枸杞多糖对紫外线辐射HaCaT细胞急性损伤中Nrf2／Bach1通路的影响

目的 探讨转录因子Nrf2/Bach1在枸杞多糖（LBP）防御紫外线（UV）致HaCaT 细胞光损伤的作用。方法 体外培养的HaCaT细胞经300 μg/mL LBP预处理24 h后,分别接受30 J/cm2UVA及300 mJ/cm2UVB照射,孵育24h。以噻唑蓝（MTT法）检测细胞增殖活性; 尼罗红荧光染色法检测过氧化脂质含量; 采用单细胞凝胶电泳法检测DNA损伤; RT-qPCR法检测Nrf2和Bach1 mRNA及下游II相解毒酶基因的表达,Western blot法检测Nrf2及Bach1蛋白在细胞内分布及表达情况。结果 强度为30 J/cm2UVA及300 mJ/cm2UVB均可...

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Published in中国皮肤性病学杂志 Vol. 31; no. 11; pp. 1169 - 1174
Main Author 李振洁;彭丽倩;江娜;刘清;张尔婷;莫子茵;代歆悦;李华平;梁碧华;李润祥;朱慧兰
Format Journal Article
LanguageChinese
Published 2017
Subjects
Bach1
HaCaT细胞
Nrf2
多糖类
枸杞
紫外线
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Summary:目的 探讨转录因子Nrf2/Bach1在枸杞多糖（LBP）防御紫外线（UV）致HaCaT 细胞光损伤的作用。方法 体外培养的HaCaT细胞经300 μg/mL LBP预处理24 h后,分别接受30 J/cm2UVA及300 mJ/cm2UVB照射,孵育24h。以噻唑蓝（MTT法）检测细胞增殖活性; 尼罗红荧光染色法检测过氧化脂质含量; 采用单细胞凝胶电泳法检测DNA损伤; RT-qPCR法检测Nrf2和Bach1 mRNA及下游II相解毒酶基因的表达,Western blot法检测Nrf2及Bach1蛋白在细胞内分布及表达情况。结果 强度为30 J/cm2UVA及300 mJ/cm2UVB均可造成HaCaT细胞增殖能力下降,过氧化脂质含量及DNA荧光强度、迁移距离增加,与空白组相比,差异均有统计学意义（P〈0.05）; 300 μg/mL LBP预处理可显著提高细胞增殖活性,降低过氧化脂质水平,减轻DNA链断裂损伤,且增加Nrf2核蛋白及Bach1质蛋白的量,促进Nrf2 mRNA及下游II相解毒酶SOD,CAT,NQO1,GCLC,GCLM mRNA的表达,抑制Bach1mRNA的表达（均P〈0.05）。结论 枸杞多糖可有效减轻UV诱导的HaCaT细胞氧化损伤,其机制可能是通过激活Nrf2/Bach1转位及下游II相解毒酶基因的表达,发挥光防护作用。
Bibliography:LI Zhen-jie 1, 2, PENG Li-qian1, 2, 3, JIANG Na1, 2, LIU Qing1, 2, ZHANG Er-ting1, 2, 3, MO Zi-yin1, 2, 3, DAI Xin-yue1, 2, LI Hua-ping1, 2, LIANG Bi-hua1.2, LI Run-xiang1, 2, ZHU Hui-lan1, 2（1.Guangzhou Institute of Dermatology, Guangzhou 510095, China, 2.Institute of Dermatology Guangzhou Medical University, Guangzhou 510095, China, 3.Guangzhou Medical University, Guangzhou 511436, China）
Objective To evaluate the role of transcriptional factors of Nrf2 and Bach1 in the prevention of LBP against UV-induced damage in HaCaT cells.Methods The HaCaT cells cultured in vitro were pretreated with 300 μg/mL of LBP for 24 hours, and then were irradiated with 30 J/cm2UVA and 300 mJ/cm2UVB respectively.The cell viability was inspected by the MTT assay.Lipid peroxide levels were detected by the Nile red staining.Then the DNA strand breaks were evaluated through the comet assay.We employed real time PCR to detect the expressions of Nrf2 Bach1 mRNA, and the expressions of phase II enzyme genes.Western blot was carried out
ISSN:1001-7089