槲皮素通过NF-κB信号通路调控银屑病角质形成细胞的增殖、炎症和氧化应激OACSTPCD
Quercetin regulates proliferation,inflammation,and oxidative stress in psoriatic keratinocytes via the NF-κB signaling pathway
目的:槲皮素在银屑病治疗中具有良好的潜力,然而有关其具体的作用机制仍不清楚.本研究探讨槲皮素是否可通过核转录因子-κB(nuclear transcription factor-κB,NF-κB)信号通路调控银屑病角质形成细胞的增殖、炎症和氧化应激.方法:培养人表皮角质形成细胞(human epidermal keratinocytes,HaCaT),通过肿瘤坏死因子α(tumor necrosis factor α,TNF-α)诱导,建立银屑病细胞损伤模型(TNF-α组);然后再采用低、中、高剂量(浓度从低到高分别为0,3.75、7.50、15.00、30.00、60.00、120.00 μmol/L)槲皮素进行处理,并将其分为TNF-α+槲皮素低、中、高剂量组,将未进行任何处理的HaCaT细胞设为对照组.在TNF-α+槲皮素高剂量组细胞中,分别添加二甲基亚砜(dimethyl sulfoxide,DMSO)(TNF-α+槲皮素高剂量+DMSO组)和NF-κΒ通路激活剂(NF-κΒ activator 1)(TNF-α+槲皮素高剂量+NF-κΒ activator 1组)进行处理.采用细胞计数实验(cell counting kit-8,CCK-8)筛选槲皮素影响TNF-α刺激的细胞增殖的最小浓度;采用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)、生物化学染色和蛋白质印迹法分别检测HaCaT细胞炎症因子、氧化应激指标以及NF-κB信号通路相关蛋白的表达.结果:与TNF-α+槲皮素0 μmol/L细胞相比,TNF-α+槲皮素3.75、7.50、15.00、30.00 μmol/L细胞的增殖差异均无明显统计学意义(均P>0.05),而TNF-α+槲皮素60.00、120.00 μmol/L细胞的增殖受到显著抑制(均P<0.05).与对照组相比,TNF-α组细胞上清液中白细胞介素(interleukin,IL)-6、IL-1β、细胞间黏附分子-1(intercellular cell adhesion molecule-1,ICAM-1)和丙二醛(malondialdehyde,MDA)、磷酸化p65(phosphorylation-p65,p-p65)/p65的水平均显著升高(均P<0.001),而谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性显著下降(P<0.001).与TNF-α组相比,TNF-α+槲皮素中剂量组和TNF-α+槲皮素高剂量组细胞上清液中IL-6、IL-1β、ICAM-1和MDA的浓度均显著下降(均P<0.001),而GSH-Px活性均显著增加(均P<0.001),TNF-α+槲皮素高剂量组细胞中p-p65/p65水平显著降低(P<0.001).与TNF-α+槲皮素高剂量+DMSO组相比,TNF-α+槲皮素高剂量+NF-κΒ activator 1组细胞中p-p65/p65水平明显升高,细胞上清液中IL-6、IL-1β、ICAM-1和MDA的浓度均显著升高(P<0.001),而GSH-Px活性显著下降(P<0.001).结论:槲皮素通过抑制NF-κB信号通路调控银屑病角质形成细胞的增殖、炎症和氧化应激.
Objective:Quercetin holds promising potential in the treatment of psoriasis,yet its specific mechanisms of action remain unclear.This study investigates whether quercetin can regulate the proliferation,inflammation,and oxidative stress of psoriatic keratinocytes via the nuclear transcription factor kappa B(NF-κB)signaling pathway. Methods:Human epidermal keratinocytes(HaCaT)were cultured and induced with tumor necrosis factor α(TNF-α)to establish a psoriasis cell damage model(TNF-α group).The cells were then treated with low,medium,and high doses of quercetin(concentrations of 0,3.75,7.50,15.00,30.00,60.00,120.00 μmol/L),and categorized into TNF-α+quercetin low,medium,and high dose groups,with untreated HaCaT cells serving as the control group.In the TNF-α+high-dose quercetin group,cells were further treated with dimethyl sulfoxide(DMSO)(TNF-α+high-dose quercetin+DMSO group)and NF-κB pathway activator(NF-κB activator 1)(TNF-α+high-dose quercetin+NF-κB activator 1 group).The cell counting kit-8(CCK-8)assay was used to determine the minimal concentration of quercetin affecting TNF-α-induced cell proliferation.Enzyme-linked immunosorbent assay(ELISA),biochemical staining,and Western blotting were employed to detect inflammatory cytokines,oxidative stress markers,and NF-κB signaling pathway-related proteins in HaCaT cells. Results:Compared to TNF-α+quercetin 0 μmol/L cells,the proliferation of TNF-α+quercetin 3.75,7.50,15.00,and 30.00 μmol/L cells showed no significant differences(all P>0.05),while TNF-α+quercetin 60.00 and 120.00 μmol/L cells exhibited significantly inhibited proliferation(both P<0.05).Compared to the control group,the TNF-α group showed significantly elevated levels of interleukin(IL)-6,IL-1β,intercellular cell adhesion molecule-1(ICAM-1),malondialdehyde(MDA),and phosphorylation-65(p-p65)/p65(all P<0.001),and a significantly decreased level of glutathione peroxidase(GSH-Px)activity(P<0.001).Compared to the TNF-α group,the TNF-α+medium-dose quercetin and TNF-α+high-dose quercetin groups showed significantly reduced levels of IL-6,IL-1β,ICAM-1,and MDA in cell supernatants(all P<0.001),and significantly increased level of GSH-Px activity(P<0.001),with a notable reduction in p-p65/p65 levels in the TNF-α+high-dose quercetin group(P<0.001).Compared to the TNF-α+high-dose quercetin+DMSO group,the TNF-α+high-dose quercetin+NF-κΒ activator 1 group showed significantly elevated p-p65/p65 levels and increased concentrations of IL-6,IL-1β,ICAM-1,and MDA in cell supernatants(all P<0.001),and significantly decreased GSH-Px activity(P<0.001). Conclusion:Quercetin regulates the proliferation,inflammation,and oxidative stress in psoriatic keratinocytes by inhibiting the NF-κB signaling pathway.
朱宇;徐文静;张元瑜;段晓诚
湖南中医药大学第一附属医院皮肤科,长沙 410007
银屑病槲皮素增殖炎症氧化应激
psoriasisquercetinproliferationinflammationoxidative stress
《临床与病理杂志》 2024 (005)
647-655 / 9
湖南省自然科学基金(2023JJ60490);湖南省中医药科研计划项目(E2022004);长沙市自然科学基金(KQ2202457);湖南中医药大学校级科研基金(2022YYZK005);湖南中医药大学校级研究生创新课题基金(2022CX149).This work was supported by the Natural Science Foundation of Hunan Province(2023JJ60490),the Scientific Research Project of Traditional Chinese Medicine in Hunan Province(E2022004),the Changsha Natural Science Foundation(KQ2202457),the University Level Scientific Research Fund of Hunan University of Chinese Medicine(2022YYZK005),and the University Level Graduate Innovation Fund of Hunan University of Chinese Medicine(2022CX149),China.
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