神经调节蛋白1对高糖损伤的心肌细胞的保护作用及其机制

发布时间：2018-04-24 04:32

本文选题：神经调节蛋白- + 肌细胞　；参考：《中国循环杂志》2016年09期

【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(P0.05);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(P0.05)。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。
[Abstract]:Aim: to study the protective effect and mechanism of neuroregulatory protein 1 (NRG-1) on hyperglycemic cardiomyocytes. Methods: H9c2 cardiomyocytes from rat embryonic heart were cultured and treated in groups. The rats were divided into two groups: control group (without any induced drugs for 24 h), high glucose group (HG group), and high glucose group (HG group). Cardiomyocytes were cultured on glucose medium containing final concentration of 33 mmol/L for 24 h. The cardiomyocytes were treated with different concentrations of NRG-1 for 24 h. The cultured cardiomyocytes were treated with NRG-1 containing 33 mmol/L high concentration glucose for 24 h. The survival rate of cardiac myocytes in each group was detected by CCK-8 method, the intracellular reactive oxygen species (Ros) level and apoptosis of cardiac myocytes in each group were detected by flow cytometry. At the same time, the activities of creatine kinase (CK), lactate dehydrogenase (LDH), superoxide dismutase (SOD) and malondialdehyde (MDA) in cardiac myocytes were measured, and the receptors of NRG-1 in cardiac myocytes were detected by Western blot. The expression of epidermal growth factor receptor (EGF receptor -2Erb B2) and epidermal growth factor receptor (EGF receptor -4e Erb B4) was detected by in situ Nick end labeling (Tunel) staining in order to detect cardiomyocyte apoptosis in rats with type 2 diabetic cardiomyopathy treated with NRG-1. Results the survival rate of cardiomyocytes increased from 63.33 卤3.56% to 85.88 卤4.55%, the content of ROS decreased from 33.57 卤4.23% to 15.88 卤4.55%, the apoptosis rate decreased from 36.44 卤4.86% to 14.77 卤4.21%, and the expression of Erb B2 and Erb B 4 receptors increased from 0.26 卤0.044 卤0.39 卤0.03 to 0.84 卤0.03 卤0.72 卤0.04, respectively. Compared with HG group, the activity of LDH and the content of MDA decreased gradually, but the activity of SOD increased gradually. Compared with the HG group, the apoptosis of cardiomyocytes in type 2 diabetic cardiomyopathy model rats with P0.05 and NRG-1 was significantly decreased. Conclusion: the effect of 1: NRG-1 on the survival rate of cardiomyocytes and the reduction of oxidative stress and apoptosis induced by high glucose may play a role by binding to Erb B2/Erb B4 receptor in cardiomyocytes.
【作者单位】：苏州市吴中人民医院心内科;苏州市吴中人民医院门诊部;成都军区昆明总医院地方干部病房科;
【分类号】：R587.2;R54

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