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103μg/L组、104μg/L组和105μg/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 myo- cardial 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 p,g/L, 104 p,g/L and 105 p^g/L CsA ）. Twenty-four h after the treatment, cellular morphological changes were observed by inverted phase contrast microscope and electron microscope, respectively. MTF 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. Vaeuolizations were observed in the endochylema of myocardial cells in CsA-treated groups. Uhrastructure of eardiomyocytes 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 mecha- nism of which may be associated with the damage of lipid peroxidation.
出处
《心脏杂志》
CAS
2010年第5期684-687,共4页
Chinese Heart Journal
基金
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)
关键词
环孢霉素A
心肌细胞
心肌损伤
乳鼠
cyctosporine A
myocardial cells
myocardial cell injury
neonatal rat
