Journal of the Society of Cosmetic Scientists of Korea (대한화장품학회지)

Society of Cosmetic Scientists of Korea (대한화장품학회)
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Antioxidant Effects of Cysteine-containing Peptides of Different Lengths in Human HaCaT Keratinocytes Exposed to Hydrogen Peroxide

과산화수소에 노출된 인간 각질형성세포에서 길이가 다른 시스테인 함유 펩타이드의 항산화 효과

  • Jae Won Ha (BK21 PLUS KNU Biomedical Convergence Program for Creative Talent, Department of Molecular Medicine, Cell and Matrix Research Institute, School of Medicine, Kyungpook National University) ;
  • Joon Yong Choi (BK21 PLUS KNU Biomedical Convergence Program for Creative Talent, Department of Molecular Medicine, Cell and Matrix Research Institute, School of Medicine, Kyungpook National University) ;
  • Yong Chool Boo (BK21 PLUS KNU Biomedical Convergence Program for Creative Talent, Department of Molecular Medicine, Cell and Matrix Research Institute, School of Medicine, Kyungpook National University)
  • 하재원 (경북대학교 의과대학, BK21 플러스 KNU 의생명융복합 창의인재양성 사업단, 분자의학교실 세포기질연구소) ;
  • 최준용 (경북대학교 의과대학, BK21 플러스 KNU 의생명융복합 창의인재양성 사업단, 분자의학교실 세포기질연구소) ;
  • 부용출 (경북대학교 의과대학, BK21 플러스 KNU 의생명융복합 창의인재양성 사업단, 분자의학교실 세포기질연구소)
  • Received : 2023.05.30
  • Accepted : 2023.08.24
  • Published : 2023.09.30
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Abstract

Hydrogen peroxide (H2O2) is a type of active oxygen species (ROS) that causes oxidative stress in cells and affects cell growth, proliferation, senescence, and death. The purpose of this study is to find active peptides that attenuate cytotoxicity of H2O2. A positional scanning synthetic tetrapeptide combinatorial library was screened to predict the sequence of potentially active peptides. As a result of comparing the effect of peptide pools on H2O2-induced death of human keratinocytes (HaCaT cells), various active peptide sequences were predicted. Especially, peptides containing cysteine (C) residue were predicted to be active. In follow-up experiments, the cytotoxicity and activity of cysteine-containing peptides of different lengths, such as C-NH2, CC-NH2, CCC-NH2, and CCCC-NH2 were examined. C-NH2 and CC-NH2 showed no significant cytotoxicity up to 1.0 mM, but CCC-NH2, and CCCC-NH2 showed relatively strong cytotoxicity. C-NH2 and CC-NH2 alleviated H2O2-induced cytotoxicity. CC-NH2 was more cytoprotective compared to C-NH2, C, N-acetyl cysteine (NAC), and glutathione (GSH). When intracellular ROS was measured by flow cytometry, H2O2 increased ROS production, and CC-NH2 suppressed ROS production more effectively than C-NH2, and it was as effective as C, NAC, and GSH. This study suggests that CC-NH2 of the cysteine-containing peptides of different lengths has an antioxidant property that safely and effectively alleviates H2O2-induced cytotoxicity and ROS production.

H2O2는 세포에서 산화 스트레스를 유발하여 세포 성장, 증식, 노화 및 사멸에 영향을 미치는 일종의 활성산소종(ROS)이다. 본 연구의 목적은 H2O2의 세포 독성을 완화시키는 활성 펩타이드를 찾는 것이다. 잠재적인 활성 펩타이드의 서열을 예측하기 위해서 위치 스캐닝 합성 테트라펩타이드 조합 라이브러리를 탐색하였다. H2O2로 유도된 인간 각질형성세포(HaCaT 세포)의 사멸에 대한 펩타이드 풀들의 완화 효과를 비교한 결과, 다양한 활성 펩타이드의 시퀀스가 예측되었다. 특히 시스테인(C) 잔기를 함유한 펩타이드가 활성이 있을 것으로 예측되었다. 후속 실험에서 C-NH2, CC-NH2, CCC-NH2, CCCC-NH2 등의 길이가 다른 시스테인 함유 펩타이드들의 세포 독성과 활성을 조사하였다. C-NH2 및 CC-NH2는 1.0 mM 이하에서 유의한 세포 독성이 없었지만 CCC-NH2 및 CCCC-NH2는 상대적으로 강한 세포 독성을 보였다. C-NH2와 CC-NH2는 H2O2로 유도된 세포독성을 완화시켰다. CC-NH2는 C-NH2, C, N-아세틸 시스테인(NAC) 및 글루타티온(GSH)보다 세포 보호 효과가 높았다. 유세포 분석법으로 세포 내 ROS를 측정하였을 때, H2O2는 ROS의 생성을 증가시켰다.H2O2 노출조건에서 CC-NH2는 C-NH2보다는 더 효과적으로 ROS 생성을 억제하였고, C, NAC, GSH 만큼 효과적이었다. 본 연구의 결과는 다양한 시스테인 함유 펩타이드 중 특히 CC-NH2가 H2O2로 유도된 세포 독성과 ROS 생성을 안전하고 효과적으로 완화시키는 항산화 특성이 있음을 시사한다.

Keywords

Acknowledgement

본 연구는 보건복지부의 재원으로 한국보건산업진흥원의 보건의료기술연구개발사업 지원에 의하여 이루어진 것임(과제고유번호 HP20C0004).

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