Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Effects of Paprika Extract and Its Components on Cell Death and Expression of p53 and GADD45 Genes in Ultraviolet B- Exposed HaCaT Cells

UVB를 조사한 HaCaT 세포의 세포사멸과 p53 및 GADD45 유전자 발현에 대한 파프리카 추출물 및 성분들의 효과

  • Ha, Se-Eun (Division of Biological Science and Research Institute for Basic Science, Wonkwang University) ;
  • Kim, Hyung-Do (Division of Biological Science and Research Institute for Basic Science, Wonkwang University) ;
  • Kang, Jea-Ran (Division of Biological Science and Research Institute for Basic Science, Wonkwang University) ;
  • Park, Jong-Kun (Division of Biological Science and Research Institute for Basic Science, Wonkwang University)
  • Received : 2011.03.17
  • Accepted : 2011.05.19
  • Published : 2011.05.30
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Abstract

In the present study, the effects of paprika extract and its components including vitamin C, lycopene and beta-carotene on cell death in ultraviolet B (UVB)-exposed HaCaT cells were investigated. The cell viability upon treatment for 24 hr with either paprika extract or vitamin C alone was similar to or greater than that of the untreated control. However, the viability of the cells treated with lycopene or beta-carotene decreased to about 20% of that in the untreated control. When UVB-exposed cells were post-incubated for 24 hr in medium containing paprika extract or vitamin C, cell viability increased in a concentration dependent manner as compared to those post-incubated in a normal growth medium. In contrast, post-incubation of UVB-exposed cells with lycopene or beta-carotene decreased cell viability in a concentration dependent manner as compared to those post-incubated in a normal growth medium. The nuclear fragmentation analysis showed that paprika extract or vitamin C decreases UVB-induced apoptosis. The apoptotic nuclear fragmentation resulting from UVB exposure was also protected by the paprika extract or vitamin C post-incubation. However, the UVB-induced apoptotic nuclear fragmentation of the cells treated with lycopene or beta-carotene increased in a concentration dependent manner. Western blot analysis showed that either paprika extract or vitamin C treatment alone did not significantly change the level of p53 and GADD45 protein. Interestingly, post-incubation of UVB-exposed cells with paprika extract or vitamin C decreased the p53 and GADD45 protein level as compared to those post-incubated in a normal growth medium. In contrast, incubation of UVB-exposed or non-irradiated cells with lycopene or beta-carotene increased the p53 and GADD45 protein levels in a concentration dependent manner as compared to those incubated in a normal growth medium. All these results suggest that paprika extract and vitamin C help the survival of the UVB-exposed cells, while lycopene and beta-carotene potentiate the apoptotic death of UVB-exposed cells, in accordance with the respective changes in p53 and GADD45 protein levels.

본 연구는 파프리카 추출물과 그 성분인 비타민 C, 라이코펜과 베타-카로틴이 UVB (ultraviolet-B)에 의한 유전독성의 감소에 효과를 보이는 지를 HaCaT 세포를 이용하여 분석하였다. 자외선을 조사하지 않은 정상 세포의 세포활성은 파프리카 추출물을 경우 처리하지 않은 대조군과 차이를 나타내지 않았지만 비타민 C의 경우 농도 의존적으로 증가시키는 것을 관찰하였다. 그러나 라이코펜과 베타-카로틴의 경우 농도 의존적으로 점차 세포활성이 감소하는 것으로 관찰되었다. UVB로 상해받은 세포를 추출물이나 그 성분으로 후 배양할 경우 정상 배양액으로 배양한 대조군에 비해 파프리카 추출물과 비타민C은 농도 의존적으로 세포 활성을 증가시켰으나 라이코펜과 베타-카로틴의 경우 농도 의존적으로 감소키는 것을 관찰할 수 있었다. 자외선을 조사하지 않은 정상 세포의 핵 분절율은 파프리카 추출물과 비타민 C의 경우 대조군과 유의적인 차이를 나타내지 않았지만, 라이코펜, 베타-카로틴의 경우 농도 의존적으로 핵 분절율이 증가하는 것을 관찰하였다. UVB를 조사한 후 파프리카 추출물이나 비타민 C를 처리한 경우 정상 배양액으로 배양한 대조군에 비해 핵 분절율을 감소시켰지만 라이코펜과 베타-카로틴의 경우 농도 의존적으로 증가시켰다. 세포 상해에 반응하는 유전자인 p53과 GADD45의 단백질 수준을 Western blot으로 분석한 결과, 파프리카 추출물만 처리했을 경우 p53과 GADD45 단백질 수준은 처리하지 않은 대조군과 비교해서 유의미한 차이를 나타내지 않았으나 UVB를 조사한 후 파프리카 추출물을 처리한 경우 농도 의존적으로 p53 단백질 발현량이 감소하였다. 비타민 C를 단독 처리할 경우 대조군에 비해 p53과 GADD45 단백질 발현량은 감소하였으며, UVB를 조사한 후 비타민 C를 처리한 경우에도 농도 의존적으로 p53과 GADD45 단백질 발현량이 감소하는 것을 확인할 수 있었다. 그러나 라이코펜과 베타-카로틴만 단독 처리할 경우 p53과 GADD45 단백질 발현량이 처리하지 않은 대조군에 비해 농도 의존적으로 증가하였다. UVB를 조사한 후 베타-카로틴을 처리한 경우에도 상대적인 고농도에서 p53과 GADD45 단백질 발현량이 증가하는 것을 확인할 수 있었다. 위의 결과를 토대로 파프리카 추출물과 비타민 C는 UVB에 의해 손상된 세포의 세포 독성을 회복하는 효능이 있다고 생각된다. 라이코펜과 베타-카로틴의 경우에는 UVB에 의해 발현된 p53 단백질의 수준이 농도 의존적으로 더욱 증가하는 것으로 보아 UVB에 의한 세포고사가 라이코펜과 베타-카로틴에 의해 강화되는 것으로 생각된다.

Keywords

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