Reducing-Autophagy Derived Mitochondrial Dysfunction during Resveratrol Promotes Fibroblast-Like Synovial Cell Apoptosis
Reducing-Autophagy Derived Mitochondrial Dysfunction during Resveratrol Promotes Fibroblast-Like Synovial Cell Apoptosis
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DOI：
10.1002/ar.23798
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发表时间：
2018-07-01
影响因子：
2
通讯作者：
Chen, Xiaoyu
中科院分区：
文献类型：
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作者：
Cao, Wei;Zhang, Junqiang;Chen, Xiaoyu
类风湿关节炎患者滑膜成纤维细胞样细胞（FLS）的生长不受正常控制，但在组织学上与肿瘤细胞增殖相似。我们前期的研究表明，白藜芦醇抑制FLS的增殖，促进FLS的凋亡。然而，白藜芦醇诱导FLS凋亡的分子机制尚未确定。在这里，我们表明，FLS细胞活力（5 μ M H2 O2预处理24小时后）表现出更好的增殖性能比其他浓度通过CCK-8测定。TUNEL法和Western blotting法检测细胞凋亡率随白藜芦醇浓度（0、40、80、160、320 μ M）的增加而增加。免疫荧光和western blot分析显示，随着白藜芦醇浓度的增加，自噬相关蛋白（LC 3A/B、ATG-5）的表达水平降低。我们还发现，白藜芦醇诱导FLS线粒体形态的变化。此外，线粒体功能检测表明，随着白藜芦醇浓度的增加，线粒体膜电位的损失，由JC-1试验检测。细胞内ATP的产生与白藜芦醇浓度呈正相关和负相关。同时，随着白藜芦醇浓度的增加，细胞内钙释放和钙内流逐渐减少。因此，我们提出白藜芦醇可以降低FLS中的自噬水平。自噬水平的降低可导致活性氧的积累，从而导致线粒体功能障碍，促进FLS凋亡。(C)2018 Wiley Periodicals，Inc.
In rheumatoid arthritis patients, the fibroblast-like synovial cells (FLS) growth is not controlled normally, but is similar to the tumor cells proliferation in histology. Our previous studies have shown that resveratrol inhibits the proliferation of FLS and promotes FLS apoptosis. However, the molecular mechanisms involved in resveratrol-induced FLS apoptosis have not been determined yet. Here, we showed that the FLS cell viability (following pretreatment with 5 mu M H2O2 for 24 hr) exhibited better proliferation performance than at other concentrations via the CCK-8 assay. The cell apoptotic rate increased with the increasing concentration of resveratrol (0, 40, 80, 160, 320 mu M), as detected by TdT-mediated dUTP nick-end labeling (TUNEL) staining and western blotting. Furthermore, the expression level of autophagy-related proteins (LC3A/B, ATG-5) decreased with the increased concentration of resveratrol, as determined by immunofluorescence and western blot analysis. We also showed that resveratrol induced FLS mitochondrial morphology change. Moreover, mitochondrial function detection showed that the mitochondrial membrane potential was lost with the increased concentration of resveratrol as examined by the JC-1 assay. The production of ATP in cells was positively and negatively correlated with the resveratrol concentration. Simultaneously, the intracellular calcium release and calcium influx decreased gradually with the increase in resveratrol concentration. Therefore, we proposed that resveratrol can reduce the level of autophagy in FLS. The decrease in the autophagy level can lead to the accumulation of reactive oxygen species, which may result in mitochondrial dysfunction and promotion of FLS apoptosis. (C) 2018 Wiley Periodicals, Inc.