인간 진피섬유아세포에서 Chlorella sp. HS1 유래 오일의 콜라겐 발현 증가 효력에 관한 연구
Potential Anti-wrinkle Activity of Chlorella sp. HS1-derived Oil Components on Human Dermal Fibroblasts
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목적: 본 연구는 Chlorella sp. HS1 유래 오일의 진피섬유아세포 내에서 콜라겐 발현에 관한 효과를 분석하고자 하였다. 방법: Chlorella sp. HS1 바이오매스로부터 오일을 추출한 후, 고성능액체크로마토그래피 분석을 통해 추출물 내의 성분 종류와 함량을 분석하였다. 인간 진피섬유아세포를 대상으로, 오일 추출물 처리에 따른 세포 생존율과 정량적 RT-PCR분석을 통한 콜라겐과 콜라겐분해효소 발현량을 분석하였다. 뿐만 아니라, 오일 추출물에 의한 콜라겐 발현 효과가 지표 성분인 베타카로틴에 의한 것인지에 대하여도 분석하였다. 또한, 자외선이 조사된 진피섬유아세포의 보호효능에 대한 베타카로틴의 효과에 대해서도 분석하였다. 결과: 오일 추출물 50 μg/mL 이하의 농도에서 각질형성세포, 멜라닌형성세포, 진피섬유아세포의 세포 생존율에는 영향이 낮음을 보였다. 고성능액체크로마토그래피 분석을 통해, 오일 추출물 내 베타카로틴이 가장 주요한 성분으로 나타났으며, 베타카로틴을 진피섬유아세포 처리시 낮은 세포독성을 나타냈다. 추가적인 실험을 통해 오일 추출물 처리시 콜라겐 타입 1과 타입 3의 발현이 현저하게 증가하였으며, MMP1과 MMP3의 발현은 현저하게 감소되었다. 이러한 효과는 베타카로틴을 처리한 세포에서도 동일하게 나타났다. 또한, 베타카로틴은 자외선에 의해 유도되는 콜라겐 타입 1과 타입 3의 발현 감소를 현저하게 회복시키는 효과가 나타났다. 결론: 본 연구결과를 통해 베타카로틴이 함유된 Chlorella sp. HS1 유래 오일의 주름개선 성분의 가능성을 제시하였다.
Abstract
Purpose: In this study, we investigated the effects of Chlorella sp. HS1-derived oils on collagen expressions in dermal fibroblasts. Methods: We prepared the oil extracts from Chlorella sp. HS1 biomass, and high-performance liquid chromatography analysis was used in the identification and quantitation of components in the oil extracts. Using human dermal fibroblasts, cell viability and quantitative RT-PCR (qRT-PCR)-mediated expression analysis of collagen and matrix metalloprotease (MMP) genes after oil treatments. We further investigated whether the effect of the oil extracts on collagen expression was related to the major component, β-carotene, of the extracts. In addition, we analyzed the effect of photoprotective effect of β-carotene on ultraviolet (UV)-irradiated dermal fibroblasts. Results: We found that under 50 μg/mL of the oil extracts had little cytotoxicity in skin cells including epidermal keratinocytes, melanocytes and dermal fibroblasts. The HPLC analysis revealed that β-carotene was identified as the major non-polar molecules. Next, we found that the oil-derived β-carotene was little cytotoxicity in dermal fibroblasts. Further experiments revealed that treatment with the oil extracts significantly upregulated collagen type 1 and 3 mRNAs and downregulated MMP1 and MMP3 mRNAs in dermal fibroblasts. Also, we found that these effects were showed in β-carotene-treated cells. In addition, β-carotene significantly attenuated UVA-induced downregulation of collagen type 1 and 3 expressions. Conclusion: Therefore, our results suggested that the oil extracts had a potential anti-wrinkle agent via its major component, β-carotene.
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