3,3'-二吲哚甲烷对过氧化氢诱导人角质形成细胞氧化应激的预防作用及其机制

doi:10.3969/j.issn.1004-616x.2015.02.001

癌变·畸变·突变 ›› 2015, Vol. 27 ›› Issue (2): 81-85,90.doi: 10.3969/j.issn.1004-616x.2015.02.001

• 论著 • 下一篇

3,3'-二吲哚甲烷对过氧化氢诱导人角质形成细胞氧化应激的预防作用及其机制

殷花^1,2, 李文学², 李梦承³, 邓宇婷¹, 李金慧^1,2, 李军涛², 吴锦银², 杨光宇², 林忠宁⁴, 朱伟²

1. 中山大学公共卫生学院, 广东广州 510080;
2. 广州市疾病预防控制中心, 广东广州 510440;
3. 南方医科大学公共卫生与热带医学学院, 广东广州 510515;
4. 厦门大学公共卫生学院, 福建厦门 361102

收稿日期:2014-12-10 修回日期:2015-01-26 出版日期:2015-03-31 发布日期:2015-03-31
通讯作者: 朱伟,Tel:13710083695;E-mail:13710083695@126.com E-mail:13710083695@126.com
作者简介:殷花,E-mail:yinhua1989@126.com。
基金资助:
国家自然科学基金青年基金(81101562);广东省科技计划项目(2012B060300005);广东省自然科学基金(S2012010009633)

Preventive effect and mechanisms of 3,3'-diindolylmethane on oxidative stress induced by hydrogen peroxide in HaCaT cells

YIN Hua^1,2, LI Wenxue², LI Mengcheng³, DENG Yuting¹, LI Jinhui^1,2, LI Juntao², WU Jinyin², YANG Guangyu², LIN Zhongning⁴, ZHU Wei²

1. Department of Preventive Medicine, School of Public Health, Sun Yat-sen University, Guangzhou 510080, Guangdong;
2. Guangzhou Center for Disease Control and Prevention, Guangzhou 510440, Guangdong;
3. Southern Medical University, School of Public Health and Tropical Medicine, Guangzhou 510515, Guangdong;
4. College of Public Health, Xiamen University, Xiamen 361102, Fujian, China

Received:2014-12-10 Revised:2015-01-26 Online:2015-03-31 Published:2015-03-31
About author:10.3969/j.issn.1004-616x.2015.02.001

摘要/Abstract

摘要：

目的:探究3,3'-二吲哚甲烷(DIM)对过氧化氢(H₂O₂)诱导人角质形成细胞(HaCaT)氧化应激作用的预防效应及可能机制。方法:体外培养HaCaT细胞,用H₂O₂建立氧化应激模型。采用CCK-8法检测不同浓度(1~20 μmol/L)DIM对HaCaT细胞生长的抑制作用;流式细胞术检测DIM作用前后细胞内活性氧自由基(ROS)含量的变化;Western blot检测不同浓度DIM(0、5、10 μmol/L)对HaCaT氧化应激相关蛋白核因子(NF-κB)和丝裂原活化蛋白激酶(MAPKs)磷酸化表达水平的影响。结果:成功建立了HaCaT氧化应激模型。CCK-8法研究结果显示1~10 μmol/L DIM对HaCaT细胞无明显毒性作用(P>0.05);流式细胞术检测结果表明10 μmol/L DIM预处理可有效预防由H₂O₂诱导的HaCaT内ROS产生(P<0.05);Western blot检测结果显示,随着DIM浓度的增加,HaCaT中p38-MAPK、JNK和NF-κB磷酸化蛋白表达水平均逐渐降低(P均<0.05)。结论:DIM预处理可减弱HaCaT中由H₂O₂引起的氧化应激,其机制可能是通过抑制ROS的产生及对NF-κB、MAPK家族等氧化应激相关蛋白表达的调控作用来实现的,提示DIM可作为预防或改善氧化应激型皮肤细胞损伤的一种有效药物。

关键词: 3,3'-二吲哚甲烷, HaCaT细胞, 过氧化氢, 氧化应激

Abstract:

OBJECTIVE:To observe the effects of 3, 3'-diindolylmethane (DIM) on oxidative stress induced by hydrogen peroxide (H₂O₂) in HaCaT cells. METHODS:HaCaT cells were treated with different concentrations of H₂O₂ to establish the models of oxidative stress. CCK-8 assay was performed to evaluate the inhibitive effects of DIM (1-20 μmol/L) on cells proliferation. The intracellular reactive oxygen species (ROS) levels were measured with flow cytometry. Furthermore, the effects of DIM (0, 5 and 10 μmol/L) on H₂O₂-induced the phosphorylation level of nuclear factor κB (p-NF-κB), the expression of mitogen activated protein kinase (MAPKs) were detected with Western blot. RESULTS: The cell model of oxidative stress was successfully established. The results of CCK-8 assay showed that in the doses of 1-10 μmol/L, DIM did not have obvious influence on cell viability (P>0.05). Flow cytometry results indicated that pre-treatment with DIM (10 μmol/L) could inhibit the level of intracellular ROS (P<0.05). With increasing concentration of DIM, the levels of p-p38-MAPK, p-JNK and p-NF-κB were significantly depressed. CONCLUSION: DIM could protect HaCaT cells from H₂O₂-induced oxidative stress via suppressing production of ROS levels and down-regulating the expression of NF-κB and members of MAPKs. DIM might be used as an effective drug to treat or reduce oxidative stress-mediated injury in skin cells.

Key words: 3,3'-diindolylmethane, HaCaT cells, hydrogen peroxide, oxidative stress

中图分类号:

R730.2

殷花, 李文学, 李梦承, 邓宇婷, 李金慧, 李军涛, 吴锦银, 杨光宇, 林忠宁, 朱伟. 3,3'-二吲哚甲烷对过氧化氢诱导人角质形成细胞氧化应激的预防作用及其机制[J]. 癌变·畸变·突变, 2015, 27(2): 81-85,90.

YIN Hua, LI Wenxue, LI Mengcheng, DENG Yuting, LI Jinhui, LI Juntao, WU Jinyin, YANG Guangyu, LIN Zhongning, ZHU Wei. Preventive effect and mechanisms of 3,3'-diindolylmethane on oxidative stress induced by hydrogen peroxide in HaCaT cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2015, 27(2): 81-85,90.

参考文献

[1] Abida O, Gargouri B, Ben Mansour R, et al. Biomarkers of oxidative stress in epidermis of Tunisian pemphigus foliaceus patients[J]. J Eur Acad Dermatol Venereol, 2013, 27(3): 271-275.
[2] Verma AK, Prasad SB. Changes in glutathione, oxidative stress and mitochondrial membrane potential in apoptosis involving the anticancer activity of cantharidin isolated from redheaded blister beetles, epicauta hirticornis[J]. Anticancer Agents Med Chem, 2013, 13(7):1096-1114.
[3] Ines D, Sonia B, Fatma BA, et al. Date seed oil inhibits hydrogen peroxide-induced oxidative stress in human epidermal keratinocytes[J]. Int J Dermatol, 2010, 49(3):262-268.
[4] Liu L, Xie H, Chen X, et al. Differential response of normal human epidermal keratinocytes and HaCaT cells to hydrogen peroxide-induced oxidative stress[J]. Clin Exp Dermatol, 2012, 37(7):772-780.
[5] Shim JH, Kim KH, Cho YS, et al. Protective effect of oxidative stress in HaCaT keratinocytes expressing E7 oncogene [J]. Amino Acids, 2008, 34(1):135-141.
[6] Banerjee S, Kong D, Wang Z, et al. Attenuation of multi-targeted proliferation-linked signaling by 3, 3'-diindolylmethane (DIM):from bench to clinic[J]. Mutat Res, 2011, 728(1/2): 47-66.
[7] Beaver LM, Yu TW, Sokolowski EI, et al. 3, 3'-Diindolylmethane, but not indole-3-carbinol, inhibits histone deacetylase activity in prostate cancer cells[J]. Toxicol Appl Pharmacol, 2012, 263(3):345-351.
[8] Luo H, Wang L, Schulte BA, et al. Resveratrol enhances ionizing radiation-induced premature senescence in lung cancer cells[J]. Int J Oncol, 2013, 43(6):1999-2006.
[9] Vayalil PK, Mittal A, Hara Y, et al. Green tea polyphenols prevent ultraviolet light-induced oxidative damage and matrix metalloproteinases expression in mouse skin[J]. J Invest Dermatol, 2004, 122(6):1480-1487.
[10] Rojkind M, Dominguez-Rosales JA, Nieto N, et al. Role of hydrogen peroxide and oxidative stress in healing responses[J]. Cell Mol Life Sci, 2002, 59(11):1872-1891.
[11] Shimamoto K, Hayashi H, Taniai E, et al. Antioxidant N-acetyl-L-cysteine(NAC) supplementation reduces reactive oxygen species (ROS)-mediated hepatocellular tumor promotion of indole-3-carbinol (I3C) in rats[J]. J Toxicol Sci, 2011, 36(6):775-786.
[12] Benabadji SH, Wen R, Zheng JB, et al. Anticarcinogenic and antioxidant activity of diindolylmethane derivatives[J]. Acta Pharmacol Sin, 2004, 25(5):666-671.
[13] Nho CW, Jeffery E. Crambene, a bioactive nitrile derived from glucosinolate hydrolysis, acts via the antioxidant response element to upregulate quinone reductase alone or synergistically with indole-3-carbinol[J]. Toxicol Appl Pharmacol, 2004, 198(1):40-48.
[14] Bowie A, O'neill LA. Oxidative stress and nuclear factor-kappaB activation:a reassessment of the evidence in the light of recent discoveries[J]. Biochem Pharmacol, 2000, 59(1):13-23.
[15] Dudek EJ, Shang F, Taylor A. H(2)O(2)-mediated oxidative stress activates NF-kappa B in lens epithelial cells[J]. Free Radic Biol Med, 2001, 31(5):651-658.
[16] Yao Z, Hu W, Yin S, et al. 3, 3'-Diindolymethane ameliorates adriamycin-induced cardiac fibrosis via activation of a BRCA1-dependent anti-oxidant pathway[J]. Pharmacol Res, 2013, 70(1):139-146.
[17] Fecher LA, Amaravadi RK, Flaherty KT. The MAPK pathway in melanoma[J]. Curr Opin Oncol, 2008, 20(2):183-189.
[18] Sakai N, Wada T, Furuichi K, et al. Involvement of extracellular signal-regulated kinase and p38 in human diabetic nephropathy[J]. Am J Kidney Dis, 2005, 45(1):54-65.
[19] Baeuerle PA. The inducible transcription activator NF-kappa B:regulation by distinct protein subunits[J]. Biochim Biophys Acta, 1991, 1072(1):63-80.

3,3'-二吲哚甲烷对过氧化氢诱导人角质形成细胞氧化应激的预防作用及其机制

Preventive effect and mechanisms of 3,3'-diindolylmethane on oxidative stress induced by hydrogen peroxide in HaCaT cells

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