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Studies on Physiological Effects of Polymethoxyflavone on Photoaging and Its Molecular Mechanism

吉﨑, 舟洋 筑波大学 DOI:10.15068/00160455

2020.07.22

概要

Sun light causes the hyper-pigmentation and the wrinkle formation on the skin. Therefore, they are collectively referred as “photoaging”. Ultraviolet (UV) B is the lowest wavelength in the sunlight arriving at the surface of earth and mainly contributes to the photoaging. Of the photoaging, the hyper-pigmentation results from over– production of melanin in the skin, which is produced by melanocytes of epidermis. On the other hands, the wrinkle is formed by degradation of extracellular matrix (ECM) of the dermis. In these studies, I have examined the effects of polymethoxyflavone (PMF), extracted from orange peels, against the UVB-induced melanogenesis and ECM degradation.

I have firstly focused the UVB-induced prostaglandin (PG)E2 production in the skin. PGE2 is mainly secreted by UVB-irradiated keratinocytes of the epidermis and takes the central role in the skin inflammation, as the erythema and edema formation. Skin inflammation promotes the invasion of inflammatory cells, as lymphocytes and neutrophils, to the dermis. Inflammatory cells secrete ECM degrading enzymes, as matrix metalloproteinase (MMP)s. Moreover, PGE2 also promotes the melanin production and melanin transfer to keratinocytes through the dendrite elongation of melanocytes. Therefore, because I have thought that PGE2 also plays a key role in the UVB-induced melanogenesis and ECM degradation, I have examined the PMF effect against the PGE2 production of HaCaT, human keratinocyte cell line. Firstly, I have found that PMF significantly suppresses the UVB-induced PGE2 production on HaCaTs. PGE2 is synthesized from arachidonic acid by several enzymes. Cyclooxygenase (COX)-2 is the rate-limiting enzyme of this reaction and its expression is induced by UVB. Similar to PGE2, PMF has also inhibited UVB–induced COX-2 mRNA and protein. It has been reported that COX-2 expression is suppressed by peroxisome proliferator–activated receptor (PPAR)-γ activation. By reporter gene assays, in PPAR-α, γ and δ, it has revealed that PMF strongly activates only PPAR-γ. Additionally, PPAR-γ antagonist has recovered the inhibitory effect of PMF against COX-2 expression. Therefore, it has been suggested that PMF suppresses the UVB-induced COX-2 expression and PGE2 production through PPAR-γ activation.

It has been known that UVB also induces ECM degradation, independent on inflammation. In that event, keratinocytes, the major cells in the skin, mainly secretes MMPs. I have focused MMP-1, degrading type 1 collagen which is the most abundant ECM in the dermis. Firstly, I have found that PMF suppresses the UVB-induced MMP-1 expression on HaCaTs. It has been known that MMP-1 expression is activated by NF-κB and activator protein 1(AP-1). Although BAY11-7085, NF-κB inhibitor, has not affected the MMP-1 suppression by PMF, only SP600125, AP-1 inhibitor, has recovered it. It has been reported that AP-1 is activated by phosphorylated c-jun N-terminal kinase (JNK). UVB-induced JNK phosphorylation has been inhibited by PMF treatment. Therefore, it has been suggested that PMF suppresses MMP-1 expression by the inhibition of JNK phosphorylation.

Next, I examined the inhibitory effect of PMF against the melanogenesis. Firstly, I have found that PMF suppresses the melanin production on HM3KO, human-derived melanoma cell line. Melanin is synthesized from tyrosine by several enzymes in the melanosomes of melanocytes. Because tyrosinase is the rate-limiting enzyme of the melanin synthesis, I have examined the effects of PMF against tyrosinase. Although PMF has not affected the enzymatic activity and mRNA expression of tyrosinase, it has decreased tyrosinase protein. Tyrosinase is degraded in proteasomes and lysosomes. Of them, only lysosome inhibitor has recovered tyrosinase degradation by PMF. However, lysosome inhibitor has not recovered the suppression of the melanogenesis by PMF. It has been reported that melanin synthesis is strongly affected by melanosomal pH. I have revealed that PMF acidifies cellular organelles and its majority is the melanosomes. Moreover, ammonium chloride, neutralizing melanosomes, has recovered the suppression of the melanin production and tyrosinase mis-localization by PMF. Therefore, it has been suggested that PMF suppresses the melanin production and tyrosinase localization to the melanosomes by the melanosomal acidification. It has been thought that mis-localized tyrosinase is degraded by the lysosomes.

From above results, I suggest that PMF suppresses the UVB-induced wrinkle formation and hyper-pigmentation in the skin through the suppressions of the PGE2 production, JNK phospholyration and melanogenesis., respectively. Therefore, I think that PMF is inclusively the useful agent against the wrinkle formation and hyper-pigmentation, which are the features of photoaging.

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