메틸말리올라이드의 NRF2/ARE 유도를 통한 피부 세포 보호 효과

Skin Protective Effect of Methylated Marliolide through Induction of NRF2/ARE

Abstract

본 연구진은 메틸말리올라이드가 인간 피부 세포주인 HaCaT 세포에서 NRF2를 유도하고 이를 통하여 항산화 효과를 나타내는지 알아보았다. MTT assay를 통하여 메틸말리올라이드는 $10{\mu}M$ 농도에서 24 h 동안 HaCaT 세포에 처리하여도 HaCaT 세포 독성을 나타내지 않는 것을 확인하였다. 본 연구진이 구축한 HaCaT-ARE-GFP-luciferase 세포에 메틸말리올라이드를 처리하고 루시퍼라아제 활성을 측정한 결과 메틸말리올라이드는 양성 대조군인 레스베라트롤보다 ARE 결합에 따른 루시퍼라아제 활성을 더욱 강하게 증가시키는 것을 확인하였다. 또한 HaCaT 세포에 메틸말리올라이드 처리 시 NRF2에 의하여 유도되는 항산화 단백질인 HO-1과 NQO1 mRNA 및 단백질 발현이 통계적으로 유의하게 증가하였다. 마지막으로 메틸말리올라이드는 HaCaT 세포에서 TPA에 의해서 유도된 DNA 및 지질의 산화를 강력하게 억제하였다. 결론적으로 본 연구는 메틸말리올라이드가 NRF2 유도를 통하여 피부 산화적 스트레스를 억제하며 이는 메틸말리올라이드가 신규 항산화 화장품의 소재로 적합하다는 것을 시사한다.

In the present study, we have investigated whether methylated marliolide could induce NRF2 thereby exerting anti-oxidant effects. MTT assay showed that methylated marliolide did not exhibit cytotoxicity on HaCaT cells. Methylated marliolide induced a higher ARE-dependent luciferase activation in HaCaT ARE-GFP-luciferase cells, compared with resveratrol. In addition, exposure of methylated marliolide to HaCaT cells significantly induced NRF2 and transcriptionally activated HO-1 and NQO1, both of which are target genes of NRF2. Finally, methylated marliolide protected HaCaT cells against TPA-induced oxidative damages on nucleotides and lipids. Together, results shows that methylated marliolide could suppress oxidative damages through induction of NRF2 which implies that methylated marliolide might serve as a good candidate for novel cosmetic ingredient with anti-oxidant effects.

Figure 1. A single step synthesis of methylated marliolide from marliolide whose synthetic scheme was previously reported[8].

Figure 2. MTT assay shows that methylated marliolide does not affect the viability of HaCaT cells.

Figure 3. Luciferase assay shows that methylated marliolide significantly activates ARE-luciferase activity in HaCaT-ARE-GFP-luciferase cells. Resveratrol was included as a positive control.

Figure 4. Western blot shows that methylated marliolide induces HO-1, NQO1, and NRF2 in HaCaT cells. Resveratrol was included as a positive control.

Figure 5. Methylated marliolide transcriptionally induces HO-1 and NQO1 in HaCaT cells.

Figure 6. Immunofluorescence study shows that methylated marliolide suppress TPA-induced generation of 4-hydroxynonenal (4-HNE, Left Panel) and 8-oxodeoxyguanosine (8-OHdG, Right Panel) in HaCaT cells.