摘要
目的观察小鼠心肌成纤维细胞(CFs)在心肌梗死后瞬时受体电位通道亚族M7(TRPM7)样电流的变化及其对胶原生成的影响,探讨TRPM7离子通道在心脏纤维化形成中的潜在病理生理作用。方法(1)制备小鼠心肌梗死模型并分离CFs细胞;(2)培养传代CFs及小分子干扰RNA(siRNA)技术感染;(3)应用膜片钳技术观察CFs缺血后TRPM7通道内外向电流特征;(4)钙荧光显像技术观察缺血心肌对CFs的钙离子内流影响;(5)测定缺血对CFs总胶原含量的影响。结果(1)心肌缺血能使CFs的含钙离子内向电流较对照组显著增加,分别为(7.4±0.7)pA/pF和(16.2±1.7)pA/pF(P〈0.05);(2)SiRNA使TRPM7样电流的mRNA水平明显下降,且电流幅值减少约90%;(3)心肌缺血组CFs的TRPM的mRNA丰度及总胶原含量较基础值增加2.3倍左右。结论致纤维化因子心肌缺血可通过TRPM7样电流介导的钙离子信号机制调节CFs功能,在心肌组织纤维化的病理生理过程中发挥重要作用。
Objective To investigate the changes of TRPM 7-like current in mouse cardiac fibroblast (CFs) after myocardial infarction and the effect of myocardial ischemia on the TRPM 7 expression and current. Methods (1) The model of myocardial infarction was made and CFs were isolated ; (2) CFs were cultured and infected by TRPM 7 siRNA ; (3) The effects of myocardial ischemia on TRPM 7 current were recorded by whole cell patch clamp technique; (4) The influences of myocardial ischemia on Ca^2+ influx in CFs were recorded by Ca^2+ fluorescence imaging. (5) The effects of ischemia on total collagen content of CFs were studied. Results (1) Ca^2+ inward current of CFs was increased after myocardial infarction ((16.2±1, 7) vs. (7.4±0.7) pA/pF, P〈0. 053 ; (2) TRPM 7 current was reduced by 90% after siRNA infeetion; (3) The total collagen content of CFs after ischemia was approximately 2.3-fold higher than basic value. Conclusions Ca^2+ influx in CFs plays an important role in the pathophysiological process of myocardial fibrosis.
出处
《中华老年医学杂志》
CAS
CSCD
北大核心
2008年第12期927-930,共4页
Chinese Journal of Geriatrics