Andrographis paniculata and Its Bioactive Diterpenoids Against Inflammation and Oxidative Stress in Keratinocytes - PubMed (original) (raw)

Andrographis paniculata and Its Bioactive Diterpenoids Against Inflammation and Oxidative Stress in Keratinocytes

Eugenie Mussard et al. Antioxidants (Basel). 2020.

Abstract

Andrographis paniculata was widely used in traditional herbal medicine to treat various diseases. This study explored the potential anti-aging activity of Andrographis paniculata in cutaneous cells. Human, adult, low calcium, high temperature (HaCaT) cells were treated with methanolic extract (ME), andrographolide (ANDRO), neoandrographolide (NEO), 14-deoxyandrographolide (14DAP) and 14-deoxy-11,12-didehydroandrographolide (14DAP11-12). Oxidative stress and inflammation were induced by hydrogen peroxide and lipopolysaccharide/TNF-α, respectively. Reactive oxygen species (ROS) production was measured by fluorescence using a 2',7'-dichlorofluorescein diacetate (DCFH-DA) probe and cytokines were quantified by ELISA for interleukin-8 (IL-8) or reverse transcription-quantitative polymerase chain reaction (RT-qPCR) for tumor necrosis factor-α (TNF-α). Hyaluronic acid (HA) secretion was determined by an ELISA. Our results show a decrease in ROS production and TNF-α expression by ME (5 µg/mL) in HaCaT under pro-oxidant and pro-inflammatory conditions, respectively. ME protected HaCaT against oxidative stress and inflammation. Our findings confirm that ME can be used for the development of bioactive compounds against epidermal damage.

Keywords: Andrographis paniculata; andrographolide; inflammation; keratinocytes; oxidative stress; skin aging.

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Conflict of interest statement

To the best of our knowledge, no conflict of interest, financial or other, exists.

Figures

Figure 1

Figure 1

HPLC results of ANDRO from ME of Andrographis paniculata (a); ANDRO standard (b). Abbreviations: ANDRO, andrographolide; ME, methanolic extract.

Figure 2

Figure 2

Effect of ME, ANDRO, NEO, 14DAP, and 14DAP11-12 on HaCaT cytotoxicity. HaCaT was treated with increasing concentrations (1, 2.5, or 5 µg/mL) of ME (a,f), ANDRO (b,g), NEO (c,h), 14DAP (d,i), or 14DAP11-12 (e,j) for 24 h and 48 h. The control cells were treated with 0.05% DMSO (noted as “0”). Cell viability was analyzed by mitochondrial metabolism using an MTT assay (a–e). Then, cell cytotoxicity was determined by a dosage of extracellular LDH activity (f–j). Cells untreated with stimuli were a negative control and cells treated with lysis agent were a positive control (noted as “Control”). The values are mean ± SD, ** p < 0.01 compared with control group, n = 3. Abbreviations: NEO, neoandrographolide; 14DAP, 14-deoxyandrographolide; 14DAP11-12, 14-deoxy-11,12-didehydroandrographolide; DMSO, dimethyl sulfoxide; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; LDH, lactate dehydrogenase.

Figure 3

Figure 3

ROS production and the effect of ME, ANDRO, NEO, 14DAP, and 14DAP11-12 in HaCaT. ME (a), ANDRO (b), NEO (c), 14DAP (d), or 14DAP11-12 (e) was used at 1, 2.5, or 5 µg/mL for 1 h. The control cells were treated with 0.05% DMSO (noted as “0”). ROS production was induced by 0.5 mM H2O2 and free radical scavenging activity was done using a DCFH-DA probe. The values are mean ± SD, * p < 0.05 compared with control group, n = 3. Abbreviations: ROS, reactive oxygen species; DCFH-DA, 2′,7′-dichlorofluorescein diacetate.

Figure 4

Figure 4

TNF-α expression and effect of ME, ANDRO, NEO, 14DAP, and 14DAP11-12 in HaCaT under pro-inflammation conditions. ME (a), ANDRO (b), NEO (c), 14DAP (d), or 14DAP11-12 (e) was used at 1, 2.5, or 5 µg/mL for 24 h. The control cells were treated with 0.05% DMSO (noted as “0”). Inflammation conditions were induced by LPS (10 µg/mL) for 6 h and TNF-α expression was determined by RT-qPCR. The values are mean ± SD, * p < 0.05 compared with control group, n = 3. Abbreviations: LPS, lipopolysaccharides; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; TNF-α, tumor necrosis factor-α.

Figure 5

Figure 5

IL-8 secretion and effect of ME, ANDRO, NEO, 14DAP, and 14DAP11-12 in HaCaT under pro-inflammation conditions. ME (a), ANDRO (b), NEO (c), 14DAP (d), or 14DAP11-12 (e) was used at 1, 2.5, or 5 µg/mL for 24 h. The control cells were treated with 0.05% DMSO (noted as “0”). Inflammation was induced by TNF-α (10 ng/mL) and IL-8 secretion was performed using an ELISA assay. The values are mean ± SD, compared with control group, n = 3. Abbreviations: IL-8, interleukin-8.

Figure 6

Figure 6

HA production and effect of ME, ANDRO, NEO, 14DAP, and 14DAP11-12 in HaCaT. ME (a), ANDRO (b), NEO (c) 14DAP (d), or 14DAP11-12 (e) was used at 1, 2.5 or 5 µg/mL for 48h. HA was determined by an ELISA. The values are mean ± SD, compared with control group, n = 3. Abbreviations: HA, hyaluronic acid.

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References

    1. Cavinato M., Jansen-Dürr P. Molecular mechanisms of UVB-induced senescence of dermal fibroblasts and its relevance for photoaging of the human skin. Exp. Gerontol. 2017;94:78–82. doi: 10.1016/j.exger.2017.01.009. - DOI - PubMed
    1. Farage M.A., Miller K.W., Elsner P., Maibach H.I. Intrinsic and extrinsic factors in skin ageing: A review. Int. J. Cosmet. Sci. 2008;30:87–95. doi: 10.1111/j.1468-2494.2007.00415.x. - DOI - PubMed
    1. Pillai S., Oresajo C., Hayward J. Ultraviolet radiation and skin aging: Roles of reactive oxygen species, inflammation and protease activation, and strategies for prevention of inflammation-induced matrix degradation—A review. Int. J. Cosmet. Sci. 2005;27:17–34. doi: 10.1111/j.1467-2494.2004.00241.x. - DOI - PubMed
    1. Rinnerthaler M., Bischof J., Streubel M.K., Trost A., Richter K. Oxidative Stress in Aging Human Skin. Biomolecules. 2015;5:545–589. doi: 10.3390/biom5020545. - DOI - PMC - PubMed
    1. Tobin D.J. Introduction to skin aging. J. Tissue Viability. 2017;26:37–46. doi: 10.1016/j.jtv.2016.03.002. - DOI - PubMed

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