[논문]유산균에 의해 변화된 황기-포스트바이오틱스 다당류가 대식세포에 미치는 영향

김연숙; 신현영; 정원비; 하은지; 구자평; 신지영; 유광원

doi:10.9799/ksfan.2024.37.1.017

유산균에 의해 변화된 황기-포스트바이오틱스 다당류가 대식세포에 미치는 영향
Effect of Astragalus membranaceus-postbiotics Polysaccharide Changed by Lactic Acid Bacteria on Macrophage 원문보기

한국식품영양학회지 = The Korean journal of food and nutrition, v.37 no.1, 2024년, pp.17 - 29

김연숙 (한국교통대학교 식품영양학전공) , 신현영 (고려대학교 대학원 의생명융합과학과 러닝헬스시스템 융합전공) , 정원비 (한국교통대학교 식품영양학전공) , 하은지 (한국교통대학교 식품영양학전공) , 구자평 (한국교통대학교 식품영양학전공) , 신지영 ((주)코시스바이오) , 유광원 (한국교통대학교 식품영양학전공)

Abstract ▼ AI-Helper

To increase industrial applicability of Astragalus membranaceus (AM) as immunostimulating materials, hot-water extract (AME) was prepared from AM and fermented with Kimchi-lactic acid bacteria (Lactobacillus sakei & Leuconostoc mesenteroides) to prepare fermented AM-postbiotics (FAME). Although FAME prepared from AM-postbiotics did not show a significant enhancement in macrophage stimulating activity compared to non-fermented AME, crude polysaccharide (FAME-CP) fractionated by EtOH precipitation from FAME showed significantly higher macrophage stimulating activity than AME-CP. Compared to AME-CP, FAME-CP showed dramatic changes in component sugar and molecular weight distribution. FAME-CP was a polysaccharide with a major molecular weight distribution of 113.4 kDa containing Man (44.2%), Glc (19.3%), Gal (10.2%), GalA (10.2%), and Ara (7.4%) as sugar components. FAME-CP with enhanced macrophage stimulatory activity not only increased expression levels of mRNA genes encoding macrophage-activated factors (iNOS, TNF-α, MCP-1, IL-6, and COX-2), but also led the nuclear translocation of activated p65 and c-Jun. In conclusion, crude polysaccharide from AM-postbiotics fermented with lactic acid bacteria could increase industrial applicability as a functional material with enhanced immunostimulating activity than AME-CP.

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표/그림 (9)

그림 Fig. 1. Fractionation scheme of crude polysaccharide isolated from AME and FAME.
표 Table 1. HPLC analytical condition for component sugar and molecular weight distribution of crude polysaccharide isolated from AME and FAME
표 Table 2. The primer sequence of the mouse for qRT-PCR
그림 Fig. 2. Macrophage stimulating activity of AME and FAME. RAW 264.7 cells were treated with AME and FAME for 24 hr, and evaluated cytotoxicity effect (A), and production of nitric oxide (B), TNF-α (C), MCP-1 (D), and IL-6 (E). Results are expressed as mean±S.D. of three independent tests in triplicate. Different asterisks on the column indicate significant differences between negative control (NC) and each group by Student's t-test. The crosshatch patterns expressed significant differences between samples within the same concentrations. *,^#p<0.05, **,^##p<0.01, ***,^###p<0.001. NC; negative control, LPS; lipopolysaccharide (1 μg/mL); positive control, AME; hot-water extracts from AM, FAME; AM-postbiotics.
그림 Fig. 3. Macrophage stimulating activity of crude polysaccharide isolated from AME and FAME. RAW 264.7 cells were treated with crude polysaccharide isolated from AME and FAME for 24 hr, and evaluated cytotoxicity effect (A), and production of nitric oxide (B), TNF-α (C), MCP-1 (D), and IL-6 (E). Results are expressed as mean±S.D. of three independent tests in triplicate. Different asterisks on the column indicate significant differences between negative control (NC) and each group by Student's t-test. The crosshatch patterns expressed significant differences between samples within the same concentrations. *,^#p<0.05, **,^##p<0.01, ***,^###p<0.001. NC; negative control, LPS; lipopolysaccharide (1 μg/mL); positive control, AME; hot-water extracts from AM, FAME; AM-postbiotics, CP; Crude polysaccharide.
그림 Fig. 4. Molecular weight distribution chromatogram of crude polysaccharide isolated from AME and FAME.
표 Table 3. Chemical property and component sugar of crude polysaccharide isolated from AME and FAME
그림 Fig. 5. Effect of FAME-CP on macrophage stimulating factors-mRNA gene expression in RAW 264.7 cells. RAW 264.7 cells were treated with FAME-CP for 24 hr and evaluated mRNA gene expression of iNOS (A), TNF-α (B), MCP-1 (C), IL-6 (D), and COX-2 (E). Results are expressed as mean±S.D. of three independent tests in triplicate. Different asterisks on the column indicate significant differences between negative control (NC) and each group by Student's t-test. *p<0.05, **p<0.01, ***p<0.001. NC; negative control, LPS; lipopolysaccharide (1 μg/mL); positive control, FAME; AM-postbiotics, CP; crude polysaccharide.
그림 Fig. 6. Effect of FAME-CP on nuclear translocation of activated p65 and cJun in RAW 264.7 cells. RAW 264.7 cells were seeded into 0.1% gelatin-coated coverslip and then treated with FAME-CP for 3 hr. RAW 264.7 cells were fixed and immunostained with FITC conjugated anti-p65 and anti-cJun antibody and the nucleus was counterstained with DAPI. NC; negative control, LPS; lipopolysaccharide (1 μg/mL); positive control, FAME; AM-postbiotics, CP; crude polysaccharide.

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