摘要
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 observe the protective effects of lycopene on myocardial cells through the establishment of the model of oxidatie stress damage induced by H_20_2 in myoeardium cells. Method： Neonatal rat eardiomyocytes were randomly divided into five groups： control, model （H_20_2） , H_20_2 ± lycopene （2.5 μmol · L-l） , H_20_2 ± lycopene （5 μmol·L-1） and H_20_2 ± lycopene （10 p, mol·L-1）. H_20_2 was used to induce oxidative stress injury in primary cultured cardiomyocytes of neonatal rats for 24 hours after incubated for 4 hours with lyeopene. Cell viability was detected by MTT assay. The content of lactate dehydrogenase （LDH） , creatine kinase （CK） in culture medium and malondialdehyde （MDA）, and the activities of superoxide dismutase （SOD） , catalase （CAT） in cell were detected after treated by lycopene with colorimetric technique at 24 hours. Observed the changes of cell nucleus shape and detected Caspase-3 expression of apoptosis proteins. Result： Compared with the normal group, model group could decrease viability of cardiomyocytes, the contents of LDH, CK and MDA in culture medium increased, and activities of SOD and CAT decreased, P 〈 0.01, and the characteristics of apoptotic cells presented, and Caspase-3 expression increased, P 〈 0.01. Compared with the model group, H_20_2 ± lyeopene （10 μmol·L-1） group could increase viability of cardiomyocytes （86.36±2.54）, （54. 15 ±10.97）%, P〈0.01, and the contents of LDH （319.53 ±23.48）, （495.27 ±31.57） U·L-1, P〈 0.01, CK （279.29±32.91）, （397.01±29.36） U·L-1, P〈0.01, andMDA （14.27±2.43）, （20.11± 2.09） μmol·g-1 P 〈0.01 in culture medium were decreased and activities of SOD （47.94 ± 2.64） （37.19±5.78） U·mg-1, P〈0.01, and CAT （34.95±4.67） , （16.35±4.84） U·mg-1, P〈0.01, were increased and improved nuclear morphology, and reduced Caspase-3 expression （ 1.40 ±0.20）, （ 1.94 ±0. 22） , P 〈 0.05. Conclusion： Lycopene exerted protective effects on oxidative stress injury induced by H_20_2 in cardiomyocytes of neonatal rats.
出处
《中国实验方剂学杂志》
CAS
北大核心
2014年第12期160-164,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81102288)
四川省教育厅重点项目(2013SZA0429)
