The present study aimed to investigate the anti-inflammatory and anti-melanogenesis activity of Helichrysum bracteatum flower extracts in vitro. H. bracteatum flowers were extracted with water, ethanol, and 1,3-butylene glycol, and the anti-oxidative activities of the extracts were measured using a 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The inhibition of the expression of inflammation-related genes, including tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and cyclooxygenase-2 (COX-2), were evaluated in vitro using reverse transcription (RT)-PCR. To investigate the inhibitory effects of H. bracteatum flower extracts on inflammation, the production of TNF-α and nitric oxide (NO) in human epidermal keratinocytes (HEKa cells) were measured using enzyme-linked immunosorbent assays. The inhibition of the expression of melanogenesis-related genes, including tyrosinase (TYR), tyrosinase-related protein (TRP)-1, TRP-2, and microphthalmia transcription factor (MITF) were measured in vitro using RT-PCR and the production of cAMP and PKA in melanoma cells (SK-MEL2). To evaluate the inhibitory effects of H. bracteatum flower extracts on melanogenesis, the production of melanin in SK-MEL2 cells and mushroom tyrosinase inhibition effects were measured. Also, the gene expression of POMC in epidermal keratinocyte (HEKa cells) was measured using RT-PCR. The DPPH assay results showed that H. bracteatum flower extracts have good anti-oxidative effects. H. bracteatum flower extracts inhibited the expression of inflammation-related genes TNF-α, IL-6, and COX-2. Moreover, the production of TNF-α and NO were inhibited by H. bracteatum flower extracts. The expression of melanogenesis-related genes TYR, TRP-1, and TRP-2 were inhibited by H. bracteatum flower extracts. Additionally, the productions of cAMP and PKA were inhibited by H. bracteatum flower extracts in melanoma cells (SK-MEL2). Melanin synthesis and mushroom tyrosinase activity inhibited with H. bracteatum flower extracts. POMC gene expression in keratinocyte (HEKa cells) was also inhibited by H. bracteatum flower extracts. These results suggest that H. bracteatum flower extracts have the potential for use as a cosmetic raw material with anti-inflammation efficacy which inhibition of TNF-ɑ dominantly and also suppress NO production and inflammation-related genes such as TNF-ɑ, IL-6, and COX-2. Moreover, H. bracteatum flower extracts could be used as a whitening agent with a good tyrosinase inhibition activity and/or inhibition of signals of melanogenesis.
칠채국추출물 (Helichrysum bracteatum flower extracts)의 화장품 원료로서 가능성을 확인하기 위하여 항염 효과와 미백 효과를 피부세포를 이용하여 평가하였다. 칠채국을 물, 알코올, 1,3-butyleneglyol을 용매로 추출하고 각각의 추출물의 항산화 효과를 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay로 평가한 결과 95% 알코올을 용매로 추출한 경우 항산화 효과가 가장 우수하였다. Reverse transcription (RT)-PCR 분석 결과, 칠채국 추출물은 염증과 관련된 유전자인 tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) 및 cyclooxygenase-2 (COX-2)의 발현을 저해하였으며, enzyme-linked immunosorbent assays를 통해 nitric oxide (NO)와 TNF-α의 생성도 저해됨을 확인하였다. 뿐만 아니라, 칠채국 추출물은 항염 효과와 더불어 미백소재로의 활용 가능성도 보여주었다. RT-PCR 분석 결과, 칠채국 추출물 처리 시, Microphthalmia associated transcription factor (MITF), tyrosinase (TYR), tyrosinase related protein (TRP)-1, 및 TRP-2의 발현이 농도 의존적으로 감소하였으며, cAMP 및 PKA의 생성 또한 저해됨을 확인하였다. 이는 칠채국 처리 흑색종(SK-MEL2)에서의 멜라닌 생성량 분석을 통해 다시 한번 확인하였으며, mushroom tyrosinase의 활성 또한 칠채국 추출물에 의해 저해되는 결과를 확인하였다. 칠채국 추출물은 멜라닌세포에서 뿐만 아니라, 각질형성세포(HEKa)에서 POMC의 발현을 저해함으로 멜라닌 세포에서의 색소생성 저해 가능성을 보여주었다. 이상의 결과를 종합할 때, 칠채국 추출물은 TNF-α의 조절로부터 시작되는 항염 효과 및 tyrosinase 활성을 저해하며, 보조적으로 멜라닌 생성 기작의 지표인자 발현 저해를 통한 미백 효과를 보여주는 기능성 화장품의 소재로 활용 가능할 것으로 사료된다.