A multi-method evaluation of the effects of Inflammatory cytokines (IL-1 beta, IFN-gamma, TNF-alpha) on pancreatic beta-cells
A multi-method evaluation of the effects of Inflammatory cytokines (IL-1 beta, IFN-gamma, TNF-alpha) on pancreatic beta-cells
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多种方法评估炎症细胞因子（IL-1β、IFN-γ、TNF-α）对胰腺 β 细胞的影响
DOI：
10.1002/jcp.26518
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发表时间：
2018
影响因子：
5.6
通讯作者：
Tang Wei
中科院分区：
文献类型：
--
作者：
Xie Kaipeng;Xu Bo;Zhang Yuqing;Chen Minjian;Ji Yinwen;Wang Jie;Huang Zhenyao;Zhou Kun;Xia Yankai;Tang Wei
我们的目的是探讨炎症因子（IL - 1β, IFN - γ, TNF - α）对胰腺β -细胞的影响。CCK‐8实验显示，TNF‐α处理24小时，IFN‐γ处理48小时，IL‐1β处理84小时后，细胞活力下降。EdU -标记的红色荧光细胞在治疗24小时后，TNF - α组显著减少，而IFN - γ和IL - 1β组则无明显减少。流式细胞术结果显示，TNF - α和IFN - γ组细胞凋亡增加，而IL - 1β组细胞凋亡增加。细胞凋亡结果发现，IL - 1β和TNF - α组的S期细胞数量增加，而IFN - γ组与对照组的细胞周期无显著差异。TEM图像显示，IL - 1β和IFN - γ组的颗粒和线粒体数量减少，特别是TNF - α组的胰岛素颗粒和线粒体数量减少。放射免疫分析结果显示，TNF‐α抑制葡萄糖诱导的胰岛素分泌，而IL‐1β和IFN‐γ组与对照组相比无显著变化。代谢组学分析发现，氨基酸代谢和克雷布斯循环是炎症细胞因子治疗后最强大的代谢途径。总之，氨基酸代谢和克雷布斯循环代谢的改变可能是TNF - α诱导小鼠胰腺β细胞功能障碍的重要机制。
We aimed to explore the effects of Inflammatory cytokines (IL‐1β, IFN‐γ, TNF‐α) on pancreatic β‐cells. CCK‐8 assay showed that the cell viability decreased after 24 hr treatment of TNF‐α, 48 hr of IFN‐γ, and 84 hr of IL‐1β. EdU assay illustrated that after 24 hr treatment, there were significantly reduced EdU‐labeled red fluorescence cells in TNF‐α group while not in IFN‐γ and IL‐1β groups. Flow Cytometry results displayed that TNF‐α and IFN‐γ groups increased apoptosis while IL‐1β group did not. Cell apoptosis results found that there was an increase in the S‐phase population of IL‐1β and TNF‐α groups, however, there was no significant difference in cell cycle between IFN‐γ group and the control. TEM images showed that there were reduction in the number of granules and mitochondria in IL‐1β and IFN‐γ groups, in particular paucity of insulin granules and mitochondria in TNF‐α group. Radioimmunoassay results presented that TNF‐α inhibited glucose‐induced insulin secretion, while there were no significant changes in IL‐1β and IFN‐γ groups when compared with the control. Metabolomic analysis found amino acid metabolism and Krebs cycle were the most robust altered metabolism pathways after inflammatory cytokines treatments. Overall, the altered amino acid metabolism and Krebs cycle metabolism might be important mechanisms of TNF‐α induced mouse pancreatic β‐cells dysfuction.