摘要
目的:探讨钙释放通道稳定蛋白FKBP12.6过度表达对心力衰竭心室肌细胞肌浆网(SR)功能的影响。方法:用携带FKBP12.6基因的重组腺病毒Ad.FKBP12.6-GFP感染分离的心力衰竭心室肌细胞,通过RT-PCR和Western blotting技术检测转基因的表达;通过局部场刺激诱发胞内钙瞬变,SR钙容量则由咖啡因诱发的钙释放估测;以X-rhod-1-AM作为Ca2+指示剂,采用激光共聚焦线扫描检测细胞内Ca2+浓度的变化。结果:Ad-FKBP12.6-GFP组的FKBP12.6mRNA和蛋白表达水平分别比对照组升高5倍和4倍;Ad-FKBP12.6-GFP感染细胞的钙瞬变幅度(F/F0峰值,3.16±0.42vs1.43±0.38,P<0.01)和SR钙容量(F/F0峰值,4.15±0.54vs2.23±0.44,P<0.01)均显著高于Ad-GFP感染细胞。结论:采用基因转移技术上调FKBP12.6的表达可能是心力衰竭治疗的一条新途径。
AIM： To study the effects of FKS06 binding protein 12. 6 （ FKBP12. 6） overexpression on the performance of sarcoplasmic reticulum （SR） in ventricular myocytes of rats with heart failure. METHODS： The adenovirus （Ad） -mediated gene transfer was used to overexpress FKBP12. 6 in ventricular myocytes of rats with heart failure. Western blotting and reverse transcriptasepolymerase chain reaction （RT- PCR） analysis were used to reveale specific overexpression of FKBP12.6. X - rhod - 1 - AM was used as the Ca^2＋ indicator, and cells were viewed under a confocal microscope. RESULTS： Adenovirus mediated overexpression of FKBP12. 6 resulted in a 5 -fold increase in relative FKBP12. 6 mRNA levels and a 4 - fold increase in relative FKBP12. 6 protein levels at 48 h after transfection compared with control. The amplitude of the fluorescence [ Ca^2＋]i transient was significantly increased in Ad - FKBP12. 6 - GFP cardiomyocytes compared with Ad - GFP myocytes （ peak F/F0, 3.16±0.42 vs 1.43±0. 38, P 〈0.0 ）. The amplitude of the caffeine - evoked fluorescence [ Ca^2＋] i transient was also increased in FKBP12. 6 overexpressing myocytes compared with control myocytes （ peak F/F0, 4. 15± 0. 54 vs 2. 23 0. 44, P 〈0. 0 ）. CONCLUSION ： FKBP12. 6 overexpression by gene transfer technique improves SR performance.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2007年第7期1263-1266,共4页
Chinese Journal of Pathophysiology
基金
中国博士后科学基金资助项目(No2005037611)
