论文部分内容阅读
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。
Objective To investigate the mechanism of action of berberine hydrochloride on keratinocytes. Methods The fluorescence intensity of Fluo-3/AM and Rhodamine-123 in a single living cell was observed before and after berberine hydrochloride treatment by laser scanning confocal microscopy and its technique. It was presumed that berberine hydrochloride could affect HaCaT cells. The effect of intracellular free calcium ion and mitochondrial membrane potential and the effect of energy metabolism. The results showed that Fluo-3/AM and Rhodamine-123-labeled HaCaT cells were observed to show that the fluorescence intensity of Fluo-3/AM was weak at rest and localized within the cells; the fluorescence intensity of Rhodamine-123 was The highest cell periphery, the lowest fluorescence intensity in the nuclear region. After PBS was added, the fluorescence intensity of Fluo-3/AM and rhodamine-123 in the cells did not change significantly. Berberine hydrochloride concentration-dependently enhanced the fluorescence intensity of Fluo-3/AM in HaCaT cells, but rapidly decreased the fluorescence intensity of Rhodamine-123. Conclusion Berberine hydrochloride may reduce the mitochondrial membrane potential of HaCaT cells by increasing the concentration of intracellular free calcium in HaCaT cells, which can inhibit the proliferation of keratinocytes.