrhythmia may be related to the down-regulation of Kir2.1 expression and the up-regulation of CaMKⅡ expression.

Key words: ischemia-reperfusion, atrial myocardium, duration of repolarization, inwardly rectifying K channels 2.1, Ca²⁺-calmodulin kinase Ⅱ

中图分类号:

何幼芹,高鸿,种朋贵,刘艳秋,佟睿,吴学艳. 低温缺血-再灌注对离体大鼠心房肌Kir2.1和CaMKⅡ表达的影响[J]. 实用医学杂志, 2023, 39(21): 2750-2753.

Youqin HE,Hong GAO,Penggui CHONG,Yanqiu LIU,Rui TONG,Xueyan. WU. Effect of hypothermic ischemia⁃reperfusion on the expression of Kir2.1 and CaMKⅡ in isolated rat atrial myocardium[J]. The Journal of Practical Medicine, 2023, 39(21): 2750-2753.

图/表 2

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项目	组别	n	T₀	T₁	T₂
MAPD₅₀	C组	8	19.15 ± 2.03	19.42 ± 1.99	20.10 ± 2.23
	R-NAA组	5	19.21 ± 2.17	23.95 ± 2.10^*▲	21.69 ± 2.71
	R-AA组	3	19.09 ± 2.51	29.05 ± 3.81^*△▲	28.22 ± 3.37^*△▲
MAPD₉₀	C组	8	42.26 ± 1.62	43.82 ± 2.85	43.14 ± 3.74
	R-NAA组	5	43.34 ± 2.46	48.76 ± 2.66^*▲	47.58 ± 4.56^*▲
	R-AA组	3	43.48 ± 3.81	54.22 ± 3.85^*△▲	52.15 ± 4.99^*△▲

组别	n	Kir2.1	CaMKⅡ
C组	8	1.99 ± 0.44	0.49 ± 0.15
R-NAA组	5	1.30 ± 0.39^*	1.14 ± 0.25^*
R-AA组	3	0.46 ± 0.23^*△	1.65 ± 0.30^*△

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