切换至 "中华医学电子期刊资源库"

中华临床医师杂志(电子版) ›› 2019, Vol. 13 ›› Issue (11) : 855 -859. doi: 10.3877/cma.j.issn.1674-0785.2019.11.012

所属专题: 文献

基础研究

地尔硫䓬对离体大鼠心肌缺血再灌注损伤影响的研究
张云盛1, 滕天明2, 张文娟2,()   
  1. 1. 300308 天津医科大学总医院空港医院心血管内科
    2. 300052 天津医科大学总医院心血管内科
  • 收稿日期:2019-03-17 出版日期:2019-06-01
  • 通信作者: 张文娟
  • 基金资助:
    天津市高等科技发展基金计划项目(20110151); 天津市科技计划项(15YFYZSY00020)

Effect of diltiazem on myocardial ischemia-reperfusion injury in isolated rat hearts

Yunsheng Zhang1, Tianming Teng2, Wenjuan Zhang2,()   

  1. 1. Department of Cardiology, Tianjin Medical University General Hospital Airport Hospital, Tianjin 300308, China
    2. Department of Cardiology, Tianjin Medical University General Hospital, Tianjin 300052, China
  • Received:2019-03-17 Published:2019-06-01
  • Corresponding author: Wenjuan Zhang
  • About author:
    Corresponding author: Zhang Wenjuan, Email:
引用本文:

张云盛, 滕天明, 张文娟. 地尔硫䓬对离体大鼠心肌缺血再灌注损伤影响的研究[J/OL]. 中华临床医师杂志(电子版), 2019, 13(11): 855-859.

Yunsheng Zhang, Tianming Teng, Wenjuan Zhang. Effect of diltiazem on myocardial ischemia-reperfusion injury in isolated rat hearts[J/OL]. Chinese Journal of Clinicians(Electronic Edition), 2019, 13(11): 855-859.

目的

探讨地尔硫䓬对离体大鼠心肌缺血再灌注损伤的影响及其机制。

方法

33只大鼠随机分为3组:正常对照组(N组)、缺血再灌注组(I-R组)、地尔硫䓬+缺血再灌注组(D/I-R组)。N组持续灌流K-H液150 min;I-R组K-H液稳定灌流30 min后,结扎前降支30 min,继以K-H液再灌注90 min;D/I-R组给予地尔硫䓬(5 μmo/L)再灌注15 min,继以K-H液再灌流75 min。记录并分析各组大鼠左心室发展压、左心室内压力最大上升/下降速率(±dp/dtmax)、再灌注心律失常评分、心肌梗死面积以及各组左心室心尖组织的线粒体乙醛脱氢酶2(ALDH2)、Bcl-2以及Bax的mRNA基因与蛋白表达水平。

结果

(1)左心室发展压D/I-R组再灌注30 min与45 min比I-R组压力明显升高[(92.68±5.09)mmHg vs(75.77±5.33)mmHg;(90.39±4.29)mmHg vs(72.34±7.49)mmHg;1 mmHg=0.133 kPa],差异均具有统计学意义(F=72.81、51.92,P均=0.001)。(2)±dp/dtmax D/I-R组再灌注30 min与45 min时均比I-R组升高[+dp/dtmax:(2885.45±286.47)mmHg vs (2063.64±105.57)mmHg;(2712.73±236.52)mmHg vs(2053.64±92.33)mmHg;-dp/dtmax:(2214.55±104.63)mmHg vs (1710.91±217.97)mmHg;(2119.09±84.43)mmHg vs(1544.55±207.72)mmHg],差异均具有统计学意义(F=64.22、70.55、69.77、54.64,P均=0.001)。(3)N组仅有室性期前收缩的发生,未发生心室颤动、室性心动过速;D/I-R组出现室性期前收缩的个数较N组增加,差异具有统计学意义(P=0.001);I-R组室性心动过速发生率高于D/I-R组,心室颤动时程与室性心动过速时程均较D/I-R组增加,差异具有统计学意义(P=0.013、0.049、0.001);再灌注心律失常评分I-R组评分[5(3,6),57.36]高于D/I-R组[3(1,4),34.77],差异具有统计学意义(P=0.001)。(4)心肌梗死面积I-R组高于D/I-R组[(55.51±1.43)% vs (17.01±1.13)%],差异具有统计学意义(P<0.01)。(5)I-R组线粒体ALDH2表达明显减少。D/I-R组线粒体ALDH2表达与I-R组比较减少,但差异无统计学意义(P=0.11),Bcl-2、Bax的表达均增加,D/I-R组Bcl-2/Bax的比值较I-R组增大(0.44 vs 0.22),差异具有统计学意义(P=0.001)。

结论

地尔硫䓬处理后能够减少缺血再灌注损伤。其保护作用机制之一可能是通过上调Bcl-2下调Bax的基因和蛋白表达,减少心肌细胞的凋亡,但其并不是通过上调线粒体ALDH2的基因与蛋白来实现。

Objective

To investigate the effects of diltiazem on myocardial ischemia-reperfusion injury (MIRI) and the underlying mechanisms.

Methods

Wistar rats were randomly divided into three groups: normal group (N group), ischemia-reperfusion group (I-R group), diltiazem+ ischemia-reperfusion group (D/I-R group). The isolated rat hearts in different groups were given different perfusion treatments: The N group was given continuous perfusion of K-H liquid for 150 min; the I-R group was given stable perfusion of K-H liquid for 30 min followed by ligating the left anterior descending coronary artery for 30 min, and K-H liquid reperfusion for 90 min; the D/I-R group underwent reperfusion with diltiazem (5 μmo/L) for 15 min and reperfusion with K-H liquid for 75 min. Left ventricular cardiac function (LVDP), maximal rise/fall rate of left ventricular pressure (±dp/dtmax), reperfusion arrhythmia (RA) score, calculated myocardial infarct (MI) size, and ALDH2, Bcl-2, and BAX expression in the left ventricular apex were recorded in each group.

Results

The LVDP at 30 min and 45 min in the D/I-R group was significantly higher than that of the I-R group, respectively [(92.68±5.09) mmHg vs (75.77±5.33) mmHg, F=72.81, P=0.001; (90.39±4.29) mmHg vs (72.34±7.49) mmHg, F=51.92, P=0.001]. The ±dp/dtmax at 30 min and 45 min in the D/I-R group were significantly higher than that of the I-R group, respectively [+ dp/dtmax: (2885.45±286.47) mmHg vs (2063.64±105.57) mmHg, F=64.22, P=0.001 and (2712.73±236.52) mmHg vs (2053.64±92.33) mmHg, F=70.55, P=0.001; -dp/dtmax: (2214.55±104.63) mmHg vs (1710.91±217.97) mmHg, F=69.77, P=0.001 and (2119.09±84.43) mmHg vs (1544.55±207.72) mmHg, F=54.64, P=0.001, respectively]. The number of ventricular pre-contractions in the I-R group was significantly higher than that of the D/I-R group (P=0.001). The incidence of ventricular tachycardia, the time history of ventricular fibrillation, and the duration of ventricular tachycardia in the I-R group were also significantly higher than those of the D/I-R group (P=0.013, 0.049, and 0.001, respectively). The reperfusion arrhythmia score in the I-R group [5(3, 6), 57.36] was significantly higher than that of the D/I-R group [3(1, 4), 34.77] (P=0.001). Compared with the I-R group, the D/IR group had significantly smaller MI size [(55.51±1.43)% vs (17.01±1.13)%, P<0.01]. In the I-R group, the expression of mitochondrial ALDH2 was significantly reduced and that of Bcl-2 and Bax increased. Compared with the I-R group, the expression of mitochondrial ALDH2 was not significantly decreased in the D/IR group (P=0.11), while the expression of Bcl-2 and Bax was significantly increased. Compared with the I-R group, the D/I-R group had significantly increased Bcl-2/Bax ratio (0.44 vs 0.22, P=0.001).

Conclusion

Diltiazem reduces MIRI possibly by up-regulating the expression of mitochondrial Bcl-2 and down-regulating the expression of Bax. However, the therapeutic effect of diltiazem is not related with the gene and protein expression of mitochondrial ALDH2.

表1 目的基因引物序列
表2 心律失常评分方案
表3 各组大鼠再灌注左心室发展压比较(mmHg,±s
表4 各组大鼠再灌注30 min以及45 min左心室内压上升/下降最大速率(mmHg,±s
表5 各组大鼠再灌注室性心律失常发生率、时程及再灌注心律失常评分比较
图1 各组大鼠心脏染色切片
图2 各组大鼠心肌线粒体ALDH2、Bcl-2以及Bax蛋白表达
1
Chen YR, Nie SD, Shan W, et al. Decrease in endogenous CGRP release in nitroglycerin tolerance: role of ALDH-2 [J]. Eur J Pharmacol, 2007, 571(1): 44-50.
2
Krzesinski JM, Saint-Remy A. Essential hypertension, a complex trait [J]. Rev Med Liege, 2012, 67(5/6): 279-285.
3
Curtis MJ, Walker MJ. Quantification of arrhythmias using scoringsystems: an examination of seven scores in an in vivo model ofregional myocardial ischaemia [J]. Cardiovasc Res, 1988, 22(9): 656-665.
4
Fukuda S, Nakamura Y, Egi K, et al. Comparison of direct effects of clinically available vasodilators; nitroglycerin, nifedipine, cilnidipine and diltiazem, on human skeletonized internal mammary harvested with ultrasonic scalpel [J]. Heart Vessels, 2016, 31(10): 1681-1684.
5
张文彤, 闫洁. SPSS统计分析基础教程 [M]. 北京: 高等教育出版社, 2004: 289-292.
6
DiFabio J, Ji Y, Vasiliou V, et al. Role of Mitochondrial Aldehyde Dehydrogenase in Nitrate Tolerance [J]. Mol Pharmacol, 2003, 64(5): 1109-1116.
7
Frömmel J, Končitíková R, Kopečný D, et al. Oxidation of imidazole- and pyrazole-derived aldehydes by plant aldehyde dehydrogenases from the family 2 and 10 [J]. Chem Biol Interact, 2019, 304: 194-201.
8
Matsumoto A, Ito S, Wakamatsu K, et al. Ethanol induces skin hyperpigmentation in mice with aldehyde dehydrogenase 2 deficiency [J]. Chem Biol Interact, 2019, 302: 61-66.
9
张云盛, 张文娟. 线粒体乙醛脱氢酶2在心血管疾病中的研究进展 [J/CD]. 中华临床医师杂志(电子版), 2016, 10(20): 3112-3116.
10
张文丽, 张丹, 祝振忠. 乙醛脱氢酶2基因多态性与心血管疾病 [J]. 中国临床医生杂志, 2018, 46(4): 393-396.
11
Chen CH, Budas GR, Churchill EN, et al. Activation of aldehyde dehydrogenase-2 reduces ischemic damage to the heart [J]. Science, 2008, 321(5895): 1493-1495.
[1] 王泽华, 郭子瑊, 陈帅, 狄靖凯, 闫泽辉, 冯腾达, 毛兴佳, 向川. 线粒体质量控制在骨关节炎中的研究进展[J/OL]. 中华关节外科杂志(电子版), 2024, 18(02): 215-224.
[2] 周月惠, 江梦钰, 薛宇轩, 卫杨文祥, 凡一诺, 万子艺, 刘予豪, 陈镇秋, 周驰. 线粒体动力学相关蛋白影响破骨细胞分化机制探讨[J/OL]. 中华关节外科杂志(电子版), 2024, 18(01): 60-68.
[3] 江雅婷, 刘林峰, 沈辰曦, 陈奔, 刘婷, 龚裕强. 组织相关巨噬素3 保护肺血管内皮糖萼治疗急性呼吸窘迫综合征的机制研究[J/OL]. 中华危重症医学杂志(电子版), 2024, 17(05): 353-362.
[4] 钟雅雯, 王煜, 王海臻, 黄莉萍. 肌苷通过抑制线粒体通透性转换孔开放缓解缺氧/复氧诱导的人绒毛膜滋养层细胞凋亡[J/OL]. 中华妇幼临床医学杂志(电子版), 2024, 20(05): 525-533.
[5] 吴卫照, 肖贞, 袁转苹, 吴丹, 李源斌. MTO1基因变异致联合氧化磷酸化缺陷症10型患儿的临床和遗传学分析[J/OL]. 中华妇幼临床医学杂志(电子版), 2023, 19(06): 719-727.
[6] 张永博, 张亮, 陈浏阳, 戴睿, 孙华, 杨盛, 孟博, 彭晴. 线粒体与椎间盘退变[J/OL]. 中华损伤与修复杂志(电子版), 2023, 18(03): 265-269.
[7] 吴沛玲, 娄月妍, 张洪艳, 陈东方, 刘雪青, 赵丽芳, 薛姗, 蒋捍东. 线粒体相关基因在特发性肺纤维化中的分析[J/OL]. 中华肺部疾病杂志(电子版), 2024, 17(02): 178-184.
[8] 黄程鑫, 陈莉, 刘伊楚, 王水良, 赖晓凤. OPA1 在乳腺癌组织的表达特征及在ER阳性乳腺癌细胞中的生物学功能研究[J/OL]. 中华细胞与干细胞杂志(电子版), 2024, 14(05): 275-284.
[9] 张晟豪, 周杰, 姚鹏飞, 李长栋, 屈晓东, 南亚强, 曹丽. 雷公藤红素在创伤性脑损伤后继发性损伤中的作用及机制研究[J/OL]. 中华神经创伤外科电子杂志, 2024, 10(03): 132-140.
[10] 苗楠, 宗子钰. 脑出血后继发性脑损伤与线粒体相关机制的研究进展[J/OL]. 中华神经创伤外科电子杂志, 2024, 10(02): 107-111.
[11] 于伟伟, 张国高, 吴军, 胡俊, 黄一宁, 徐晶. 线粒体相关内质网膜相关线粒体功能障碍在阿尔茨海默病中的研究进展[J/OL]. 中华临床医师杂志(电子版), 2024, 18(02): 223-230.
[12] 闵志群, 苏杭. 广州地区人群ALDH2多态性分布特征与心脑血管疾病的相关性研究[J/OL]. 中华临床实验室管理电子杂志, 2024, 12(02): 97-102.
[13] 于乾雪, 廖学梅, 孙龙龙, 范梦莹, 蒋明超, 孟慧, 李瑞基. 线粒体功能障碍与卵巢早衰的研究进展[J/OL]. 中华诊断学电子杂志, 2023, 11(04): 283-288.
[14] 罗婷, 邱令智, 易东, 鄢华. 线粒体功能障碍与心血管疾病、缺血性脑卒中及慢性肾脏病关系的研究进展[J/OL]. 中华脑血管病杂志(电子版), 2024, 18(01): 60-63.
[15] 邱甜, 杨苗娟, 胡波, 郭毅, 何奕涛. 亚低温治疗脑梗死机制的研究进展[J/OL]. 中华脑血管病杂志(电子版), 2023, 17(05): 518-521.
阅读次数
全文


摘要