[논문]방사무늬 김 열수추출물의 RAW 264.7 세포에서의 면역 증진 효과

장고은; 박보람; 이슬아; 김춘성

doi:10.15433/ksmb.2023.15.2.033

[국내논문] 방사무늬 김 열수추출물의 RAW 264.7 세포에서의 면역 증진 효과
Immune Enhancing Effects of Pyropia yezoensis Hydrothermal Extract in RAW 264.7 Cells 원문보기

한국해양바이오학회지 = Journal of marine bioscience and biotechnology, v.15 no.2, 2023년, pp.33 - 40

장고은 (해양헬스케어 유효성실증센터) , 박보람 (경운대학교 보건복지대학 치위생학과) , 이슬아 (해양헬스케어 유효성실증센터) , 김춘성 (해양헬스케어 유효성실증센터)

Abstract ▼ AI-Helper

This study aimed to investigate the immunomodulatory function of Pyropia yezoensis hydrothermal (water) extract (PYWE) in comparison to the group treated only with lipopolysaccharides (LPS) in RAW264.7 cells. LPS is known to be an inflammatory mediator that activates macrophages, leading to the secretion of nitric oxide (NO), inducible nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) as defense responses. Through enzyme-linked immunoassay and western blot analyses, it was observed that PYWE increased the expression levels of NO, iNOS, TNF-α, and IL-6 in RAW264.7 cells in a dose-dependent manner, although to a lesser extent compared with the group treated with LPS alone. In addition, the study examined the mitogen-activated protein kinases (MAPKs) pathway, which regulates various cellular activities, including gene expression, mitosis, cell differentiation, transformation, survival, and death. The western blot analysis confirmed that PYWE also regulated the MAPKs pathway. Furthermore, the expression levels of immunomodulatory-related factors increased in the group treated with PYWE compared with the control group. Even though the effects of PYWE were usually less strong than those of LPS, the effects of PYWE increased with increasing doses compared to the control group. This suggests that PYWE could be used to control the immune system.

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표/그림 (4)

그림 Fig. 1. Cell viability of PYWE on RAW264.7 cells. RAW264.7 cells were treated with PYWE (0.125, 0.25, 0.5, 1 mg/mL) for 24 h, and cell viability was measured by MTT assay. RAW264.7 cells were incubated without PYWE as control group, which were considered 100% survival. Data are represented as mean ± SD of three independent experiments. N = 3 per group.
그림 Fig. 2. Effects of PYWE on the expression levels of Nitric oxide, iNOS, TNF-α, and IL-6 in RAW264.7 cells. RAW264. 7 cells were pre-treated with PYWE (0.125, 0.25, 0.5, 1 mg/m L) for 24 h, and LPS (0.5 μg/mL) was respectively induced for inflammatory response for 24 h. (a) NO production was detected by using the supernatant of cultured medium and Griess reagents. (b) Protein expression levels of the iNOS, TNF-α, and IL-6 was examined using Western blot analysis. α-Tubulin represented as a control. N = 3 per group. Data are served as mean ± SD of three independent experiments. ** p < 0.05, and *** p < 0.005 compared with the Control group.
그림 Fig. 3. Effects of PYWE on the cytokine expression levels of TNF-α, and IL-6 in RAW264.7 cells. RAW264.7 cells wer e pre-treated with PYWE (0.125, 0.25, 0.5, 1 mg/mL) for 24 h, and LPS (0.5 μg/mL) was respectively induced for infla mmatory response for 24 h. (a) TNF-α concentration was dete cted by ELISA kits using the supernatant of cultured medium. (b) IL-6 concentration was examined by ELISA kits using the supernatant of cultured medium. N = 3 per group. Data are served as mean ± SD of three independent experiments. ** p < 0.05, and *** p < 0.005 compared with the Control group.
그림 Fig. 4. Effects of PYWE on the protein expression levels of MAPKs in RAW264.7 cells. RAW264.7 cells were pre-tre ated with PYWE (0.125, 0.25, 0.5 mg/mL) for 24 h, and LPS (0.5 μg/mL) was respectively induced for inflammatory response for 24 h. Protein expression levels of the ERK, p-ER K, JNK, p-JNK, P38, and p-P38 was measured by Western blot analysis. (b-d) Quantitative data of (a) were analyzed using the ImageJ bundled with Java 1.8.0_172 software. N = 3 per group.

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