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|本期目录/Table of Contents|

环孢霉素A对乳鼠心肌细胞结构及功能的影响

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2010年第5期
页码:
684-687
栏目:
基础研究
出版日期:
2010-06-22

文章信息/Info

Title:
Effects of cyclosporine A on structure and function of cultured myocardial cells of neonatal rats
作者:
曹滢1; 胡晶2; 傅羽3; 于金凤4; 尹新华3
1.航空工业中心医院心内科，北京 100012；哈尔滨医科大学：2.附属第三医院心内科， 3.附属第一医院心内科；4.哈尔滨市胸科医院心内科,黑龙江 哈尔滨 150000
Author(s):
CAO Ying1;  HU Jing2;  FU Yu3;  YU Jin-feng4;  YIN Xin-hua3
1.Department of Cardiology, Center Hospital of Aviation Industry, Beijing 100012, China; 2.Department of Cardiology, Third Affiliated Hospital, 3.Department of Cardiology, First Affiliated Hospital, Harbin Medical University, Harbin 150000, Heilongjiang,
关键词:
环孢霉素A; 心肌细胞; 心肌损伤; 乳鼠
Keywords:
cyclosporine A;  myocardial cells;  myocardial cell injury;  neonatal rat
分类号:
R979.5
DOI:
-
文献标识码:
A
摘要:
目的: 观察环孢霉素A(cyclosporine A，CsA)对体外培养的乳鼠心肌细胞结构及功能的影响，探寻CsA诱导心肌细胞损伤的机制。方法: 采用酶消化法体外培养原代乳鼠心肌细胞，用100 ml/L DMEM培养基培养48 h后，随机分为对照组和CsA干预组。CsA干预组根据CsA的终浓度又分为103 μg/L组、104 μg/L组和105 μg/L组。分别于给CsA后24 h，在光镜及电镜下观察心肌细胞形态结构的变化。用MTT比色法检测细胞数量的变化，于给CsA后24 h及48 h，分别用比色法和硫代巴比妥酸（TBA）法检测细胞上清液中乳酸脱氢酶（LDH）及丙二醛（MDA）含量的变化。结果: 光镜下可见CsA干预组心肌细胞的数量减少，形态欠规则，胞浆中可见空泡，跳动频率减低，且随着用药浓度的增大而变化明显。电镜下可见CsA干预组心肌细胞线粒体肿胀、空泡样变，且随着浓度的增大而逐渐加重。MTT比色法检测显示，CsA干预组的细胞数量减少及活力降低（P<0.05）；而LDH及MDA的水平与对照组比明显升高（P<0.05，P<0.01），且二者随着CsA浓度的增大及时间的延长而差别显著（P<0.05）。结论: CsA对体外培养的心肌细胞有明显的损伤作用，且呈时间和剂量依赖性，可能与其致线粒体损伤及脂质过氧化有关。
Abstract:
AIM: To observe the effect of cyclosporine A (CsA) on the morphology and function of myocardial cells in vitro and to study the mechanism underlying CsA-induced injury in myocardial cells. METHODS: Myocardial cells of neonatal rats were cultured for 48 h with 100 ml/L DMEM and were randomly divided into four groups: normal control group and CsA treated groups (with 103 μg/L, 104 μg/L and 105 μg/L CsA). Twenty-four h after the treatment, cellular morphological changes were observed by inverted phase contrast microscope and electron microscope, respectively. MTT was adopted to measure the cell numbers. Lactate dehydrogenase (LDH) levels and malondialdehyde (MDA) levels in the supernatants of the cultured myocardial cells in each group were detected by colorimetric method and TBA 24 h and 48 h after treatment. RESULTS: The numbers and beats of myocardial cells in CsA-treated groups signigficantly decreased compared with those in control group. Vacuolizations were observed in the endochylema of myocardial cells in CsA-treated groups. Ultrastructure of cardiomyocytes was seriously damaged and mitochondrial swelling denaturation was found in CsA- treated groups. The cell numbers and activities in CsA-treated groups were all lower than those in control group (P<0.05, P<0.01). LDH and MDA levels in CsA-treated groups were higher than those in control group at 24 h and 48 h after treatment in a significant concentration- and time-dependent manner (P<0.05). CONCLUSION: CsA induces myocardial cell injury in a dose- and time-dependent manner, the mechanism of which may be associated with the damage of lipid peroxidation.

参考文献/References

［1］Rezzani R, Rodella L, Dessy C, et al. Changes in Hsp90 expression determinethe effects of cyclosporine a on the NO pathway in rat myocardium［J］. FEBS Lett, 2003, 552(2-3):125-129.

［2］常连庆，谢晓华，陈雯，等. 环孢霉素A对高血压大鼠心肌组织形态和结构的影响［J］ . 中国临床康复, 2005, 9(7):38-40.

［3］Stacchiotti A, Bonomini F, Lavazza A, et al. Adverse effects of cyclosporine A on HSP25, alpha B-crystallin and myofibrillar cytoskeleton in rat heart［J］. Toxicolog, 2009, 262(3):192-198.

［4］支继新，李学奇，尹新华，等. 环孢霉素A对大鼠心肌结构及心脏功能的影响［J］ . 中国地方病学杂志, 2006， 25(3):260-263.

［5］李扬，于金凤，张朝颖，等. 环孢霉素A诱导的心肌损伤与NO和NOS的关系［J］ . 哈尔滨医科大学学报, 2009, 43(3):241-254.

［6］Oka N, Wang L, Mi W, et al. Cyclosporine A prevents apoptosis-related mitochondrial dysfunction after neonatal cardioplegic arrest［J］. J Thorac Cardiovasc Surg, 2008, 135(3):123-130.

［7］Tharakan B, Holder-Haynes JG, Hunter FA, et al. Cyclosporine A prevents vascular hyperpermeability after hemorrhagic shock by inhibiting apoptotic signaling［J］. J Trauma, 2009, 66(4):1033-1039.

[8] Rezzani R, Rodella LF, Fraschini F, et al. Melatonin delivery in solid lipid nanoparticles: prevention of cyclosporine A induced cardiac damage［J］. J Pineal Res, 2009, 46(3):255-261.

备注/Memo

备注/Memo:
收稿日期：2009-09-07.基金项目：国家科学自然基金资助（30872387）；高等学校博士学科点专项科研基金资助（200802260006） 通讯作者：尹新华，主任医师，主要从事免疫抑制剂毒副作用及支架再狭窄研究Email:harbin0910@yahoo.com 作者简介：曹滢，住院医师，硕士生Email:caoying361@yahoo.com.cn
更新日期/Last Update: 2010-06-22