[논문]도라지 잎 에탄올 추출물의 주요 성분 분석 및 마우스 대식세포와 인체 폐암세포에서 항염효과

이정민; 배병준; 최지림; 정영신

doi:10.9799/ksfan.2024.37.2.110

도라지 잎 에탄올 추출물의 주요 성분 분석 및 마우스 대식세포와 인체 폐암세포에서 항염효과
Analysis of Major Constituents of an Ethanol Extract of Platycodon Grandiflorum Leaves and Protective Effects on Inflammation in Murine Macrophage and Human Lung Carcinoma Cells

한국식품영양학회지 = The Korean journal of food and nutrition, v.37 no.2, 2024년, pp.110 - 122

이정민 (호서대학교 대학원 생명공학과) , 배병준 (호서대학교 대학원 생명공학과) , 최지림 ((주)황초원) , 정영신 (호서대학교 생명공학과)

Abstract ▼ AI-Helper

This study investigated major constituents and anti-inflammatory effects of an ethanol extract of Platycodon grandiflorum leaves. Through HPLC analysis, chlorogenic acid and luteolin-7-O-glucoside were identified as predominant constituents in the ethanol extract. Their anti-inflammatory effects were evaluated using murine macrophage (RAW 264.7 cells) and human lung carcinoma cells (NCI-H292 & A549). The ethanol extract significantly (p<0.01) inhibited the production of nitrite, interleukin-6 (IL-6), and prostaglandin E2 (PGE2) induced by lipopolysaccharide (LPS) in RAW 264.7 cells. Furthermore, the ethanol extract suppressed the expression of cyclooxygenase-2 (COX-2) and inducible NO synthase (iNOS) proteins in RAW 264.7 cells stimulated with LPS. In NCI-H292 and A549 cells, treatment with the ethanol extract significantly (p<0.05) decreased levels of pro-inflammatory cytokines IL-6 and IL-8 induced by IL-1β. The phosphorylation of ERK rather than JNK in the mitogen-activated protein kinase signaling pathway was observed to be a more important mediator in the down-regulation of pro-inflammatory cytokines in NCI-H292 cells. These findings suggest that the ethanol extract of Platycodon grandiflorum leaves containing luteolin-7-O-glucoside exhibits promising anti-inflammatory properties.

주제어

표/그림 (8)

그림 Fig. 1. HPLC chromatograms of standard, ethanol extract and standard+ethanol extract at 348 nm in spike tests. (A) chlorogenic acid and (B) luteolin-7-O-glucoside.
표 Table 1. Linearity, precision, accuracy and limit of HPLC analysis method for chlorogenic acid and luteolin-7-O-glucoside
그림 Fig. 2. Effects of the ethanol extracts on the cell viabilities. (A) RAW 264.7 cells were treated with the indicated concentrations of the extracts of PGL (Platycodon grandiflorum leaves) and stimulated with LPS (100 ng/mL), and (B) NCI-H292 & (C) A549 cells treated with the extract of PGL and stimulated with IL-1β (10 ng/mL). Cell viabilities were analyzed by EZ-Cytox and data are presented as mean±S.D. (n=3). ES-PGL: ethanol extract of PGL harvested in spring, EF-PGL: ethanol extract of PGL harvested in fall.
표 Table 2. Contents of chlorogenic acid and luteolin-7-O-glucoside in extracts of Platycodon grandiflorum leaves evaluated from HPLC analysis
그림 Fig. 3. Effects of ethanol extracts on the levels of nitrite, IL-6 and PGE2 in LPS-stimulated RAW 264.7 cells. RAW 264.7 cells were treated with the indicated concentrations of the extracts of PGL (Platycodon grandiflorum leaves) and stimulated with LPS (100 ng/mL). (A) Nitrite, (B) IL-6 and (C) PEG2 were analyzed by ELISA kits and data are presented as mean±SD (n=3). Ethanol extract treatment groups were compared with LPS-stimulated group without extract treatment by Dunnet t-test (*p<0.05, **p<0.01). ES-PGL: ethanol extract of PGL harvested in spring, EF-PGL: ethanol extract of PGL harvested in fall.
그림 Fig. 4. Effects of ethanol extracts on the protein expressions of COX-2 and iNOS in LPS-stimulated RAW 264.7 cells. RAW 264.7 cells were treated with the indicated concentrations of the extracts of PGL (Platycodon grandiflorum leaves) and stimulated with LPS (100 ng/mL). Western blot analysis of cell lysates showed the protein expressions of COX-2 and iNOS. (A) Representative image of western blot and (B) the relative expression levels of COX-2 protein were normalized as an internal protein of β-actin. Data are presented as mean±S.D. (n=3). Ethanol extract treatment groups were compared with LPS-stimulated group without extract treatment by Dunnet t-test (*p<0.05, **p<0.01). ES-PGL: ethanol extract of PGL harvested in spring, EF-PGL: ethanol extract of PGL harvested in fall.
그림 Fig. 5. Inhibitory effects of ethanol extracts on cytokines productions. NCI-H292 & A549 cells were treated with the extracts of PGL (Platycodon grandiflorum leaves) and stimulated with IL-1β (10 ng/mL). The levels of (A) IL-6 & (B) IL-8 in NCI-H292 cells, and (C) IL-6 & (D) IL-8 in A549 cells were evaluated using ELISA kits and data are presented as mean±S.D. (n=3). Data are Ethanol extract treatment groups were compared with IL-1β-stimulated group without extract treatment by Dunnet t-test (*p<0.05; **p<0.01). ES-PGL: ethanol extract of PGL harvested in spring, EF-PGL: ethanol extract of PGL harvested in fall.
그림 Fig. 6. Inhibitory effects of ethanol extracts on the phosphorylation of MAPK signaling proteins. NCI-H292 were treated with the extracts of PGL (Platycodon grandiflorum leaves) and stimulated with IL-1β (10 ng/mL). Western blot analysis of cell lysates showed the expression levels of the phosphorylated or unphosphorylated MAPK signaling proteins. Representative images of western blots of (A) p-ERK & ERK and (C) p-JUK & JNK are presented and phosphorylated signals of (B) ERK & (D) JNK were normalized to unphosphorylated forms and expressed as a ratio to IL-1β-stimulated group without extract. Data are presented as mean±S.D. (n=3). Ethanol extract treatment groups were compared with IL-1β -stimulated group without extract by Dunnet t-test (*p<0.05, **p<0.01). ES-PGL, ethanol extract of PGL harvested in spring; EF-PGL, ethanol extract of PGL harvested in fall.

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용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

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