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KMID : 0613820180280121406
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Journal of Life Science
2018 Volume.28 No. 12 p.1406 ~ p.1415
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Gene Expression Profiles and Antioxidant Effects of Houttuynia cordata Thunb Extract in Human Keratinocyte HaCaT Cells
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Kim Jung-Min
Bang In-Soek
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Abstract
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Based on the antioxidative effects in organic solvent fractions obtained from the main methanolic extract of Houttuynia cordata Thunb, the cytoprotective effects by oxidative-stress were here analyzed. Regarding the antioxidant activity of organic solvent fractions, the electron-donating ability of DPPH increased in a dose-dependent manner, and ED50 exhibited the highest concentration at 175 μg/ml in the Hc-EtOAc fraction. The cell viability of Hc-EtOAc fractions on H2O2-induced HaCaT cell death (IC50) increased in a concentration-dependent manner and a visible cell survival rate of 74% was observed at a concentration of 100 μg/ml. Meanwhile, the gene expression patterns in HaCaT cells treated with 100 μg/ml of the Hc-EtOAc fraction for 6 and 24 hr were identified with microarray analysis. The genes involved in signal transduction, cell division, antioxidant activity, and epithelial cell proliferation were found to be 2-fold up-regulated genes in HaCaT cells following the Hc-EtOAc fraction treatment. Especially, proinflammatory cytokines (IL1B, TNF, and IL6) were identified as involved in antioxidant activity based on the expression patterns of the HaCaT cells, and pathway analysis indicated that TLR4 might be considered an upstream regulator of these genes. In order to verify the activity of IL1B, TNF, and IL6, qRT-PCR showed that the expression increased more than 2 times in HaCaT cells treated with at least 100 μg/ml of the Hc-EtOAc fraction. The activity of the upstream regulator TLR4 protein was also increased by the Hc-EtOAc fraction. As a result, the antioxidative activity of the Hc-EtOAc fraction is predicted to pass from TLR4 through cytokines such as IL1B, TNF, and IL6.
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KEYWORD
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Antioxidant activity, gene expression profiles, Houttuynia cordata, HaCaT cells, proinflammatory cytokines
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