摘要
目的:研究神经调节蛋白1(NRG-1)对高糖损伤的心肌细胞的保护作用及其机制。方法:培养大鼠胚胎期心脏组织H9c2心肌细胞,进行分组处理。分别为对照组(不加任何诱导药物处理24 h)、高糖作用组(HG组,用含终浓度33 mmol/L的葡萄糖培养基培养心肌细胞处理24 h)、不同浓度NRG-1作用组(分别在含有33 mmol/L高浓度葡萄糖的培养基中同时加入10、50、250 nmol/L的NRG-1培养心肌细胞处理24 h,设定为N1、N2、N3组)。细胞计数(CCK-8)方法检测各组心肌细胞存活率;流式细胞术检测各组心肌细胞胞内活性氧(ROS)水平和细胞凋亡情况;同时测定各组心肌细胞中肌酸激酶(CK)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)活力和丙二醛(MDA)含量;蛋白免疫印迹(Western-blot)方法检测各组心肌细胞中NRG-1的受体,即表皮生长因子受体-2(Erb B2)和表皮生长因子受体-4(Erb B4)的表达;原位缺口末端标记法(Tunel)染色检测2型糖尿病心肌病模型大鼠经NRG-1处理后的心肌细胞凋亡。结果:N1组至N3组心肌细胞的存活率由(63.33±3.56)%上升至(85.88±4.55)%;心肌细胞中ROS含量由(33.57±4.23)%下降至(15.88±4.55)%;细胞凋亡率由(36.44±4.86)%下降至(14.77±4.21)%;心肌细胞中Erb B2和Erb B4受体表达随之分别由(0.26±0.04)、(0.39±0.03)上升至(0.84±0.03)、(0.72±0.04),与HG组比差异均有统计学意义(P<0.05);CK、LDH酶活力和MDA含量逐渐下降而SOD活性逐渐回升,与HG组比差异均有统计学意义(P<0.05)。NRG-1作用的2型糖尿病心肌病模型大鼠心肌组织细胞凋亡也显著下降。结论:NRG-1能够提升高糖作用下心肌细胞存活率,减轻高糖对心肌细胞氧化应激损伤和细胞凋亡,可能通过与心肌细胞中Erb B2/Erb B4受体结合发挥作用。
Objective： To study the protective roll of neuregulin-1 （NRG-1） on high glucose caused myocardial cell injury in rat’s embryo H9c2 myocardial cells with its mechanism. Methods： Cultured rat’s embryo H9c2 myocardial cells were divided into 5 groups：①Control group,②High glucose （HG） group, containing glucose 33 mmol/L,③HG＋NRG-1 10 nmol/L （N1） group,④HG＋NRG-1 50 nmol/L （N2） group and⑤HG＋NRG-1 250 nmol/L （N3） group. All cells were treated for 24 hours. Myocardial cell survival rate was measured by CCK-8 method, intracellular reactive oxygen species （ROS） level and the apoptosis rate were detected by lfow cytometry, enzymes activities of CK, LDH, SOD and MDA content were examined, proteins expressions of NRG-1 receptor as ErbB2 and ErbB4 were assessed by Western blot analysis. NRG-1 treated myocardial cell apoptosis in type II diabetic cardiomyopathy rats was observed by Tunel staining. Results： Compared with HG group, from N1 group to N3 group, myocardial cell survival rates were increased from （63.33±3.56） %to （85.88±4.55） %, ROS levels decrease form （33.75±4.23） % to （15.88±4.55） %, apoptosis rates reduced from （36.44±4.86） % to （14.77±4.21） %, receptor expressions of ErbB2 was elevated from （0.26±0.04） to （0.84±0.03） and ErbB4 was elevated from （0.39±0.03） to （0.72±0.04）, allP〈0.05; enzymes activities of CK, LDH and MDA content were gradually decreased and SOD activity was gradually increased, allP〈0.05. NRG-1 treated myocardial cell apoptosis in type II diabetic cardiomyopathy rats was also obviously reduced. Conclusion： NRG-1 could increase the survival rate and reduce the oxidative stress injury and apoptosis of cultured rat’s embryo H9c2 myocardial cells in HG condition which might be related to NRG-1 binding to ErbB2/ErbB4 molecules in the cells.
作者
罗明雄
陈小丹
魏玲
LUO Ming-xiong CHEN Xiao-dan WEI Ling(Department of Cardiology, Suzhou Wuzhong People＇s Hospital, Suzhou （215128）, Jiangsu, China)
出处
《中国循环杂志》
CSCD
北大核心
2016年第9期902-907,共6页
Chinese Circulation Journal
