«上一篇/Previous Article|本期目录/Table of Contents|下一篇/Next Article»

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:

2016年第5期

页码:

512-517

栏目:

基础研究

出版日期:

2016-04-25

文章信息/Info

Title:

Protective effect of quercetin on secondary cardiomyocyte injury induced by mechanical trauma

作者:

杨学颖¹; 2; 王卓润¹; 李秀杰¹; 景泽昊¹; 马 硕¹; 于德钦¹; 李树壮¹

（1.大连医科大学生理学教研室，辽宁 大连 116044；
2.解放军沈阳军区大连疗养院桃园疗区，辽宁 大连 116013）

Author(s):

YANG Xue-ying¹; 2;  WANG Zhuo-run¹;  LI Xiu-jie¹;  JING Ze-hao¹;  MA Shuo¹;  YU De qin¹;  LI Shu-zhuang¹

(1.Department of Physiology, Dalian Medical University, Dalian 116044, Liaoning, China；
2.Taoyuan Sanatorium, Shenyang Military Area Command, Dalian 116013, Liaoning, China)

关键词:

槲皮素; 心脏保护; 氧化应激

Keywords:

quercetin;  heart injuries;  oxidative stress

分类号:

R284

DOI:

-

文献标识码:

A

摘要:

目的 观察槲皮素对机械性创伤（MT）造成大鼠继发性心肌细胞及心脏功能损伤的保护效果，同时探究其作用机制。 方法 实验分为在体实验和离体实验。将120只成年雄性SD大鼠按随机数字表法分为4组，分别为正常组、创伤组、创伤+槲皮素和创伤+溶剂组。先后应用小动物定量创伤仪制备大鼠MT模型；经右颈总动脉向左心室内插管，记录左心室舒缩压力变化，检测大鼠心功能，初步研究槲皮素对心脏的保护作用；通过MTT比色法检测槲皮素对创伤诱导的H9c2细胞存活率的影响，确定槲皮素对心脏的保护效果；在槲皮素的保护机制研究中，TUNEL染色后计算心肌细胞凋亡指数，以评估心肌细胞的凋亡状况；通过酶标仪检测H9c2细胞在加入槲皮素前后产生活性氧(ROS)自由基的量的变化；激光共聚焦显微镜下观察测定经Fluo-4AM标记的心肌细胞内Ca2+的浓度变化，从而初步探讨槲皮素发挥保护作用的机制。结果 ①心功能指标LVDP、+dP/dtmax 、-dP/dtmax显示，创伤组、创伤+溶剂组与正常组比较均明显下降，而创伤+槲皮素组较创伤组升高（P<0.01）。②MTT检测发现，在一定的浓度范围内，槲皮素对H9c2细胞没有细胞毒性作用，并且可明显阻碍创伤血清对H9c2细胞的损伤。③MT后通过对正常组、创伤组、创伤+槲皮素组和创伤+溶剂组心肌细胞凋亡指数的检测，发现创伤+槲皮素组与创伤组比较凋亡指数明显降低(P<0.01)。④通过对H9c2细胞内ROS的检测，发现槲皮素能够减低MT后细胞内ROS的产生。⑤通过对心肌细胞内钙浓度变化的检测，发现MT后Ca2+浓度上升，给予一定浓度槲皮素后Ca2+浓度下降。结论 在一定的浓度范围内，槲皮素能够降低MT后产生的ROS自由基，抑制Ca2+内流，继而降低心肌细胞凋亡,改善MT后心脏功能，发挥心脏保护作用。

Abstract:

AIM To observe the effect of quercetin on secondary cardiomyocyte apoptosis and cardiac dysfunction induced by mechanical trauma in rats and discuss its mechanism. METHODSThe experiment was conducted in vivo and in vitro. First, 120 adult male Sprague Dawley rats were randomly divided into four groups: sham group, trauma group, trauma+qucercetin group and trauma+vehicle group for experiments in vivo. We used Noble-Collip drum to prepare the mechanical trauma model of rats. We then recorded the changes of the left ventricular diastolic pressure with the intubation from the right common carotid artery to the left ventricle to identify the protective effect of quercetin on rat cardiac function. MTT assay was used to evaluate the effects of quercetin on the survival rate of H9c2 cells induced by trauma plasma. Using TUNEL staining, we assessed the apoptosis degree of cardiomyocytes. Reactive oxygen species (ROS) produced by H9c2 cells before and after the administration of quercetin was detected by microplate reader to substantiate the protective effects of quercetin. We used laser scanning confocal microscope to observe the cardiomyocytes marked by Fluo-4AM to determine the changes of intracellular Ca2+. RESULTSLVDP, +dP/dtmax and -dP/dtmax in trauma group and trauma+vehicle group significantly decreased, and those in trauma+quercetin group increased compared with those in sham group (P<0.01). Under tested concentrations, quercetin had no toxicity to H9c2 cells and could obviously inhibit the damage of trauma plasma. The apoptosis index of cardiomyocytes in the trauma+quercetin group was lower than that in the trauma group (P<0.01). By detecting the oxide in H9c2 cells, we observed that quercetin reduced MT-induced overproduction of ROS. The concentration of intracellular calcium increased when cultured with trauma plasma and after given a certain concentration of quercetin, the concentration of intracellular calcium decreased. CONCLUSIONUsed at an appropriate concentration, quercetin can reduce the overproduction of ROS induced by MT and inhibit MT-initiated Ca2+ influx. Quercetin can also inhibit myocardial apoptosis, improve heart function after MT and exert some protective effect on the heart.

参考文献/References

[1]Ismailov RM,Ness RB,Weiss HB et al.Trauma associated with acute myocardial infarction in a multi-state hospitalized population[J].Int J Cardiol,2005,105(2):141-146.
[2]Yan Z,Liang F,Guo L,et al.Myeloperoxidase increased cardiomyocyte protein nitration in mice subjected to nonlethal mechanical trauma[J].Biochem Biophys Res Commun,2010,393(3):531-535.
[3]Cash TP,Pan Y,Simon MC.Reactive oxygen species and cellular oxygen sensing[J].Free Radic Biol Med,2007,439(9):1219-1225.
[4]Buonocore G,Perrone S,Tataeanno ML.Oxygen toxicity:chemistry and biology of reactive oxygen species[J].Semin Fetal Neonatal Med,2010,15(4):189-190.
[5]Circu ML,Aw TY.Reactive oxygen species,cellular redox systems,and apoptosis[J].Free Radic Biol Med,2010,48(6):749-762.
[6]Murakami A,Ashida H,Terao J.Multitargeted cancer prevention by quercetin[J]. Cancer lett,2008,269(2):315-325.
[7]Shutenko Z,Heney Y,Pinard E,et al.Influence of the antioxidant quercetin in vivo on the level of nitric oxide detemined by electro paramagnetic resonance in rat brain during global ischemia and reperfusion[J].Biochem Pharmacol,1999,57:199.
[8]Davis JM,Murphy EA,Carmichael MD.Effects of the dietary flavonoid quercetin upon performance and health[J].Curr Sports,Med Rep,2009,8(4):206-213.
[9] 刘 宣,吕广艳,吕国枫,等.自制小动物定量创伤仪对大鼠心肌组织损伤的观察[J].心脏杂志,2012,24(5):583-587.
[10]郭 丽,王晓樑,赵焕新,等.机械性创伤致心肌细胞凋亡发生的时间规律[J].中国应用生理学杂志,2009,25(3):352-354.
[11]石松菁.多器官功能衰竭救治进展[J].中华急诊医学杂志,2014,23(8): 844-846.
[12]Oghlakian G,Maldjian P,Kaluski E, et al.Acute myocardial infarction due to left anterior descending coronary artery dissection after blunt chest trauma[J].Emerg Radiol,2010,17(2):149-151.
[13]曹成章,燕 子,温永金,等.机械创伤后肾动脉内皮依赖性舒张功能改变与迟发性心功能不全的关系[J].中华创伤杂志,2013,29(4):348-352.
[14]Tao L,Liu HR,Gao F,et al.Mechanical traumatic injury without circulatory shock causes cardiomyocyte apoptosis:role of reactive nitrogen and reactive oxygen species[J].Am J Physiol Heart Circ Physiol,2005,288(6):H2811-H2818.
[15]Li S,Jiao X,Tao L,et al.Tumor necrosis factor-alpha in mechanic trauma plasma mediates cardiomyocyte apoptosis[J].Am J Physiol Heart Circ Physiol,2007,293(3):H1847-H1852.
[16]Zhang L,Xiao Y,He J.Cocaine and apoptosis in myocardial cells[J].Anat Rec,1999,257(6):208-216.

备注/Memo

备注/Memo:

收稿日期：2015-09-14.
基金项目：国家自然科学基金项目资助(30971065)；大连市科技计划项目资助(2012E12SF074)；辽宁省教育厅基金项目资助(2009A194)
通讯作者：李树壮，教授，主要从事循环系统的损伤与保护研究 Email:shuzhuangli@126.com
作者简介：杨学颖，副主任护师 Email:yangxueying6@sina.com 共同第一作者：王卓润，本科生 Email:1241004187@qq.com

常用功能

更新日期/Last Update: 2016-04-13