基因工程改造的人胰腺β细胞系中的钙信号传导
Calcium signaling in a genetically engineered human pancreatic β-cell line.
作者信息
Krishnan Kalaiselvan, Ma Zuheng, Björklund Anneli, Islam Md Shahidul
机构信息
From the *Department of Clinical Science and Education, Södersjukhuset, Karolinska Institutet, Stockholm; Departments of †Molecular Medicine and Surgery, and ‡Endocrinology, Metabolism, and Diabetes, Karolinska University Hospital Solna, Stockholm; and §Department of Internal Medicine, Uppsala University Hospital, Uppsala, Sweden.
出版信息
Pancreas. 2015 Jul;44(5):773-7. doi: 10.1097/MPA.0000000000000318.
OBJECTIVES
The use of primary human β-cells for studying Ca signaling is limited by the scarcity of human pancreatic islets. Rodent insulinoma cell lines are widely used, but it is difficult to extrapolate results obtained from rodent cells to human. Recently, a genetically engineered human β-cell line EndoC-BH1 has been developed. We have examined whether the EndoC-BH1 cells could be used as a model for studying Ca signaling in the β-cells.
METHODS
We used microscope-based fluorometry to measure cytoplasmic-free Ca concentration from fura-2-loaded single EndoC-BH1 cells cultured on glass cover slips. Ca responses to different agonists of insulin secretion were studied. Insulin secretion was measured by radioimmunoassay.
RESULTS
EndoC-BH1 cells secreted insulin in response to glucose in a dose-dependent manner, and the secretion was enhanced by GLP-1 (glucagon-like peptide 1). Glucose, potassium chloride, carbachol, L-arginine, and tolbutamide increased cytoplasmic-free Ca concentration in the EndoC-BH1 cells. We found that GLP-1 was essential for Ca response to glucose and tolbutamide.
CONCLUSIONS
We concluded that the EndoC-BH1 cells can be used as model cells to study Ca signaling and stimulus-secretion coupling in the human β-cells.
目的
用于研究钙信号传导的原代人β细胞因人类胰岛稀缺而受到限制。啮齿动物胰岛素瘤细胞系被广泛使用，但将从啮齿动物细胞获得的结果外推至人类却很困难。最近，一种基因工程化的人β细胞系EndoC-BH1已被开发出来。我们研究了EndoC-BH1细胞是否可作为研究β细胞中钙信号传导的模型。
方法
我们使用基于显微镜的荧光测定法，来测量在玻璃盖玻片上培养的单个负载fura-2的EndoC-BH1细胞的胞质游离钙浓度。研究了钙对不同胰岛素分泌激动剂的反应。通过放射免疫测定法测量胰岛素分泌。
结果
EndoC-BH1细胞对葡萄糖以剂量依赖性方式分泌胰岛素，且胰高血糖素样肽1（GLP-1）可增强该分泌。葡萄糖、氯化钾、卡巴胆碱、L-精氨酸和甲苯磺丁脲可增加EndoC-BH1细胞中的胞质游离钙浓度。我们发现GLP-1对于钙对葡萄糖和甲苯磺丁脲的反应至关重要。
结论
我们得出结论，EndoC-BH1细胞可作为模型细胞，用于研究人β细胞中的钙信号传导和刺激-分泌偶联。
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