叔丁基对苯二酚拮抗心肌细胞氧化应激损伤机制研究  

Effects of tert-butylhydroquinone(t BHQ) against oxidative stress injury of H9c2 myocardial cells

作　　者：刘辉[1] 刘红阳[2] 姜一农[3] 常栋[3] 李楠[1] 张彧[1] 

机构地区：[1]大连医科大学附属第一医院急诊科,大连116011 [2]大连医科大学附属第一医院心脏重症监护科,大连116011 [3]大连医科大学附属第一医院心内科,大连116011

出　　处：《中国实用内科杂志》2015年第8期704-707,共4页Chinese Journal of Practical Internal Medicine

基　　金：国家自然科学基金(81200136)

摘　　要：目的探讨叔丁基对苯二酚(t BHQ)对化学性低氧模型剂氯化钴(Co Cl2)诱发H9c2心肌细胞氧化应激损伤的影响及其作用机制。方法将细胞分成4组:正常对照组、Co Cl2组(500μmol/L Co Cl2)、DMSO(二甲基亚砜)+Co Cl2(0.005%DMSO+500μmol/L Co Cl2)组、t BHQ+Co Cl2(50μmol/L t BHQ+500μmol/L Co Cl2)组;MTT法检测细胞生长活性;DCFH-DA探针测定细胞内ROS含量;Western Blot检测细胞内cleaved-caspase-3和Nrf2及其调控的II相解毒酶NAD(P)H:醌氧化还原酶1(NQO1)和抗氧化酶γ-谷氨酰半胱氨酸连接酶(GCLC)蛋白的表达;应用Real-time PCR检测NQO1和GCLC m RNA表达。结果 Co Cl2组与正常对照组相比,细胞活性明显降低,细胞内的cleaved-caspase-3蛋白表达显著升高,ROS含量增高;DMSO+Co Cl2组与Co Cl2组无差异;t BHQ预处理能够显著降低Co Cl2对H9c2心肌细胞氧化应激损伤。与Co Cl2组相比,t BHQ预处理能够显著增加细胞内的Nrf2蛋白水平,且细胞内NQO1和GCLC的m RNA及蛋白表达均明显高于Co Cl2组。结论 t BHQ能够诱导H9c2心肌细胞内Nrf2的蛋白表达,进而上调其下游II相解毒酶NQO1和抗氧化酶GCLC的转录活性及蛋白表达,拮抗Co Cl2诱发的化学性缺氧对H9c2心肌细胞的氧化应激损伤。Objective To invegtigate the effect of tert-butylhydroquinone （tBHQ） on the oxidative stress damages induced by chemical hypoxia-mimetic agent （cobalt chloride, COC12） in H9c2 myocardial ceils as well as its mechanisms. Methods The H9c2 cells were divided into 4 groups： normal control group, CoC12 group （induced by 500μmol/L COC12）, DMSO＋CoC12 group （induced by 0.005%DMSO ＋ 500 μmol/L COC12）, tBHQ＋CoC12 group （induced by 50 mol/L tBHQ.＋500 mol/L COC12）. The cell growth viability was detected by MTT assay, and the content of ROS in cells was measured by the DCFH-DA probe. The protein expression of cleaved- caspase-3, Nrf2 and its regulation of phase Ⅱdetoxifying enzyme NAD （P） H： quinone oxidoreductase 1 （NQO1） and antioxidant enzymes y-glutamate cysteine ligase catalytic subunit （GCLC） were determined with Western Blot. The mRNA expression ofNQO1 and GCLC were measured by real-time polymerase chain reaction （PCR）. Results In comparison with those in the normal control group, the cell viability decreased markedly, and the expression of cleaved-caspase-3 protein and ROS content increased significantly in the COC12 group. There were no significant differences between DMSO＋CoC12 group and COC12 group. Pretreatment with tBHQ could markedly inhibit CoC1z induced-oxidative stress damages in H9c2 cells. As compared with CoC1z group, tBHQ.pretreatment remarkably increased the level of Nrf2 protein, and the expression levels of NQO 1 and GCLC mRNA and protein were significantly elevated in the COC12 group. Conclusion TBHQmay protect H9c2 myocardial cells against CoC12-induced oxidative stress injuries by inducing the expression of Nrf2 protein expression, activating the transcription of the downstream.phase II detoxifying enzymes NQO 1 and antioxidant enzymes GCLC and up-regulating the protein expression of NQO 1 and GCLC in H9c2 cells.

关 键 词：叔丁基对苯二酚 氯化钴 NRF2 心肌细胞 氧化应激 

分 类 号：R544.1[医药卫生—心血管疾病]