吉林大学学报（医学版）2017，Vol.43Issue(5)：881-886,后插3,7.DOI:10.13481/j.1671-587x.20170505
N-乙酰半胱氨酸通路对AGT-REN双转基因高血压小鼠心肌成纤维细胞中电导钙激活钾离子通道蛋白表达的影响及其意义
Effect of N-acetyl cysteine pathway on expression of intermediate conductance Ca2+-activated K+ channels in cardiac fibroblasts of AGT-REN double transgenic hypertension mice
摘要
Abstract
Objective:To investigate the relationship between the level of oxidative stress and the expression of intermediate conductance calcium activated potassium channel (KCa3.1) protein in the cardiac fibroblasts (CFs) during hypertension process,and to clarify the role of KCa3.1 in cardiac fibrosis and its mechanism.Methods:The CFs of male C57B6 and AGT-REN double transgenic hypertension (dTH) mice were cultured and the wild C57B6 mouse CFs were used as control group.The CFs of dTH mice were randomly divided into high blood pressure group (dTH) and N-acetyl cysteine group (NAC).The CFs were treated with different concentrations of NAC for 24 h.The cell proliferation was detected by MTT and double dichlorofluorescein (DCFH-DA) probe was used for the detection of cellular reactive oxygen species (ROS) expression;Western blotting was employed to detect the expressions of collagen Ⅰ,collagen Ⅲ,Kca3.1 channel protein and the changes of PI3K signaling pathway protein phosphorylation.Results:The ROS production and protein expression of Kca3.1 channel of the dTH mice on 4,8,12 months were increased compared with 2 months (P＜0.05 or P＜0.01);the results of MTT suggested that the proliferation rates of CFs were 165.9％,138.72％,110.92％ and 109.82％ after administration of 1×10-6,1× 10 5,1× 10 4 and 1 × 10-3 mol · L-1 NAC in the dTH mice,and 1 × 10-4 and 1 × 10 3 mol · L-1 NAC significantly inhibited the proliferation of CFs.Compared with control group,the secretion of collagen Ⅰ and Ⅲ of CFs in the TH mice was decreased in 1 × 10-4 mol · L-1 NAC group (P＜0.01).The results of Western blotting showed that compared with control group,the expression level of Kca3.1 channel protein in CFs of the TH mice in 1 × 10-4 mol · L-1 NAC group was decreased (P＜0.01).Compared with control group,the p-AKT/T-AKt in CFs of the dTH mice was increased (P＜0.01);but in NAC group,the p-AKT/T-AKt was lower than that in dTH group (P＜0.01).Conclusion:NAC can inhibit the expression of KCa3.1 channel protein in CFs of the dTH hypertensive mice,which may be related to increasing the phosphorylation of AKt/PI3K signaling pathway.关键词
心肌纤维化/活性氧/心肌成纤维细胞/中电导钙激活钾离子通道Key words
myocardial fibrosis/reactive oxygen species/cardiac fibroblasts/intermediatec onductance Ca2+-activated K+ channels分类
医药卫生引用本文复制引用
王丽萍,王建辉,李树民,杨秀红..N-乙酰半胱氨酸通路对AGT-REN双转基因高血压小鼠心肌成纤维细胞中电导钙激活钾离子通道蛋白表达的影响及其意义[J].吉林大学学报（医学版）,2017,43(5):881-886,后插3,7.基金项目
国家自然科学基金资助课题(81600316,81372029) （81600316,81372029）
河北省科技厅自然科学基金资助课题(H2015209153) （H2015209153）
