Down-regulation of ZnT8 expression in INS-1 rat pancreatic beta cells reduces insulin content and glucose-inducible insulin secretion.
Down-regulation of ZnT8 expression in INS-1 rat pancreatic beta cells reduces insulin content and glucose-inducible insulin secretion.
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DOI：
10.1371/journal.pone.0005679
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发表时间：
2009-05-25
期刊：
影响因子：
3.7
通讯作者：
Cohen DM
中科院分区：
文献类型：
--
作者：
Fu Y;Tian W;Pratt EB;Dirling LB;Shyng SL;Meshul CK;Cohen DM
SLC 30 A8基因编码胰腺β细胞表达的锌转运蛋白ZnT 8。SLC 30 A8基因的多态性与2型糖尿病的易感性相关，尽管这种表型的分子机制还不完全清楚。这种多态性可以通过影响基因产物的表达水平来发挥其作用。我们使用shRNA介导的方法在INS-1胰腺β细胞系中可重复地下调ZnT 8 mRNA表达>90%。ZnT 8下调的细胞表现出减少摄取外源性锌，确定使用锌敏感的报告染料，锌。ZnT 8-下调的细胞显示响应于高血糖刺激的降低的胰岛素含量和降低的胰岛素分泌（表示为总胰岛素含量的百分比），如通过胰岛素免疫测定所确定的。ZnT 8耗尽的细胞也表现出更少的致密核心囊泡通过电子显微镜。这些数据表明，在培养的胰腺β细胞中ZnT 8表达的降低引起对高血糖症的胰岛素应答的降低。此外，虽然我们没有提供直接的证据，这些数据表明，SLC 30 A8表达水平的多态性可能会影响胰岛素分泌和体内血糖反应。
The SLC30A8 gene codes for a pancreatic beta-cell-expressed zinc transporter, ZnT8. A polymorphism in the SLC30A8 gene is associated with susceptibility to type 2 diabetes, although the molecular mechanism through which this phenotype is manifest is incompletely understood. Such polymorphisms may exert their effect via impacting expression level of the gene product. We used an shRNA-mediated approach to reproducibly downregulate ZnT8 mRNA expression by >90% in the INS-1 pancreatic beta cell line. The ZnT8-downregulated cells exhibited diminished uptake of exogenous zinc, as determined using the zinc-sensitive reporter dye, zinquin. ZnT8-downregulated cells showed reduced insulin content and decreased insulin secretion (expressed as percent of total insulin content) in response to hyperglycemic stimulus, as determined by insulin immunoassay. ZnT8-depleted cells also showed fewer dense-core vesicles via electron microscopy. These data indicate that reduced ZnT8 expression in cultured pancreatic beta cells gives rise to a reduced insulin response to hyperglycemia. In addition, although we provide no direct evidence, these data suggest that an SLC30A8 expression-level polymorphism could affect insulin secretion and the glycemic response in vivo.
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