铁离子螯合剂去铁敏在大鼠心肌细胞原代培养中对心肌细胞的保护作用  

Protective Effect of Deferoxamine on Primary Culture of Cardiomyocytes in Rats

作　　者：王耀晟[1] 邹循锋[2] 鲍晓明[1] 黄晓[1] 李萍[1] 程晓曙[1] 

机构地区：[1]南昌大学第二附属医院心血管内科,江西省南昌市330006 [2]杜伊斯堡-艾森大学艾森医学院生物化学研究所,艾森德国45147

出　　处：《中国循环杂志》2009年第5期387-390,共4页Chinese Circulation Journal

基　　金："十一五"国家科技支撑计划项目(2008BA168B00)

摘　　要：目的:在心肌细胞培养液中添加去铁敏,以对抗原代培养操作过程所产生的心肌细胞损伤,并探讨其保护作用。方法:在常规新生SD乳鼠心肌细胞原代培养液中按照不同浓度添加去铁敏。抗α-肌动蛋白免疫组化法鉴定心肌细胞。心肌细胞分为对照组(不添加去铁敏);5μmol/L去铁敏组;15μmol/L去铁敏组;25μmol/L去铁敏组。MTT法检测心肌细胞存活率、乳酸脱氢酶活性检测、TUNEL法检测心肌细胞凋亡率评价各组心肌细胞的损伤程度、生存率及凋亡情况。结果:①5μmol/L去铁敏组(69.6±5.4IU/L)、15μmol/L去铁敏组(35.5±3.3 IU/L)和25μmol/L去铁敏组(51.0±4.3 IU/L)的乳酸脱氢酶水平均比对照组(76.3±6.1 IU/L)低,差异均有统计学意义(P<0.05～0.01)。15μmol/L去铁敏组乳酸脱氢酶水平明显低于5μmol/L去铁敏组和25μmol/L去铁敏组,差异均有统计学意义(P<0.05～0.01)。②15μmol/L去铁敏组(91.28±2.57)%和25μmol/L去铁敏组(86.03±4.66)%心肌细胞生存率均比对照组(83.13±5.26)%高,差异均有统计学意义(P<0.05～0.01)。且15μmol/L去铁敏组心肌细胞生存率明显高于5μmol/L去铁敏组(84.19±3.36)%和25μmol/L去铁敏组,差异均有统计学意义(P<0.05～0.01)。③5μmol/L去铁敏组(7.11±0.93)、15μmol/L去铁敏组(3.10±1.26)和25μmol/L去铁敏组(5.03±0.82)心肌细胞凋亡指数均比对照组(8.06±1.51)低,差异均有统计学意义(P<0.05～0.01)。15μmol/L去铁敏组心肌细胞凋亡指数低于5μmol/L去铁敏组和25μmol/L去铁敏组,差异均有统计学意义(P<0.05～0.01)。结论:通过在心肌细胞培养液中加入适量去铁敏,能有效对抗原代培养操作过程带来的心肌细胞损伤,起到保护作用。Objective ： To investigate the effect of adding deferoxamine （DFO） in cardiomyocytes cultured medium for protecting cells against the injury from operative procedure of primary cuhured cardiomyocytes in rats. Methods： Primary cultured neonatal rats＇ cardiomyocytes were divided into 4 groups. Control group, no DFO was added; DFO 5 μmol/L group; DFO 15 μmol/L group and DFO 25 μmol/L group. DFO was added into the cultured medium in different doses respectively. Cardiomyocytes were identified by anti-α-actin immunohistochemical staining. The injury degree, survival rate and apoptosis rate were evaluated by methyl thiazolyl tetrazolium （MTT） assay,lactate dehydrogenase （LDH） activity detection and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） analysis respectively. Results： ①LDH leakage from cardiomyocytes in all DFO groups was lower than that in Control group （P 〈 0.01 ）. LDH leakage in DFO 15 μmol/L group was the lowest, 35.5 ± 3.3 IU/L （ 15 μmol/L） vs. 69. 6 ± 5.4 IU/L （ 5μmol/L） vs. 51.0 ± 4. 3 IU/L（25 μmol/L） ,P 〈 0. 01. ②Survival rates of cardiomyocytes in DFO 15 μmol/L and DFO 25 μmol/L groups were higher as compared with Control group （ P 〈 0. 05 ）. Survival rate in DFO 15 μmol/L group was the highest, 91.28 ± 2. 57% （15 μmol/L） vs. 84. 19 ± 3.36% （5μmol/L）vs. 86. 03 ± 4. 66% （25 μmol/L） ,P 〈 0. 01. ③Apoptosis rates of cardiomyocytes in all DFO groups were significantly reduced than that in Control group （ P 〈 0. 01 ）. Apoptosis rate in DFO 15 μmol/L group was the lowest,3.10 ± 1.26（ 15 μmol/L） vs. 7. 11 ±0. 93 （5 μmol/L） vs. 5.03 ±0. 82（25μmol/L） ,P 〈0. 01. Conclusion ： Appropriate dose of DFO in cultured medium could effectively protect the operative injury of cardiomyocytes in primary culture procedure.

关 键 词：去铁敏 心肌细胞 原代培养 凋亡 

分 类 号：R541[医药卫生—心血管疾病]