[논문]Oleanolic Acid Protects the Skin from Particulate Matter-Induced Aging

Kim, Youn Jin; Lee, Ji Eun; Jang, Hye Sung; Hong, Sung Yun; Lee, Jun Bae; Park, Seo Yeon; Hwang, Jae Sung

doi:10.4062/biomolther.2020.106

Oleanolic Acid Protects the Skin from Particulate Matter-Induced Aging 원문보기

Biomolecules & therapeutics, v.29 no.2, 2021년, pp.220 - 226

Kim, Youn Jin (Department of Genetic Engineering & Graduate School of Biotechnology, College of Life Sciences, Kyung Hee University) , Lee, Ji Eun (Department of Genetic Engineering & Graduate School of Biotechnology, College of Life Sciences, Kyung Hee University) , Jang, Hye Sung (Department of Genetic Engineering & Graduate School of Biotechnology, College of Life Sciences, Kyung Hee University) , Hong, Sung Yun (COSMAX R&I Center) , Lee, Jun Bae (COSMAX R&I Center) , Park, Seo Yeon (Creative & Innovation Center, IN2BIO) , Hwang, Jae Sung (Department of Genetic Engineering & Graduate School of Biotechnology, College of Life Sciences, Kyung Hee University)

Abstract ▼ AI-Helper

The role of particulate matter (PM) in health problems including cardiovascular diseases (CVD) and pneumonia is becoming increasingly clear. Polycyclic aromatic hydrocarbons, major components of PM, bind to aryl hydrocarbon receptor (AhRs) and promote the expression of CYP1A1 through the AhR pathway in keratinocytes. Activation of AhRs in skin cells is associated with cell differentiation in keratinocytes and inflammation, resulting in dermatological lesions. Oleanolic acid, a natural component of L. lucidum, also has anti-inflammation, anticancer, and antioxidant characteristics. Previously, we found that PM10 induced the AhR signaling pathway and autophagy process in keratinocytes. Here, we investigated the effects of oleanolic acid on PM10-induced skin aging. We observed that oleanolic acid inhibits PM10-induced CYP1A1 and decreases the increase of tumor necrosis factor-alpha and interleukin 6 induced by PM10. A supernatant derived from keratinocytes cotreated with oleanolic acid and PM10 inhibited the release of matrix metalloproteinase 1 in dermal fibroblasts. Also, the AhR-mediated autophagy disruption was recovered by oleanolic acid. Thus, oleanolic acid may be a potential treatment for addressing PM10-induced skin aging.

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표/그림 (7)

그림 Fig. 1. High-performance liquid chromatogram-PDA analysis. (A) Ligustrum lucidum ethanol extract (LL), (B) oleanolic acid and (C) L. lucidum ethyl acetate extract (LL-EA).
그림 Fig. 2. Effects of LL, LL-EA and oleanolic acid on keratinocyte viability. (A) HaCaT was treated with LL for 24 h. (B) HaCaT was treated with LL-EA and oleanolic acid for 24 h. Cell viability was assessed using the Ez-Cytox assay and measured at 450 nm. Values are presented as the mean ± SD of three determinations (n=3).
그림 Fig. 3. Inhibition of PM₁₀-induced CYP1A1 by LL, LL-EA and oleanolic acid in keratinocytes. (A) HaCaT was pre-treated with different concentrations of LL 20 μg/mL for 6 h, and after that PM₁₀ (50 μg/mL) was treated for 3 h and analyzed by RT-PCR. (B) HaCaT was pre-treated with different concentrations of LL-EA (2.5, 5, 10 μg/mL) for 6 h, and after that PM₁₀ (50 μg/mL) was treated for 3 h and analyzed by RTPCR. (C) HaCaT was pre-treated with different concentrations of oleanolic acid (2.5, 5, 10 μg/mL) for 6 h, and and after that PM₁₀ (50 μg/ mL) was treated for 3 h and analyzed by RT-PCR. Histograms of quantitated mRNA expression were showed in (D-F).
그림 Fig. 4. Inhibition of PM₁₀-induced TNF-α by LL-EA and oleanolic acid in keratinocytes. (A) HaCaT was pre-treated with different concentrations of LL-EA (2.5, 5, 10 μg/mL) or oleanolic acid (2.5, 5, 10 μg/mL) for 6 h, and after that PM₁₀ (50 μg/mL) was treated for 18 h and analyzed by RT-PCR. Histograms of quantitated mRNA expression were showed in (B).
그림 Fig. 5. Inhibition of PM₁₀-induced IL-6 by LL-EA and oleanolic acid in keratinocytes. HaCaT was pre-treated with different concentrations of LL-EA (2.5, 5, 10 μg/mL) or oleanolic acid (2.5, 5, 10 μg/ mL) for 6 h, and after that PM₁₀ (50 μg/mL) was treated for 24 h. Supernatants were used to analyze Enzyme linked immunosorbent assay (ELISA). Values are presented as the mean ± SD of three determinations (n=3). *p<0.05, **p<0.01 versus PM10 treatment group; ^##p<0.01 versus control group.
그림 Fig. 6. Regulation of MMP-1 in dermal fibroblast by supernatant derived from keratinocytes. HaCaT was pre-treated with different concentrations of oleanolic acid (2.5, 5, 10 μg/mL) for 24 h, and co-treated with oleanolic acid and PM₁₀ (100 μg/mL) for 30 min. After treatment, fresh media were changed and incubated for 24 h. HDFs were treated with the supernatant derived from HaCaT for 30 min. After treatment, fresh media were changed and incubated for 48 h. Supernatants derived from HDFs were used to analyze Enzyme linked immunosorbent assay (ELISA). Values are presented as the mean ± SD of three determinations (n=3). The MMP- 1 level of control was 93 ng/mL. **p<0.01 versus PM₁₀ treatment group; ^##p<0.01 versus control group.
그림 Fig. 7. Oleanolic acid decrease PM₁₀-induced autophagy process in keratinocyte. HaCaT were pre-treated with different concentrations of oleanolic acid (2.5, 5, 10 μg/mL) for 6h, and after that PM₁₀ (100 μg/mL) was treated for 24 h and analyzed by Western blot. β-actin was used as a loading control.

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제안 방법

HPLC-DAD analyses were carried out using an Agilent 1260 Inifinity (Agilent, Santa Clara, CA, USA). HPLC columns were silica-based C₁₈ Agilent Zorbax Elclipse Plus (250 mm×4.
lucidum on PM₁₀-induced skin aging are still unknown. Thus, in this study, we investigated the effects of oleanolic acid and two types of L. lucidum extract on PM₁₀-induced skin aging.

대상 데이터

Fine Dust (PM₁₀-like) (ERM-EZ100) was purchased from the European Reference Materials (ERM) (St. Louis, MO, USA). Fine Dust particles are composed of the size range of 10 μm (x ≤10 μm).
Human dermal fibroblasts cell line (HDF) was purchased from the American Type Culture Collection (ATCC) (VA, USA). Cells were cultured in DMEM (Welgene, Gyeongsan, Korea) medium supplemented with 10% Fetal Bovine serum (FBS), and 1% Penicillin/Streptomycin.
Human immortalized keratinocyte cell line (HaCaT) was obtained from Amore Pacific Company (Yongin, Korea). Human dermal fibroblasts cell line (HDF) was purchased from the American Type Culture Collection (ATCC) (VA, USA).
Fine Dust particles are composed of the size range of 10 μm (x ≤10 μm). α-naphthoflavone (α-NF) which is known as an inhibitor of AhR and used for positive control, and oleanolic acid were purchased from Sigma Aldrich (St. Louis, MO, USA).

데이터처리

Statistical significance was determined by independent t-test. A value of *p<0.

이론/모형

After treatment, fresh media were changed and incubated for 48 h. Supernatants derived from HDFs were used to analyze Enzyme linked immunosorbent assay (ELISA). Values are presented as the mean ± SD of three determinations (n=3).

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