论文部分内容阅读
目的用消逝波显微镜观察胰岛β细胞瘤来源的Ins-1细胞的胰岛素出胞作用,旨在为“一吻即逝”胰岛素分泌方式提供新的证据。方法用含囊泡相关膜蛋白2和增强型绿色荧光蛋白融合蛋白基因的质粒标记Ins-1细胞的胰岛素分泌囊泡,用含高葡萄糖和高氯化钾的溶液刺激Ins-1细胞,用基于物镜的消逝波显微镜观察胰岛素的出胞作用。结果 Ins-1细胞经高葡萄糖高氯化钾溶液刺激后,消逝波显微镜可观察到两种现象:①荧光基团(亮点)在细胞膜出现并一直或长期停留在细胞膜;②荧光基团在细胞膜出现,停留较短时间后又逐渐消失(离开细胞膜)。前者应属于传统的膜融合分泌方式,即胰岛素分泌囊泡与细胞膜融合,其内容物被释放,囊泡膜成为细胞膜的一部分。后者应属于“一吻即逝”分泌方式,即胰岛素分泌囊泡与细胞膜接触并部分融合,释放部分内容物后,囊泡离开细胞膜,囊泡膜不成为细胞膜的一部分。结论该项研究结果支持胰岛素的分泌方式,除了传统的膜融合方式外,还存在“一吻即逝”的分泌方式。
Objective To observe the insulin exocytosis of Ins-1 cells derived from pancreatic islet cell tumors by evanescent wave microscope and to provide new evidence for the “kissing” insulin secretion. Methods Insulin-secreting vesicles of Ins-1 cells were labeled with plasmids containing vesicle-associated membrane protein 2 and enhanced green fluorescent protein fusion protein, Ins-1 cells were stimulated with a solution containing high glucose and potassium chloride, Evanescent objective lens observation of insulin exocytosis. Results After stimulated by high glucose and potassium chloride solution, there were two phenomena observed in the evanescent wave microscope: (1) the fluorescent group (bright spot) appeared in the cell membrane and remained in the cell membrane for a long time or in the long term; (2) Appear, stay for a short time and then gradually disappear (leave the cell membrane). The former should belong to the traditional way of membrane fusion, that is, insulin secretion vesicles and cell membrane fusion, the release of its contents, vesicular membrane as part of the cell membrane. The latter should belong to the “kiss kiss farewell” secretion, that is, insulin secretion vesicles contact with the cell membrane and part of the fusion, the release of part of the content, the vesicles leave the cell membrane, vesicle membrane does not become part of the cell membrane. Conclusion The findings support the insulin secretion, in addition to the traditional way of membrane fusion, there is a “kiss” of the secretion.