UVA가 유도하는 산화적 스트레스에 Ellagic Acid가 미치는 인간 모유두 세포 보호효과
Protective Effects of Ellagic Acid against UVA-induced Oxidative Stress in Human Dermal Papilla
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목적: 태양광을 구성하는 성분 중 하나인 UVA는 피부에서 활성산소(reactive oxygen species, ROS를) 생산하고 피부 깊숙히 침투할 수 있는 능력을 가지고 있다. 이러한 특성은 진피층에 광분해생성물이나 세포노화, 세포 사멸, 염증, 유전자 발현 변화와 같은산화적 스트레스를 유발한다. 따라서, UVA는 피부를 자극하는 주요 외부인자로 여겨지고 있다. 본 연구의 목적은 UVA에 의한자극에 ellagic acid의 항산화 및 항염 효과를 인간 모유두 세포를 이용하여 확인하였다. 방법: 세포 생존력은 WST-1 방법을 사용하여 수행하였으며, DCFH-DA 을 이용하여 세포 내 활성산소 소거능을 연구하였다. 본 연구에서 시험한 모든 mRNA 발현은 qRT-PCR을 통해 분석하였으며 세포노화는 SA-β-gal 세트를 사용하여 조사하였다. 세포이동능 분석을 통해 ellagic acid가 HDPC의 이동능에 미치는 영향을 알아보았다. 결과: Ellagic acid 25 μM까지 세포독성은 확인되지 않았으며, UVA로 인한 세포독성에 ellagic acdi가 보호하는 효과를 확인하였다. 더불어, 항산화 효소인 CAT의 유전자 발현변화를 유도하는 효과와 세포 내 활성산소를 소거하는 효과를 실험을 통해 입증하였다. 이뿐만 아니라, UVA 조사로 인해 발현이 증가한 염증유도인자, TNF-α, IL-6의 mRNA 발현 수준을 조절하는 것으로 나타났으며UVA로 인해 손상된 세포의 세포 이동능을 보호하는 것에도 유의적인 효과가 있음을 확인할 수 있었다 . 결론: 본 연구결과는 ellagic acid가 UVA로 인해 유도되는 산화적 스트레스를 보호하는 효과를 인간 모유두 세포를 이용하여 입증하였다.
Abstract
Purpose: One of solar radiation components, ultraviolet A (UVA), has deeply penetrating capacity and generates reactive oxygen species (ROS) in the skin. These characteristics arouse oxidative stress in dermis, such as photoproducts, senescence, cell death, inflammation, and alterative gene expression. Thus, UVA is considered a major factor of external stimuli that irritates the skin. This study aimed at investigating the antioxidant and anti-inflammatory effects of ellagic acid on UVA-irradiated human dermal papilla cells (HDPCs). Methods: The cell viability of HDPCs were analyzed using the water-soluble tetrazolium salt (WST-1) assay. 2,7-dichlorofluorescin diacetate (DCFH-DA) was used in evaluating intracellular ROS scavenging activity. All the mRNA expression levels were carried out via quantitative real-time polymerase chain reaction (qRT-PCR). Cellular senescence was evaluated using senescence-associated β-galactosidase (SA-β-gal) staining kit. The wound healing assay in HDPCs were performed to assess effects of ellagic acid on the cell migration activity. Results: No cell cytotoxicity of ellagic acid was observed at concentration up to 25 μM. In irradiated HDPCs, ellagic acid showed protective effects on UVA-induced cytotoxicity. Further, ellagic acid represented ROS scavenging activity and regulating antioxidant gene expression, catalase transcriptional levels under UVA-irradiated condition. It also modulated tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) mRNA expresison levels and cellular senescence on UVA-stimulated HDPCs. Cell migration assay showed the protective effects of ellagic acid on movement of HDPCs damaged by UVA. Conclusion: These results demonstrate that ellagic acid protects UVA-induced oxidative stress in HDPCs.
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