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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。
Objective To investigate the effects of cinnamic aldehyde (CA) and nuclear factor erythroid 2-related factor (NRF2) pathway on the migration of fibroblasts under high glucose conditions and its possible mechanism. Methods HS27 cells were cultured in vitro and divided into normal control group (NG, 5.5 mmol / L), high glucose group (HG, 30 mmol / L), high glucose (30 mmol / L) and cinnamaldehyde (20μmol / High glucose (30 mmol / L) + Con siRNA group, high glucose (30 mmol / L) + NRF2 siRNA group. Effect of CA on HS27 Migration under High Glucose Conditions. Western blot was used to analyze the expressions of NR2 and downstream HO-1, proliferation and apoptosis-related proteins P21 and P53 after CA intervention. The effect of CA on the level of reactive oxygen species (ROS) in cells under high glucose condition was examined by 2 ’, 7’-dichlorofluorescein diacetate (DCF). NRF2 siRNA was used to inhibit the NRF2 pathway, and the changes of HS27 cell migration and oxidative stress were observed. Results Compared with the normal control group, HS27 had a significantly lower migration ability (P <0.05) under high glucose conditions. The inhibition of high glucose on HS27 migration was improved after CA intervention (P <0.05). Further studies found that CA up-regulates NRF2 and HO-1, and reduces the increase of ROS induced by high glucose ([4 360 935 ± 1 445 723] vs (982673 ± 208 109), P <0.05]. However, knockdown of NRF2 under high glucose condition further increased intracellular ROS production and aggravated the inhibition of fibroblast migration [(0.046 ± 0.018) vs (0.318 ± 0.032), P <0.05]. Conclusion CA can promote the migration of fibroblasts under high glucose condition and relieve the oxidative stress injury, and may partly depend on the activation of NRF2 pathway.