Pancreatic β-cells expressing GLP-1 are resistant to the toxic effects of immunosuppressive drugs
Pancreatic β-cells expressing GLP-1 are resistant to the toxic effects of immunosuppressive drugs
复制标题
DOI：
10.1677/jme.1.01655
复制
发表时间：
2005-04-01
影响因子：
3.5
通讯作者：
Perfetti, R
中科院分区：
文献类型：
--
作者：
D'Amico, E;Hui, HX;Perfetti, R
胰岛细胞移植后常发生糖耐量异常。免疫抑制剂（ISD）的管理，必要的，以避免组织排斥反应，是部分负责高血糖症。研究转染胰高血糖素样肽-1（GLP-1）前体基因片段的小鼠胰岛素瘤（MIN6）细胞（RIP/GLP-1 MINIS细胞）是否对ISD给药产生的毒性具有抗性。将RIP/GLP-1 MIN6细胞以及亲本MINIS细胞暴露于ISD的混合物。采用放射免疫分析和蛋白质印迹法检测胰岛素分泌和胰岛素相关蛋白的表达。通过流式细胞仪分析定量细胞凋亡。最后，为了研究GLP-1的抗凋亡作用是否是其对胰岛素分泌的作用的函数，或者更确切地说，它是GLP-1的直接作用，将细胞与或不与二氮嗪或毒蜥外泌肽-9一起培养。GLP-1改善了暴露于ISD的细胞的功能活性和活力。RIP/GLP-1 MIN6细胞在ISD作用后胰岛素分泌功能得到保护。P细胞表达GLP-1减少了凋亡细胞的数量，并增加了抗凋亡蛋白Bcl-2的表达。GLP-1还降低了促凋亡标志物PARP-p85和Smac/Diablo的丰度。用二氮嗪处理细胞并没有消除用GLP-1转染的细胞的保护优势;相反，细胞暴露于exendin-9与细胞凋亡易感性恢复相关。本报告证明GLP-1能够保护β细胞功能并保护细胞免于凋亡性细胞死亡。
Glucose intolerance is often observed after pancreatic islet cell transplantation. The administration of immunosuppressive agents (ISD), necessary to avoid tissue rejection, is in part responsible for hyperglycemia. To investigate whether mouse insulinoma (MIN6) cells transfected with the glucagon like peptide-1 (GLP-1) fragment of the proglucagon gene (RIP/GLP-1 MINIS cells) are resistant to the toxicity derived from the administration of ISD. RIP/GLP-1 MIN6 cells, as well as parental MINIS cells, were exposed to a cocktail of ISD. The secretion of insulin and the expression of apoptosis-related proteins were investigated by RIA and western blot analysis. Cell apoptosis was quantified by FACS analysis. Finally, to study whether the antiapoptotic action of GLP-1 was a function of its effect on insulin secretion, or rather it was a direct effect of GLP-1, cells were cultured with or without diazoxide or exendin-9. GLP-1 improved the functional activity and the viability of cells exposed to ISD. The insulin secretion of RIP/GLP-1 MIN6 cells after exposure to ISD was preserved. The expression of GLP-1 by P-cells reduced the number of apoptotic cells and increased the expression of the antiapoptotic protein Bcl-2. GLP-1 also decreased the abundance of the proapoptotic markers PARP-p85 and Smac/Diablo. Treatment of cells with the diazoxide did not abolish the protective advantage that cells transfected with GLP-1 had; conversely the exposure of cells to exendin-9 was associated with a restored susceptibility to apoptosis. This report demonstrates that GLP-1 is capable of preserving beta-cell function and protecting cells from apoptotic cell death.