枸橼酸铁铵对原代培养的大鼠心肌细胞铜蓝蛋白、膜铁转运辅助蛋白和膜铁转运蛋白表达的影响

目的 观察枸橼酸铁铵(FAC)对原代培养的大鼠心肌细胞铁释放相关蛋白表达的影响,探讨枸橼酸铁铵对心肌细胞铁代谢的影响机制. 方法 以原代培养的乳鼠心肌细胞为材料,分为对照组、20mg/L枸橼酸铁铵组、40mg/L枸橼酸铁铵组和80mg/L枸橼酸铁铵组,每组6个重复.然后检测心肌细胞存活率、搏动频率,免疫组织化学检测铜蓝蛋白(CP)、膜铁转运辅助蛋白(HP)和膜铁转运蛋白(FP1)表达的变化. 结果 各剂量枸橼酸铁铵对大鼠心肌细胞存活率无明显影响;心肌细胞搏动频率减慢,停止跳动的细胞数量明显增加,收缩幅度逐渐降低;随着枸橼酸铁铵浓度的增加,心肌细胞铜蓝蛋白、膜铁转运辅助蛋白和膜铁转运蛋白的表达均增加. 结论 枸橼酸铁铵影响大鼠心肌细胞的生理功能,IRP-IRE可能参与膜铁转运蛋白表达的调控,铜蓝蛋白、膜铁转运辅助蛋白表达的升高可能与铁处理增加细胞的氧化紧张性有关.

Effects of ferric ammonium citrate on the expression of proteins related to iron release in primary cultured rat cardiac myocytes

Objective To observe the effect of ferric ammonium citrate (FAC) on the pulsatile frequency and the expression of proteins related to iron releas in primary cultured rat cardiac myocytes. Methods Newborn rat cardiac myocytes were isolated and cultured with the medium containing FAC. The cells were divided into control group, 20mg/L FAC group, 40mg/L FAC group and 80mg/L FAC group. Every group had 6 repeats. The cells’ livability, beat rhythmically and beat amplitude of cardiac myocytes were detected. The expressions of ceruloplasmin (CP), hephaestin(HP) and ferroportion 1(FP1) were also examined by immunohistochemistry. Results Compared with control, FAC had no clearly influence on cells’ livability, but it could decrease cells’ beat rhythmically and beat amplitude. CP, HP and FP1 expression increased after the incubation of FAC. Conclusion FAC has no influence on cardiac myocytes’ livability, but it could affect the cells’ living state. IRP-IRE may be involved in the regulation of FP1 expression, the expression