摘要
目的探讨阿魏酸预处理对晚期糖基化终末产物(AGEs)诱导H9c2心肌细胞氧化应激损伤的保护作用。方法大鼠H9c2心肌细胞分为对照组、模型组和实验组。实验组予以20. 0μmol·L^-1阿魏酸20μL预处理,模型组予以等量0. 9%NaCl预处理2 h,然后均加入100 mg·L^-1AGEs共培养24 h;对照组细胞则以等量0. 9%NaCl处理。给药24 h后,用活细胞计数(CCK-8)法检测H9c2心肌细胞存活率,以2,7-二氯荧光素二乙酸盐(DCF-DA)荧光染色检测H9c2心肌细胞内活性氧(ROS)水平,用试剂盒检测超氧化物歧化酶(SOD)、丙二醛(MDA)水平;给药48 h后,用蛋白免疫印迹法检测H9c2心肌细胞转录因子NF-E2相关因子2(Nrf-2)、血红素氧合酶-1(HO-1)蛋白表达情况。结果对照组、模型组及实验组H9c2心肌细胞存活率分别为(95. 84±1. 63)%,(72. 57±4. 87)%,(82. 14±4. 43)%,细胞内ROS相对荧光强度值分别为3. 14±0. 85,26. 12±3. 64,13. 01±2. 55;心肌细胞中MDA分别为(33. 26±5. 12),(68. 15±4. 29),(42. 15±3. 69)μmol·mg^-1Pro;SOD分别为(46. 59±6. 05),(32. 05±6. 99),(42. 15±5. 02) U·mg^-1Pro;Nrf-2蛋白相对表达量分别为0. 89±0. 15,0. 22±0. 06,0. 63±0. 08;HO-1蛋白相对表达量分别为2. 63±0. 52,0. 86±0. 18,1. 89±0. 33。模型组分别与对照组和实验组比较,差异均有统计学意义(均P <0. 05).结论阿魏酸预处理通过调控Nrf-2、HO-1蛋白表达量,抑制AGEs诱导H9c2心肌细胞氧化应激反应,减轻心肌损伤。
Objective To investigate the protective effect of ferulic acid( FA) pretreatment on oxidative stress injury induced by advanced glycation end products( AGEs) in H9c2 cardiomyocytes. Methods Rat H9c2 cardiomyocytes were assigned to control group,model group and test group. The test group was given pretreatment with 20. 0 μmol·L^-1 FA 20 μL,and the model group was given pretreatment with equal amount of 0. 9% NaCl for 2 h,and then treated with 100 mg·L^-1 of AGEs for 24 h,while the control group was pretreated with equal amount of 0. 9% NaCl. After 24 h of administration,viability of H9c2 myocardial cells was measured by cell count kit( CCK)-8 assay,reactive oxygen species( ROS) was detected by 2,7-dichlorofluorescein diacetate( DCF-DA) fluorescence staining,malondialdehyde( MDA),superoxide dismutase( SOD) in H9c2 myocardial cells were measured by assay kits.At 48 h after treatment,NF-E2-related factor 2( Nrf-2) and hemeoxygenase-1( HO-1) protein expression in H9c2 myocardial cells were detected by Western blotting. Results Of the control group,model group and test group,the survival rates of H9c2 myocardial cells were( 95. 84 ± 1. 63)%,( 72. 57 ± 4. 87)%,( 82. 14 ± 4. 43)%;the relative fluorescence intensities of ROS in the cells were 3. 14 ± 0. 85,26. 12 ± 3. 64,13. 01 ± 2. 55;the levels of MDA were( 33. 26 ± 5. 12),( 68. 15 ± 4. 29),( 42. 15 ± 3. 69)μmol·mg^-1 Pro;the levels of SOD were( 46. 59 ± 6. 05),( 32. 05 ± 6. 99),( 42. 15 ± 5. 02) U·mg^-1 Pro;the relative expression of Nrf-2 protein was 0. 89 ± 0. 15,0. 22 ± 0. 06,0. 63 ± 0. 08,respectively;the relative expression of HO-1 protein was 2. 63 ± 0. 52,0. 86 ± 0. 18,1. 89 ± 0. 33. There were statistically significances between the model group and the control/test group( all P < 0. 05).Conclusion FA pretreatment can inhibit AGEs-induced oxidative stress in H9c2 cardiomyocytes and alleviate myocardial injury by regulating the expression of Nrf-2 and HO-1 protein.
作者
李玲
谢峥
万有才
LI Ling;XIE Zheng;WAN You-cai(Department of Pharmaceutical,Second People’s Hospital of Jingmen City, Jingmen 448000, Hubei Province, China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2019年第14期1446-1448,共3页
The Chinese Journal of Clinical Pharmacology
关键词
心肌损伤
阿魏酸
晚期糖基化终末产物
氧化应激
myocardial injury
ferulic acid
advanced glycation end products
oxidative stress
