山奈酚减轻人角质形成细胞氧化应激损伤

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (8): 1103-1108.

山奈酚减轻人角质形成细胞氧化应激损伤

邹雪莲^1,2, 胡雯^2,3, 雷子贤^2,3, 王红娟^2,3, 徐晨^2,3, 哈丽娜·海若拉^2,3, 康晓静^1,2*

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

收稿日期:2021-02-18 修回日期:2021-05-25 出版日期:2021-08-05 发布日期:2021-07-21
通讯作者: *drkangxj666@163.com
基金资助:
国家自然科学基金(81760563)

Kaempferol alleviates oxidative stress injury of human keratinocytes

ZOU Xue-lian^1,2, HU Wen^2,3, LEI Zi-xian^2,3, WANG Hong-juan^2,3, XU Chen^2,3, Halina HAIRUOLA^2,3, KANG Xiao-jing^1,2*

1. Xinjiang Clinical College,Anhui Medical University,Urumqi 830001;
2. Department of Dermatology,People's Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830001;
3. Xinjiang Key Laboratory of Dermatology Research,Department of Science and Technology of Xinjiang Uygur Autonomous Region, Urumqi 830001, China

Received:2021-02-18 Revised:2021-05-25 Online:2021-08-05 Published:2021-07-21
Contact: *drkangxj666@163.com

摘要/Abstract

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0～40 μmol/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。

关键词: 驱虫斑鸠菊, 山奈酚, 白癜风, 氧化应激, 角质形成细胞

Abstract: 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 constructing oxidative stress model was 25 mmol/L, and when the concentration of KP was 0-40 μmol/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.

Key words: Vernonia anthelmintica (L.) Willd., kaempferol, vitiligo, oxidative stress, keratinocytes

中图分类号:

R758.4+1

邹雪莲, 胡雯, 雷子贤, 王红娟, 徐晨, 哈丽娜·海若拉, 康晓静. 山奈酚减轻人角质形成细胞氧化应激损伤[J]. 基础医学与临床, 2021, 41(8): 1103-1108.

ZOU Xue-lian, HU Wen, LEI Zi-xian, WANG Hong-juan, XU Chen, Halina HAIRUOLA, KANG Xiao-jing. Kaempferol alleviates oxidative stress injury of human keratinocytes[J]. Basic & Clinical Medicine, 2021, 41(8): 1103-1108.

参考文献

[1]Cui T, Zhang W, Li S, et al. Oxidative stress-induced HMGB1 release from melanocytes: a paracrine mechanism underlying the cutaneous inflammation in vitiligo[J]. J Invest Dermatol, 2019, 139:2174-2184.
[2]Richmond JM, Bangari DS, Essien KI, et al. Keratinocyte-derived chemokines orchestrate T-Cell positioning in the epidermis during vitiligo and may serve as biomarkers of disease[J]. J Invest Dermatol, 2017, 137:350-358.
[3]Chen X, Guo W, Chang Y, et al. Oxidative stress-in-duced IL-15 trans-presentation in keratinocytes contributes to CD8⁺ T cells activation via JAK-STAT pathway in vitiligo[J]. Free Radic Biol Med, 2019, 139:80-91.
[4]Qiu L, Song Z, Setaluri V. Oxidative stress and vitiligo: the Nrf2-ARE signaling connection[J]. J Invest Dermatol, 2014, 134:2074-2076.
[5]石得仁, 普雄明, 沈大为, 等. 复方斑鸠菊擦剂治疗白癜风的疗效观察[J]. 新疆医学, 2005,35:49-50.
[6]Fouzder C, Mukhuty A, Kundu R. Kaempferol inhibits Nrf2 signalling pathway via downregulation of Nrf2 mRNA and induces apoptosis in NSCLC cells[J]. Arch Biochem Biophys, 2021, 697:108700.doi:10.1016/j.abb.2020.108700.
[7]Wang JY, Chen H, Wang YY, et al. Network pharmacolo-gical mechanisms of Vernonia anthelmintica (L.) in the treatment of vitiligo: isorhamnetin induction of melanogenesis via up-regulation of melanin-biosynthetic genes[J]. BMC Syst Biol, 2017, 11:103.doi:10.1186/s12918-017-0486-1.
[8]Barygina V, Becatti M, Lotti T, et al. ROS-challenged keratinocytes as a new model for oxidative stress-mediated skin diseases[J]. J Cell Biochem, 2019, 120:28-36.
[9]赵璐, 孙俊波, 金小琴. 丙二醛通过抑制Nrf2/ARE促进肾小球系膜细胞凋亡[J]. 基础医学与临床, 2015, 35:69-73.
[10]刘守珠, 王海源, 司立宁, 等. 血红素加氧酶-1减轻糖尿病小鼠心肌损伤[J]. 基础医学与临床, 2019, 39:170-174.
[11]Jian Z, Li K, Song P, et al. Impaired activation of the Nrf2-ARE signaling pathway undermines H₂O₂-induced oxidative stress response: a possible mechanism for melanocyte degeneration in vitiligo[J]. J Invest Dermatol, 2014, 134:2221-2230.
[12]Zhang S, Yi X, Su X, et al. Ginkgo biloba extract protects human melanocytes from H₂O₂-induced oxidative stress by activating Nrf2[J]. J Cell Mol Med, 2019, 23:5193-5199.
[13]Hu Y, Huang J, Li Y, et al. Cistanche deserticola polysaccharide induces melanogenesis in melanocytes and reduces oxidative stress via activating NRF2/HO-1 pathway[J]. J Cell Mol Med, 2020, 24:4023-4035.
[14]Yao H, Sun J, Wei J, et al. Kaempferol protects blood vessels from damage induced by oxidative stress and inflammation in association with the Nrf2/HO-1 signaling pathway[J]. Front Pharmacol, 2020, 11:1118.doi:10.3389/fphar.2020.01118.

[1]	赵肖灵, 何润之, 孙玉姣, 张小冲, 杨林芳, 王瑞, 程芳. 活动期白癜风患者血清γ干扰素含量升高[J]. 基础医学与临床, 2022, 42(7): 1067-1070.
[2]	郭婧, 李亚洁, 牟寒霜. LncRNA FGD5-AS1靶向miR-106a-5p减轻ox-LDL诱导的HUVECs损伤[J]. 基础医学与临床, 2022, 42(5): 768-775.
[3]	鲍敏, 林玫, 甄海宁, 何芳, 丁敏, 袁竹青, 陈亚隽, 薛欣欣. 醛糖还原酶抑制剂抑制屋尘螨诱导的人支气管上皮细胞系BEAS-2氧化应激反应[J]. 基础医学与临床, 2022, 42(4): 616-621.
[4]	李杰, 浦洋, 张梦迪, 张鹏举, 许寅喆. mTOR活化上调MGST1并增强细胞抗氧化能力[J]. 基础医学与临床, 2022, 42(4): 553-559.
[5]	张欣, 廖晓辉. 线粒体功能障碍在急性肾损伤向慢性肾脏疾病转变中的作用[J]. 基础医学与临床, 2021, 41(4): 589-592.
[6]	王海滨, 门庆玲, 蔡艳丽, 周文龙, 郭海志, 陈瑞祥. 左乙拉西坦对缺氧缺血性脑损伤新生大鼠的神经保护作用及氧化应激反应的影响[J]. 基础医学与临床, 2021, 41(4): 545-550.
[7]	李玉婷, 罗乐, 尹素娟, 朱小兰. 丹酚酸B减轻LPS诱导的小鼠急性肺损伤[J]. 基础医学与临床, 2021, 41(3): 376-381.
[8]	黄伟孔丽君叶亚群王宏伟. AMPK激活剂减轻重症急性胰腺炎模型大鼠胰腺损伤[J]. 基础医学与临床, 2021, 41(11): 1606-1611.
[9]	杨晓丽王征张艳茹冀静静王孟帅. 丁苯酞减轻H2O2诱导的人脐静脉和脐动脉内皮细胞氧化损伤[J]. 基础医学与临床, 2021, 41(10): 1451-1456.
[10]	谢飞, 周美兰. 羟基红花黄色素A抑制高糖诱导的小鼠肾足细胞损伤[J]. 基础医学与临床, 2021, 41(1): 27-32.
[11]	于锐, 张新茹, 王丹丹, 何平, 白瑜, 田密, 张蓓茹. 肾损伤分子-1参与碘对比剂致人肾小管上皮细胞体系HK-2凋亡[J]. 基础医学与临床, 2020, 40(9): 1206-1211.
[12]	胡苏琴, 王圣凯, 吴杭菲, 郑靖宇, 李剑敏. miR-195抑制剂降低1型糖尿病小鼠肝脏氧化应激损伤[J]. 基础医学与临床, 2020, 40(6): 811-816.
[13]	刘明, 杨臣礼, 孟庆鑫. 紫草素减轻低氧-复氧致大鼠心肌细胞系H9c2损伤[J]. 基础医学与临床, 2020, 40(3): 380-387.
[14]	黎笑兰, 张新广, 尹少萍. 白芨多糖抑制溃疡性结肠炎大鼠炎性反应与氧化应激[J]. 基础医学与临床, 2020, 40(2): 224-228.
[15]	孔婷婷刘明慧王琴琴姜云璐王春梅. 食欲素-A降低大鼠脑缺血-再灌注损伤[J]. 基础医学与临床, 2019, 39(9): 1239-1242.