中国脑血管病杂志2012，Vol.9Issue(4)：200-205,6.DOI:10.3969/j.issn.1672-5921.2012.04.006
组织激肽释放酶对酸敏感离子通道1a介导的大鼠酸中毒神经元氧化应激的影响
Effect of tissue kallikrein on neuronal oxidative stress in acid-sensing ion channel 1a mediated acidosis in rats
摘要
Abstract
Objective To investigate the protective effect of tissue kallikrein (TK) on acidosia induced neuronal injury and the effect of TK on acid-sensing ion channel la (ASIC1a) mediated oxidative stress reaction. Methods Primary cultured 8-10 day cortical neurons of newborn SO rate were randomly assigned to normal control, acidosis (cells incubated in pH 6. 0 extracellular fluid for 4 hours), TK( 100 nmol/L), B2R agonist(bradykinin 100 nmol/L), ASICla blocker (PcTX, 100 ng/mL), and B2R antagonist (HOE140) pretreatment groups. Before giving the acid liquid treatment, the corresponding drug pretreatmenJ was given for 30 min in the TK, B2R agonist and ASICla hlocker pretreatment groups. At 30 min before TK intervention, B2R antagonist HOE140 (500 nmol/L) was administered, and acid liquid treatment was conducted after 30 min. A Cell counting Kit-8 (CCK-8) was used to detect the survival rate of neurons in each group. Different fluorescent probes were used to label intracellular reactive oxygen species ( ROS), nitric oxide ( NO), mitochondrial membrane potential ( MMP), and intracellular free Ca2+. A fluorescence microplale reader was used to delect the relative intensity of the fluorescent substances in cells in the above groups. The relative contents of various substances were calculated. Results ①The survival rates of neurons in the normal control, TK, ASIC la blocker, and B2R agonist groups were 96.6 ± 2.6% , 79.7 ± 5.9%, 74.2 ± 4.6% , and 77.7 ± 5. 0% , respectively. Compared to the acidosis group(59.0±6.0%), there were significant differences (P<0.01). The survival rate of neurons in the B2R antagonist group was 64.6 ± 3.8%. Compared to the TK group, there were significant differences (P < 0.01). ② Compared to the normal control group, the intracellular ROS, NO and free Ca2* levels were significantly higher (P<0.01), and the MMP level decreased significantly in the acidosis group (P<0.01); compared to the acidosis group, the intracellular ROS, NO, and Ca2+ levels decreased significantly, and the MMP level increased significantly in the TK, ASIC1a antagonist, and B2R antagonist groups (P<0.01 or P<0.05); and compared to the TK group, the intracellular ROS, NO and free Ca2+ levels increased significantly, and the MMP level decreased significantly in the B2R antagonist group (P<0.01). Conclusion In an acid environment, ASIC1 a activation mediates the neuronal oxidalive stress reaction and induces neuronal damage. TK may attenuate ASIC1a mediated oxidative stress injury via B2R activation.关键词
组织激肽释放酶/酸中毒/氧化性应激/神经元/酸敏感离子通道/大鼠Key words
Tissue kallikrein/ Acidosis/ Oxidative stress/ Neuron/ Acid-sensing ion channels/ Rats引用本文复制引用
刘玲,杨昉,刘海波,李敏,刘亚红,刘新峰,张仁良,董强..组织激肽释放酶对酸敏感离子通道1a介导的大鼠酸中毒神经元氧化应激的影响[J].中国脑血管病杂志,2012,9(4):200-205,6.基金项目
国家自然科学基金资助项目(81070923,81100870) （81070923,81100870）
