肉桂醛促进高糖条件下皮肤成纤维细胞迁移及机制研究

目的 探讨肉桂醛(cinnamic aldehyde,CA),核因子E2相关因子(nuclear factor erythroid2-related factor,NRF2)通路激动剂对高糖条件下成纤维细胞迁移的影响及可能机制.方法 体外培养HS27细胞,分为正常对照组(NG,5.5 mmol/L)、高糖组(HG,30 mmol/L)、高糖(30 mmol/L)+肉桂醛(20μmol/L)组、高糖(30 mmol/L)+Con siRNA组、高糖(30 mmol/L)+NRF2 siRNA组.迁移实验观察CA对高糖条件下HS27迁移的影响.采用western blot分析CA干预后NRF2及下游HO-1,增殖和凋亡相关蛋白P21和P53的表达情况.2',7'-二氯荧光素二醋酸(DCF)法检测CA对高糖条件下细胞内活性氧簇(reactive oxygen species,ROS)水平的影响.采用NRF2siRNA抑制NRF2通路,观察HS27细胞迁移和氧化应激的变化.结果 与正常对照组相比,HS27在高糖情况下,迁移能力明显降低(P＜0.05),CA干预后,高糖对HS27迁移能力的抑制作用得到改善(P＜0.05).进一步研究发现,CA上调NRF2和HO-1,减轻高糖导致的ROS增加((4 360 935 ±1 445 723)vs(1 982 673 ±208 109),P＜0.05]).然而,高糖条件下敲除NRF2后,细胞内ROS产生进一步增多,并加剧对成纤维细胞迁移能力的抑制(0.046 ±0.018)vs(0.318 ±0.032),P＜0.05].结论 CA促进高糖情况下成纤维细胞迁移、缓解氧化应激损伤,并有可能部分依赖于NRF2通路激活.

Cinnamic aldehyde improves fibroblasts migration under hyperglycaemic condition and its underlying mechanism

Objective To determine the effect of cinnamic aldehyde (CA),a nuclear factor erythroid 2-related factor (NRF2) pathway activator,on the migration of fibroblasts (HS27 cells) and investigate the potential mechanism.Methods HS27 cells were divided into 5 groups,and were treated by normal glucose (NG,5.5 mmol/L),high glucose (HG,30 mmol/L),high glucose and CA (20 μ mol/L),high glucose and Con siRNA,high glucose and NRF2 siRNA,respectively.Polydimethylsiloxane (PDMS) assay was used to determine the effect of CA on cell mobility under high glucose condition.Western blotting was employed to examine the effect of CA on the expression of NRF2 and its downstream molecule HO-1,and proliferation and apoptosis related P21 and P53.Intracellular reactive oxide species (ROS) were measured in HS27 cells treated with CA using 2',7'-dichlorofluorescein diacetate (DCF) assay.NRF2 siRNA were transfected into HS27 cells to further validate the role of NRF2 pathway in the regulation of cell migration and oxidative stress.Results Compared with the HS27 cells under normal glucose condition,the migration of the cells cultured in high-glucose medium were significantly inhibited (P ＜ 0.05).After treatment with CA,the cell mobility was significantly improved (P ＜0.05) and the expression levels of NRF2 and HO-1 were notably increased.Meanwhile,CA treatment significantly ameliorated hyperglycemia-induced ROS production in HS27 cells (4 360 935 ± 1 445 723 vs 1 982 673 ± 208 109,P ＜ 0.05).However,knocking down of NRF2 far more enhanced ROS levels,and aggravated inhibitory effects of CA on migration of HS27 cells under high glucose condition (0.046 ±0.018 vs 0.318 ± 0.032,P ＜ 0.05).Conclusion CA treatment promotes the migration of fibroblasts and alleviates the oxidative stress damage from high glucose culture,which might be partially due to the activation of NRF2 pathway.