| 环孢霉素A对心肌细胞结构和胞内钙离子浓度的影响 |
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| 引用本文： | 王桂香,唐洁冰,傅羽,刘越,尹新华. 环孢霉素A对心肌细胞结构和胞内钙离子浓度的影响[J]. 心脏杂志, 2010, 22(4): 482-485 |
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| 作者姓名： | 王桂香 唐洁冰 傅羽 刘越 尹新华 |
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| 作者单位： | 哈尔滨医科大学附属第一临床医学院心内科，黑龙江 哈尔滨 150001 |
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| 基金项目： | 国家自然科学基金资助（30872387）；国家教育部博士基金资助（200802260006） |
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| 摘 要： | 目的:探讨不同浓度的环孢霉素A(cyclosporine,CsA)在不同时间点对心肌细胞形态及胞内钙离子浓度([Ca2+]i)的影响。方法:分别将1×10-3g/L、1×10-2g/L、2×10-2g/L的CsA与原代培养的Wistar乳鼠(1~3 d)心肌细胞(n=6)共孵育,于2 h、4 h、6 h、8 h和12 h后,采用相差显微镜观察各组心肌细胞的结构及收缩性;应用激光共聚焦显微镜观察心肌细胞内[Ca2+]i的变化。结果:不同浓度的CsA作用2 h后,与对照组相比较,各组心肌细胞的形态及收缩性无明显变化。1×10-3g/L和1×10-2g/L CsA组心肌细胞内的[Ca2+]i与对照组比较无明显差异;但2×10-2g/L CsA组心肌细胞内的[Ca2+]i较其他3组明显增加(P0.01)。作用4 h后,随着CsA浓度的增加,CsA组细胞的收缩性逐渐减弱,胞浆内可见小颗粒逐渐增多;各CsA组心肌细胞内的[Ca2+]i与对照组比较显著增加(P0.05);与1×10-3g/L CsA和对照组相比较,1×10-2g/L和2×10-2g/L CsA组增加明显(P0.05)。作用6 h后,随着CsA浓度的增加,各浓度CsA组与对照组比较,可见细胞内小空泡逐渐增多,细胞的收缩性逐渐减弱。各组心肌细胞内的[Ca2+]i随着CsA浓度的增加逐渐升高,各组组间具有显著差异(P0.05)。作用8 h和12 h后,1×10-3g/L CsA组的心肌细胞出现部分溶解,1×10-2g/L和2×10-2g/L CsA组心肌细胞的损伤严重,出现大面积死亡、溶解。结论:CsA可引起原代培养的心肌细胞损伤,收缩性减弱,且具有剂量和时间依赖性,可能与其引起的细胞内[Ca2+]i的增加有关。
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| 关 键 词： | 环孢霉素A 心肌细胞 细胞形态 钙离子浓度 大鼠 |
| 收稿时间： | 2010-02-21 |
Effect of cyclosporine A on cellular structure and intracellular calcium concentration in neonatal rat cardiomyocytes |
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| Abstract: | AIM: To evaluate the effect of cyclosporine (CsA) on the cellularity and intracellular calcium concentration ［Ca2+］i in neonatal rat cardiomyocytes. METHODS: Neonatal rat cardiomyocytes were prepared from 1- to 3-day-old neonatal Wistar rats. After CsA (1×10-3 g/L, 1×10-2 g/L, 2×10-2 g/L) was gastrically instilled into cardiomyocytes culture media randomly for 2, 4, 6, 8 and 12 h, the cellular structure and contractility were observed under phase-contrast microscope. ［Ca2+］i was measured by a laser scanning confocal microscope with excitation at 488 nm and emission at 530 nm. RESULTS: Compared with control, no obvious difference was observed in the cellularity of cardiomyocytes after incubation at different concentrations of the CsA for 2 h. ［Ca2+］i of cardiomyocytes significantly increased after incubation with CsA 2×10-2 g/L for 2 h compared with that in the other three groups (P<0.01). When the cardiomyocytes were incubated with CsA for 4 h, contractility of the cardiomyocytes was attenuated gradually and more particles were seen. ［Ca2+］i of the cardiomyocytes treated with each concentration of CsA increased compared with control (P<0.05). ［Ca2+］i of the cardiomyocytes treated with CsA 1×10-2 g/L and 2×10-2 g/L increased more significantly compared with that of cardiomyocytes treated with CsA 1×10-3 g/L. With treatment of CsA for 6 h, ［Ca2+］i of cardiomyocytes increased markedly (P<0.05) and more small vacuoles were seen in the cardiomyocytes. With treatment of CsA for 8 and 12 h and the increasing concentration of the CsA, more cardiomyocytes died. CONCLUSION: CsA impairs cardiomyocytes and attenuates contractility of cardiomyocytes in a concentration- and time-dependent manner of CsA. Impairment of the cardiomyocytes may be related to the effect that increased ［Ca2+］i of cardiomyocytes. |
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