中华皮肤科杂志 ›› 2005, Vol. 38 ›› Issue (2): 105-107.

• 论著 • 上一篇    下一篇

盐酸小檗碱对HaCaT细胞内游离钙离子及线粒体膜电位的影响

石继海1, 鞠强1, 尹兴平1, 辛燕1, 康晓静1, 张超英2, 夏隆庆1   

  1. 1. 中国医学科学院、中国协和医科大学皮肤病研究所 南京 210042;
    2. 南京师范大学 南京 210042
  • 收稿日期:2004-01-21 出版日期:2005-02-15 发布日期:2005-02-15
  • 通讯作者: 夏隆庆,E-mail:xialqing@public1.ptt.js.cn E-mail:xialqing@public1.ptt.js.cn

Berberine-induced Changes of Mitochondria Membrane Potential and Free Intracellular Calcium of HaCaT Cells

SHI Ji-hai1, JU Qiang1, YIN Xing-ping1, XIN Yan1, KANG Xiao-jing1, ZHANG Chao-ying2, XIA Long-qing1   

  1. Institute of Dermatology, Chinese Academy of Medical Sciences & Peking Union Medical College, Nanjing 210042, China
  • Received:2004-01-21 Online:2005-02-15 Published:2005-02-15

PDF

107

赞

摘要： 目的 探讨盐酸小檗碱对角质形成细胞的作用机制。方法 用激光扫描共聚焦显微镜及其技术观察盐酸小檗碱作用前后,单个活细胞内Fluo-3/AM及罗丹明-123(Rhodamine-123)荧光强度的变化,推测盐酸小檗碱对HaCaT细胞内游离钙离子和线粒体膜电位的作用及能量代谢的影响。结果 对Fluo-3/AM及罗丹明-123标记的HaCaT细胞观察发现:静息状态下,Fluo-3/AM的荧光强度较弱,细胞内呈局限性分布;罗丹明-123的荧光强度在细胞周边最高,核区的荧光强度最低。PBS加入后,细胞内Fluo-3/AM及罗丹明-123荧光强度没有明显变化。盐酸小檗碱呈浓度依赖性持续增强HaCaT细胞内Fluo-3/AM荧光强度,但迅速降低罗丹明-123荧光强度。结论 盐酸小檗碱可能通过增加HaCaT细胞内游离钙离子的浓度,降低HaCaT细胞线粒体膜电位,达到其抑制角质形成细胞增殖的作用。

关键词: 小檗碱, 细胞, 培养的, 阳离子, 二价, 线粒体, 膜电位

Abstract: Objective To study the effects on berberine hydrochloride on the mitochondria membrane potential and free intracellular calcium of HaCaT cells, and elucidate the mechanism of action of berberine on keratinocytes. Methods Rhodamine-123 fluorescence (very sensitive to mitochondria membrane potential) and Fluo-3/AM fluorescence (suitabe to detect free intracellular calcium in single HaCaT cell) were measured by laser scanning confocal technique. Results Fluo-3/AM fluorescence intensity of HaCaT cells was persistently increased after treating with berberine at concentrations of 5×10-5M, 2.5×10-5M and 1.25×10-5M, and significant differences were observed as compared with the PBS control. The intensity of rhodamine-123 fluorescence in HaCaT cells was decreased immediately when exposed to berberine, with significant difference from that of the PBS control. Conclusions It is suggested that berberine could increase free intracellular calcium and decrease mitochondria membrane potential of HaCaT cells, induce overload of intracellular calcium, influence energy metabolism, and then inhibit the proliferation of keratinocytes.

Key words: Berberine, Cells, cultured, Cations, divalent, Mitochondria, Membrane potentials