论文部分内容阅读
目的:通过建立H2O2诱导心肌细胞氧化应激损伤模型,观察番茄红素(lycopene)对心肌细胞的保护作用。方法:将原代培养的乳鼠心肌细胞随机分为正常组、模型组(H2O2)、番茄红素低剂量组(2.5μmol·L-1)、番茄红素中剂量组(5μmol·L-1)、番茄红素高剂量组(10μmol·L-1)。番茄红素与心肌细胞孵育4 h后,再加入H2O2,24 h后MTT法测定细胞存活率,比色法测定细胞培养上清液中乳酸脱氢酶(LDH)、肌酸激酶(CK)的含量和细胞超氧化物歧化酶(SOD)、过氧化氢酶(CAT)及丙二醛(MDA)含量,荧光电子显微镜下观察细胞核的形态变化,采用Western blot方法检测细胞凋亡相关蛋白半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)的变化。结果:与正常组比较,模型组心肌细胞存活率降低,心肌细胞LDH和CK释放量增加,SOD,CAT活性降低,MDA含量增加(P<0.01),呈现出凋亡细胞的特征,Caspase-3的表达增高(P<0.01)。番茄红素高剂量组与模型组比较,心肌细胞的存活率增加(86.36±2.54)%,(54.15±10.97)%,P<0.01,H2O2所致乳鼠心肌细胞LDH释放量降低(319.53±23.48),(495.27±31.57)U·L-1,P<0.01,CK释放量降低(279.29±32.91),(397.01±29.36)U·L-1,P<0.01,SOD活性增高(47.94±2.64),(37.19±5.78)U·mg-1,P<0.01,CAT活性增高(34.95±4.67),(16.35±4.84)U·mg-1,P<0.01,MDA含量降低(14.27±2.43),(20.11±2.09)μmol·g-1,P<0.01,细胞核形态改善,Caspase-3的表达降低(1.40±0.20),(1.94±0.22),P<0.05。结论:番茄红素对H2O2所致乳鼠心肌细胞氧化应激损伤有明显保护作用。
OBJECTIVE: To establish a model of H2O2-induced oxidative stress injury in cardiomyocytes and observe the protective effects of lycopene on cardiomyocytes. Methods: Primary cultured neonatal rat cardiomyocytes were randomly divided into normal group, model group (H2O2), low-dose group of lycopene (2.5μmol·L-1), middle dose of lycopene group (5μmol·L-1 ), High-dose lycopene group (10μmol·L-1). Lycopene was incubated with cardiomyocytes for 4 h, then H2O2 was added for 24 h, cell viability was measured by MTT assay, and lactate dehydrogenase (LDH), creatine kinase (CK) (SOD), catalase (CAT) and malondialdehyde (MDA) contents were observed. Morphological changes of the nucleus were observed under a fluorescence electron microscope. Western blot was used to detect the expression of apoptosis-related protein cysteine Changes of Caspase-3. Results: Compared with the normal group, the survival rate of myocardial cells in model group decreased, the release of LDH and CK, the activity of SOD and CAT, the content of MDA increased (P <0.01), the expression of Caspase-3 (P <0.01). Lycopene high dose group compared with the model group, the survival rate of myocardial cells increased (86.36 ± 2.54)%, (54.15 ± 10.97)%, P <0.01, H2O2-induced LDH release in neonatal rat cardiomyocytes decreased (319.53 ± 23.48 (495.27 ± 31.57) U · L-1, P <0.01, the release of CK was decreased by 279.29 ± 32.91 and 397.01 ± 29.36 U · L-1, P <0.01, and the activity of SOD was increased by 47.94 ± 2.64, (P <0.01), the activity of CAT increased (34.95 ± 4.67), (16.35 ± 4.84) U · mg-1, P <0.01, and the MDA content decreased (14.27 ± 2.43, 20.11 ± 2.09) μmol · g-1, P <0.01. The nuclear morphology was improved and the expression of Caspase-3 was decreased (1.40 ± 0.20), (1.94 ± 0.22), P <0.05. Conclusion: Lycopene has obvious protective effect on oxidative stress injury of neonatal rat cardiomyocytes induced by H2O2.