Rattanaporn Patchara, Tongsima Sissades, Mandrup-Poulsen Thomas, Svasti Saovaros, Tanyong Dalina
Department of Clinical Microscopic, Faculty of Medical Technology, Mahidol University, Nakhon Pathom, Thailand.
Thalassemia Research Center, Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom, Thailand.
PeerJ. 2020 Jun 16;8:e9298. doi: 10.7717/peerj.9298. eCollection 2020.
Diabetes mellitus (DM) is a common complication found in β-thalassemia patients. The mechanism of DM in β-thalassemia patients is still unclear, but it could be from an iron overload and increase of some cytokines, such as interleukin1-β (IL-1β) and tumor necrosis factor-α (TNF-α). The objective of this study was to study the effect of interaction between ferric ammonium citrate (FAC) and cytokines, IL-1β and TNF-α, on 1.1B4 human pancreatic β-cell line.
The effect of the combination of FAC and cytokines on cell viability was studied by MTT assay. Insulin secretion was assessed by the enzyme-linked immunosorbent assay (ELISA). The reactive oxygen species (ROS) and cell apoptosis in normal and high glucose condition were determined by flow cytometer. In addition, gene expression of apoptosis, antioxidant; glutathione peroxidase 1 () and superoxide dismutase 2 (), and insulin secretory function were studied by real-time polymerase chain reaction (Real-time PCR).
The findings revealed that FAC exposure resulted in the decrease of cell viability and insulin-release, and the induction of ROS and apoptosis in pancreatic cells. Interestingly, a combination of FAC and cytokines had an additive effect on antioxidants' genes expression and endoplasmic reticulum (ER) stress. In addition, it reduced the insulin secretion genes expression; insulin (), glucose kinase (), protein convertase 1 (), and protein convertase 2 (). Moreover, the highest ROS and the lowest insulin secretion were found in FAC combined with IL-1β and TNF-α in the high-glucose condition of human pancreatic beta cell, which could be involved in the mechanism of DM development in β-thalassemia patients.
糖尿病（DM）是β地中海贫血患者中常见的并发症。β地中海贫血患者发生糖尿病的机制尚不清楚，但可能源于铁过载以及某些细胞因子如白细胞介素1-β（IL-1β）和肿瘤坏死因子-α（TNF-α）的增加。本研究的目的是研究柠檬酸铁铵（FAC）与细胞因子IL-1β和TNF-α之间的相互作用对1.1B4人胰腺β细胞系的影响。
采用MTT法研究FAC与细胞因子联合作用对细胞活力的影响。通过酶联免疫吸附测定（ELISA）评估胰岛素分泌。采用流式细胞仪测定正常和高糖条件下的活性氧（ROS）和细胞凋亡情况。此外，通过实时聚合酶链反应（Real-time PCR）研究凋亡、抗氧化剂、谷胱甘肽过氧化物酶1（）和超氧化物歧化酶2（）的基因表达以及胰岛素分泌功能。
研究结果显示，FAC暴露导致胰腺细胞活力和胰岛素释放降低，并诱导ROS和细胞凋亡。有趣的是，FAC与细胞因子的组合对抗氧化基因表达和内质网（ER）应激具有累加效应。此外，它降低了胰岛素分泌基因的表达，如胰岛素（）、葡萄糖激酶（）、蛋白转化酶1（）和蛋白转化酶2（）。此外，在人胰腺β细胞高糖条件下，FAC与IL-1β和TNF-α联合时发现ROS水平最高，胰岛素分泌最低，这可能与β地中海贫血患者糖尿病发生机制有关。
