山奈酚减轻人角质形成细胞氧化应激损伤  

Kaempferol alleviates oxidative stress injury of human keratinocytes

作　　者：邹雪莲 胡雯[2,3] 雷子贤 王红娟 徐晨[2,3] 哈丽娜·海若拉 康晓静 ZOU Xue-lian;HU Wen;LEI Zi-xian;WANG Hong-juan;XU Chen;Halina HAIRUOLA;KANG Xiao-jing(Xinjiang Clinical College,Anhui Medical University,Urumqi 830001;Department of Dermatology,People's Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830001;Xinjiang Key Laboratory of Dermatology Research,Department of Science and Technology of Xinjiang Uygur Autonomous Region, Urumqi 830001, China)

机构地区：[1]安徽医科大学新疆临床学院,新疆乌鲁木齐830001 [2]新疆维吾尔自治区人民医院皮肤性病科,新疆乌鲁木齐830001 [3]新疆维吾尔自治区科技厅新疆皮肤病研究重点实验室,新疆乌鲁木齐830001

出　　处：《基础医学与临床》2021年第8期1103-1108,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(81760563)。

摘　　要：目的探究驱虫斑鸠菊成分山奈酚(KP)在人角质形成细胞系(HaCaT)中的抗氧化作用及作用机制。方法用选择性反应/多反应监测技术定量分析驱虫斑鸠菊种子黄酮类物质。将HaCaT细胞分为对照组、模型组[用2,2-偶氮二(2-甲基丙基咪)二盐酸盐(AAPH)处理HaCaT细胞,构建氧化应激模型]、不同浓度山奈酚干预组;CCK-8法检测增殖;RT-qPCR检测Nrf2、HO-1、NQO1、GCLC和GCLM mRNA表达;Western blot检测Nrf2蛋白表达。结果1)驱虫斑鸠菊种子中总黄酮类物质含量为0.97 g/100 g,共获得36种黄酮类化合物,其中KP含量为62.96 ng/100 g。2)构建氧化应激模型的AAPH最适浓度为25 mmol/L;KP浓度为0~40μmol/L时,未见明显细胞毒性。3)低剂量KP可增加HaCaT细胞内Nrf2、HO-1和GCLC mRNA的表达,其中血红素加氧酶-1(HO-1)的表达变化最为显著。4)KP可呈浓度依赖性增加各组细胞内Nrf2蛋白表达。结论驱虫斑鸠菊成分KP减轻人角质形成细胞氧化应激损伤,其作用机制与上调核因子E2相关因子2(Nrf2)的表达有关。Objective To explore the antioxidant effect and mechanism of kaempferol(KP),a component of Vernonia anthelmintica(L.)Willd.,in human immortalized keratinocytes(HaCaT).Methods Using selective reaction/multiple response monitoring technique for quantitative analysis of the flavonoids in the seeds of Vernonia anthelmintica(L.)Willd..HaCaT cells were divided into control group,model group(HaCaT cells were treated with AAPH to establish the oxidative stress model),low-,middle-and high-dose KP intervention groups.CCK-8 assay for proliferation,RT-qPCR was used for checking the expression of Nrf2,HO-1,NQO1,GCLC and GCLM mRNA,Western blot was used for checking the expression of Nrf2 protein.Results 1)The content of total flavonoids of Vernonia anthelmintica(L.)Willd..was 0.97 g/100 g,and a total of 36 flavonoid compounds were quantified,the content of KP was 62.96 ng/100 g.2)The optimum concentration of AAPH for constructingoxidative stress model was 25 mmol/L,and when the concentration of KP was 0-40μmol/L,no obvious cytotoxicity was found.3)Low-dose KP increased the expression of Nrf2,HO-1 and GCLC mRNA.Among them,the expression of HO-1 changed most significantly.4)KP increased the expression of Nrf2 protein in a concentration-dependent manner.Conclusions A component of Vernonia anthelmintica(L.)Willd.,reduces the oxidative stress injury of human keratinocytes,the mechanism might be related to the up-regulation of Nrf2 expression.

关 键 词：驱虫斑鸠菊 山奈酚 白癜风 氧化应激 角质形成细胞 

分 类 号：R758.4+1[医药卫生—皮肤病学与性病学]