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RAW 264.7 대식세포에서 lipopolysaccharide 자극에 의한 염증성 및 산화적 스트레스에 미치는 5-aminolevulinic acid phosphate의 영향
지선영(Seon Yeong Ji),김민영(Min Yeong Kim),황보현(Hyun Hwangbo),이혜숙(Hyesook Lee),홍수현(Su Hyun Hong),차희재(Hee-Jae Cha),김희수(Heui-Soo Kim),김석만(Suhkmann Kim),최영현(Yung Hyun Choi) 한국생명과학회 2021 생명과학회지 Vol.31 No.9
5-ALA-p는 천연 아미노산인 5-ALA를 암모니아수로 용출하고 인산과 아세톤을 첨가하여 광역학 요법에 적합한 특성을 갖도록 개발된 물질이다. 그러나 항산화 및 항염증에 대한 잠재적인 기전을 포함한 약리학적 효능은 아직 명확하지 않다. 본 연구에서는 LPS로 자극된 RAW 264.7 세포에서 산화적 및 염증성 반응에 대한 5-ALA-p의 효과를 평가하였다. 본 연구의 결과에 의하면, 5-ALA-p는 LPS에 의한 RAW 264.7 세포의 과도한 식균 활성을 유의하게 억제하였고 산화적 스트레스를 약화시켰다. 5-ALA-p는 또한 LPS에 의해 감소된 미토콘드리아 생물 발생을 개선하였으며, 이는 5-ALA-p가 LPS로 인한 미토콘드리아 손상을 복원시켰음을 시사한다. 아울러 5-ALA-p는 NO와 TNF-α, IL-1β 및 IL-6과 같은 염증성 사이토카인의 생성을 현저히 억제하였으며, 이는 iNOS 및 각 사이토카인의 발현 감소와 연관성이 있었다. 나아가 5-ALA-p는 NF-κB의 핵 전이를 감소시키고 MAPKs의 인산화를 억제하여 5-ALA-p의 항염증 효과가 이들 신호전달 경로의 활성 억제와 매개되었음을 보여주었다. 이러한 결과들은 5-ALA-p가 산화적 및 염증성 스트레스를 줄이는 잠재적인 후보 약물로 적용될 수 있음을 의미한다. 5-Aminolevulinic acid phosphate (5-ALA-p) is a substance obtained by eluting 5-ALA (a natural delta amino acid) with aqueous ammonia, adding phosphoric acid to the eluate, and then adding acetone to confer properties suitable for use in photodynamic therapy applications. However, its pharmacological efficacy, including potential mechanisms of antioxidant and anti-inflammatory reactions, remains unclear. This study aimed to investigate the effects of 5-ALA-p on oxidative and inflammatory stresses in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Our data showed that 5-ALA-p significantly inhibited excessive phagocytic activity via LPS and attenuated oxidative stress in LPS-treated RAW 264.7 cells. Furthermore, 5-ALA-p improved mitochondrial biogenesis reduced by LPS, suggesting that 5-ALA-p restores mitochondrial damage caused by LPS. Additionally, 5-ALA-p significantly suppressed the release of nitric oxide (NO) and pro-inflammatory cytokines, such as tumor necrosis factor α (TNF-α), interleukin (IL)-1β, and IL-6, which are associated with the inhibition of inducible NO synthase and respective cytokine expression. Furthermore, 5-ALA-p reduced the nuclear translocation of nuclear factor-kappa B (NF-κB) and inhibited phosphorylation of mitogen-activated protein kinases (MAPKs), indicating that the anti-inflammatory effect of 5-ALA-p is mediated through the suppression of NF-κB and MAPK signaling pathways. Based on these results, 5-ALA-p may serve as a potential candidate to reduce inflammation and oxidative stress.
잉어의 생체 내 대사체 변화에 미치는 플록세틴 영향규명 연구
정인영 ( In-young Chung ),박유미 ( Yu-mi Park ),김상민 ( Sang-min Kim ),김일규 ( Il-gyu Kim ),도영선 ( Young-sun Do ),김석만 ( Suhkmann Kim ),한상범 ( Sang Beom Han ),조현덕 ( Hyun-deok Cho ),석광설 ( Kwang-seol Seok ) 한국환경분석학회 2017 환경분석과 독성보건 Vol.20 No.1
The purpose of this study was to clarify the effect of fluoxetine, one of the most frequently prescribed antidepressant drugs, on Cyprinus carpio by comparing the variations in its metabolite concentrations after exposure to the drug, using NMR and LC/Orbitrap MS. Three experimental reactors were employed, and the total experimental period was 14 days. Cyprinus carpio were acclimatized for 3 months in the laboratory and then subjected to the study. One reactor was used as a 14-day control, and the others were exposed to fluoxetine at 10 ng/L and 10 μg/L for 7 days and depurate for 7 days. The variations in metabolite concentration after exposure were analyzed by NMR and LC/Orbitrap MS. Before exposure, the concentration trends of fluoxetine and its main metabolite, norfluoxetine were almost the same in blood and liver samples of Cyprinus carpio. However, after the depuration period, the concentrations of norfluoxetine in the liver and blood samples were higher than those of fluoxetine. The concentration of the remaining norfluoxetine in the liver sample was higher than that in the blood sample. This means that the liver mainly metabolized pharmaceuticals. The main change in the concentrations of metabolites of Cyprinus carpio was shown in amino acids such as leucine, isoleucine, glutamate, tyrosine, and valine; organic acids such as acetate and formate; creatine; and glucose. Although after the depuration period, the control, and 10 ng/L and 10 μg/L exposure groups had overlapping results, the exposure and control groups could be classified clearly after 14 days. These results are expected to contribute to identifying the effects of environmental pollutants by tracing metabolite changes and building a chemical metabolite database.