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目的探讨茶多酚对长波紫外线(UVA)诱导HaCaT细胞急性光损伤的防护作用及其机制。方法用光学显微镜、MTT法检测UVA对细胞形态及增殖活性影响,构建UVA致HaCaT细胞急性光损伤模型;用MTT法检测茶多酚对细胞增殖活性影响,获得茶多酚对HaCaT细胞最适浓度;检测茶多酚处理前后细胞增殖活性(MTT法)、细胞膜损伤(LDH)情况;Western blot、RT-PCR检测茶多酚处理前后核因子NF-E2相关因子2(Nuclear factor erythroid 2-related factor 2,Nrf2)蛋白胞内分布、Nrf2 mRNA及下游Ⅱ相解毒酶表达情况。结果 20 J/cm~2 UVA致HaCaT细胞形态学改变且抑制细胞增殖活性(0.11±0.50;P<0.05),可用于构建UVA致HaCaT细胞急性光损伤模型;茶多酚剂量依赖性抑制细胞增殖活性,11μg/mL为本实验最适茶多酚浓度;相比对照组、UVA组的细胞增殖活性(1.043±0.203;0.113±0.050)和LDH(1.000±0.092;1.858±0.016),照射前加入茶多酚可显著提高细胞增殖活性(1.322±0.080;均P<0.05)和减少LDH释放(0.500±0.079;均P<0.05);Western blot、RT-PCR结果显示,茶多酚可显著增加核内Nrf2蛋白表达,上调Nrf2 mRNA(1.804±0.229)及下游Ⅱ相解毒酶SOD mRNA(1.172±0.148)、CAT mRNA(1.617±0.259)、GCLC mRNA(1.183±0.083)、GCLM mRNA(1.228±0.256)的表达。结论茶多酚可有效防护UVA诱导HaCaT细胞急性光损伤,且茶多酚可增加核内Nrf2蛋白及其mRNA和下游Ⅱ相解毒酶mRNA的表达,茶多酚的光防护作用可能与Nrf2信号通路有关。
Objective To investigate the protective effect and mechanism of tea polyphenols on acute photodamage induced by long-wave ultraviolet (UVA) in HaCaT cells. Methods The effect of UVA on cell morphology and proliferation activity was detected by MTT, and the model of acute photodamage caused by UVA was constructed. The effect of tea polyphenols on cell proliferation was tested by MTT assay. The optimal concentration of tea polyphenols for HaCaT cells (MTT) and cell membrane damage (LDH) were detected before and after the treatment of tea polyphenols. The levels of nuclear factor erythroid 2-related factor 2 2, Nrf2) intracellular distribution of protein, Nrf2 mRNA and phase Ⅱ downstream detoxification enzyme expression. Results The morphological changes and cell proliferation inhibition of HaCaT cells induced by 20 J / cm ~ 2 UVA (0.11 ± 0.50; P <0.05) could be used to establish the model of acute light injury induced by UVA. HaCaT cells inhibited the proliferation of HaCaT cells in a dose- Activity, 11μg / mL was the optimum concentration of tea polyphenols in this experiment. Cell proliferation activity (1.043 ± 0.203; 0.113 ± 0.050) and LDH (1.000 ± 0.092; 1.858 ± 0.016) in UVA group were Tea polyphenols significantly increased cell proliferation (1.322 ± 0.080; all P <0.05) and LDH release (0.500 ± 0.079; all P <0.05). The results of Western blot and RT-PCR showed that tea polyphenols could significantly increase the nuclear (1.172 ± 0.148), CAT mRNA (1.617 ± 0.259), GCLC mRNA (1.183 ± 0.083) and GCLM mRNA (1.228 ± 0.256) were significantly increased in Nrf2 mRNA and Nrf2 mRNA (1.804 ± 0.229 and 1.172 ± 0.148, expression. Conclusion Tea polyphenols can effectively protect the HaCaT cells from acute light damage induced by UVA, and tea polyphenols can increase the expression of Nrf2 protein and its mRNA and the phase Ⅱ detoxification enzyme mRNA in the nucleus. The photoprotection of tea polyphenols may be related to the Nrf2 signaling pathway related.