›› 2009, Vol. 8 ›› Issue (5): 256-259.

• 基础研究 • 上一篇    下一篇

钙三醇干预甲状旁腺素诱导的心肌细胞内钙超载和细胞肥大

赵卫红 张晓文 王笑云   

  1. 南京医科大学第一附属医院
  • 收稿日期:2008-10-27 修回日期:1900-01-01 出版日期:2009-05-12 发布日期:2009-05-12
  • 通讯作者: 赵卫红

Protective roles of calcitriol on overload of intracellular calcium and hypertrophy of cardiomyocytes induced by parathyroid hormone

ZHAO Wei-hong, ZHANG Xiao-wen, WANG Xiao-yun   

  1. The First Affiliated Hospital of Nanjing Medical University
  • Received:2008-10-27 Revised:1900-01-01 Online:2009-05-12 Published:2009-05-12

摘要:

【摘要】目的 探讨甲状旁腺素(PTH)对鼠心肌细胞内游离钙([Ca2+]i)和心肌肥大的影响及其机制,以及钙三醇的干预作用。方法 培养的新生大鼠心肌细胞以Fluo-3/AM负载,通过激光共聚焦显微术(LSCM)测定细胞内[Ca2+]i;以细胞面积和细胞蛋白含量作为心肌细胞肥大指标,体外实验观察PTH瞬时和持续刺激对鼠心肌细胞[Ca2+]i和肥大的影响,以及钙三醇的干预作用。结果 1.在细胞外Ca2+为2.5mmol/L时,PTH1-34在0.1和1μmol浓度下的刺激可促使心肌细胞静息[Ca2+]i荧光强度(FI)快速上升;1μmol浓度PTH1-34刺激,在细胞外液无钙,或应用10μmol硝苯地平预处理时,心肌细胞静息钙则无明显上升。2.培养的心肌细胞应用PTH1-34 0.01 和0.1μmol刺激7天后,心肌细胞内钙荧光强度、心肌细胞面积和蛋白含量均较对照组显著增加。若以PTH1-340.1μmol刺激,并同时加入0.001μmol的钙三醇干预,上述指标明显降低(P<0.01);但应用0.1μmol高浓度钙三醇干预,上述指标则未见显著改善。 结论 PTH1-34刺激可显著增加培养心肌细胞[Ca2+]i,诱导细胞肥大;适宜浓度钙三醇具有保护作用,电压依赖型钙通道的开放引起的细胞外钙内流增加是PTH1-34诱导上述变化的重要机制之一。

关键词: 钙三醇, 甲状旁腺素(PTH), 心肌细胞, 细胞内游离钙([Ca2+]i), 心肌肥大

Abstract:

【Abstract】 Objective To observe the effects of calcitriol on intracellular calcium concentration ([Ca2+]i) and hypertrophy of cardiomyocytes induced by parathyroid hormone (PTH). Materials and Methods Cultured neonatal rat cardiomyocytes were loaded with Fluo-3/AM, and [Ca2+]i was determined by a laser confocal microscope. Hypertrophic response of neonatal rat cardiomyocytes was assayed by measuring cell surface area and protein content. Results (a) When the extracellular Ca2+ was 2.5mmol/L, rapid stimulation of PTH1-34 at the concentration of 0.1 and 1μmol/L significantly elevated fluorescence intensity of [Ca2+]i in cardiomyocytes (P<0.01). However, when the extracellular fluid contained no calcium or the cardiomyocytes were pre-treated with 10μmol/L nifedipine, 1μmol/L PTH1-34 did not induce the increase of fluorescence intensity any more. (b) When the cardiomyocytes were treated in 0.01 and 0.1μmol/L PTH1-34 for 7 days, their surface area, protein content and [Ca2+]i increased significantly, especially those treated in 0.1μmol/L PTH1-34. If the cells were co-treated with 0.001μmol/L calcitriol and 0.1μmol.L-1 PTH1-34, their surface area, protein content and [Ca2+]i attenuated significantly. However, when the calcitriol concentration increased to 0.1μmol/L in the co-treatment experiment, these changes remained. Conclusion Cardiomyocyte hypertrophy was induced by PTH1-34, and was inhibited by appropriate concentration of calcitriol. Calcium influx through voltage-dependent channel induced by PTH may be one of the important mechanisms leading to cardiomyocyte hypertrophy.

Key words: Parathyroid hormone (PTH), Cardiomyocytes, Intracellular calcium concentration ([Ca2+]i), Hypertrophy