[논문]산수유의 채취 부위에 따른 기능 성분 분석과 항산화 및 항염증 효과에 관한 연구

황보현; 정지숙; 김민영; 지선영; 윤선혜; 김태희; 김성옥; 최영현

doi:10.5352/jls.2021.31.3.287

산수유의 채취 부위에 따른 기능 성분 분석과 항산화 및 항염증 효과에 관한 연구
A Study on Antioxidant and Anti-inflammatory Effects Based on Analysis of Functional Components of Cornus officinalis Siebold & Zucc. 원문보기

생명과학회지 = Journal of life science, v.31 no.3, 2021년, pp.287 - 297

황보현 (동의대학교 항노화연구소) , 정지숙 (구례군농업기술센터 야생화연구소) , 김민영 (동의대학교 항노화연구소) , 지선영 (동의대학교 항노화연구소) , 윤선혜 ((주)네이처텍) , 김태희 (함소아제약 중앙연구소) , 김성옥 (경성대학교 식품영양학과) , 최영현 (동의대학교 항노화연구소)

초록
AI-Helper

산수유 나무는 그 열매인 산수유와 함께 동아시아의 많은 국가에서 식용 및 약용으로 오랫동안 널리 사용되고 있다. 선행 연구들에 의하면 산수유 나무 추출물 및 구성 성분은 다양한 약리학적 잠재력을 가지고 있음에도 불구하고 부위별 추출물에 대한 체계적인 성분 분석은 아직 충분하지 않은 실정이다. 본 연구에서는 산수유 나무의 각 부위(청산수유, 적산수유, 씨앗, 줄기, 잎, 건피)의 유효 성분 함량을 분석하여 항산화 활성을 검증하였다. 본 연구의 결과에 따르면 조사된 3가지 지표 성분 중 morroniside은 청산수유와 적산수유에 가장 많이 함유하고 있었으며, loganin은 청산수유와 씨앗에 많았다. 잎, 적산수유, 건피의 함량은 비슷한 수준이었다. Cornin은 잎에 가장 많았으며, 청산수유, 적산수유 건피 순이었다. 또한 총 폴리페놀 함량은 적산수유, 청산수유, 잎 순으로 높았으며, 총 플라보노이드 함량은 잎이 가장 높았다. 그리고, DPPH radical 소거능은 잎, 씨앗, 적산수유 추출물 순으로 높은 소거 활성을 나타냈다. 따라서 항산화 활성이 가장 높은 잎 추출물의 항염증 효능을 조사하였으며, 이를 위하여 LPS로 자극된 RAW 264.7 대식세포에서 NO와 같은 염증성 매개 인자와 IL-1β와 IL-6를 포함한 염증성 사이토카인의 생성에 미치는 잎 추출물의 영향을 평가하였다. 본 연구의 결과에 의하면 세포독성이 없는 조건에서 잎 추출물은 LPS에 의하여 증가된 iNOS의 발현과 NO의 생성을 유의적으로 억제하였다. 또한 잎 추출물은 LPS에 의한 IL-1β 및 IL-6의 생성 및 발현을 현저하게 감소시켰다. 따라서 산수유 나무의 잎 추출물은 항산화제와 항염증제의 개발을 위한 천연 재료로서 잠재력이 있음을 시사하며, 관련 기전 연구가 추가로 진행되어야 할 것이다.

Abstract ▼ AI-Helper

Cornus officinalis Siebold & Zucc. is traditionally used as an edible and medicinal plant in many countries in East Asia. Previous studies have shown the pharmacological potential of extracts and components of C. officinalis, but comparative analysis of the composition of the leaf, stem, and fruit extracts has been insufficient to date. In the present study, the content of active antioxidant and anti-inflammatory ingredients was verified in different C. officinalis parts (under-ripe sansuyu, ripe sansuyu, seed, leaf, stem, and dried sansuyu). One active component, morroniside, was high in fruit (under-ripe and ripe sansuyu), while loganin was high in fruit (under-ripe sansuyu) and cornin was high in seeds. Total polyphenol contents were highest in fruit (ripe sansuyu) and flavonoids were highest in leaves. DPPH radical scavenging activity was highest in leaves, followed by seeds and then ripe sansuyu extract. The anti-inflammatory efficacy of leaf extracts of C. officinalis (LCO) was further investigated by measuring their effects on levels of nitric oxide (NO) and the pro-inflammatory cytokines interleukin (IL)-1β and IL-6 in RAW 264.7 macrophages. Non-cytotoxic concentrations of LCO effectively decreased the lipopolysaccharide (LPS)-induced expression of inducible NO synthase, resulting in decreased NO production. LCO also significantly suppressed LPS-induced production and expression of IL-1β and IL-6. Taken together, the present findings suggest that C. officinalis leaves have potential as natural materials for the development of antioxidant and anti-inflammatory agents.

주제어

표/그림 (10)

표 Table 1. HPLC/DAD operating condition for morroniside, loganin, cornin and extracts from different parts of C. officinalis
표 Table 2. List of antibodies used for western blot analysis in the present study
그림 Fig. 1. Total polyphenol contents of extract from different parts of C. officinalis. Value are mean ± SD (n=3). Different letters (a-f) above the bars are significant differences at p<0.05 by Duncan's multiple range test.
그림 Fig. 2. Total flavonoid contents of extract from different parts of C. officinalis. Value are mean ± SD (n=3). Different letters (a-d) above the bars are significant differences at p<0.05 by Duncan's multiple range test.
그림 Fig. 3. DPPH free radical scavenging ability of extracts from different parts of C. fructus. Value are mean ± SD (n=3). Different letters (a-f) above the bars are significant differences at p<0.05 by Duncan's multiple range test.
표 Table 3. Active ingredients of morroniside, loganin and cornin of extracts from different parts of C. officinalis
그림 Fig. 4. Effects of the extracts from different parts of C. officinalis on NO production and cell viability in LPS-stimulated RAW 264.7 cells. The cells were pre-incubated with or without the indicated extracts from different parts of C. officinalis for 1 hr, and then treated with or without LPS (100 ng/ml) for 24 hr. (A) The nitrite concentration in culture media was evaluated by Greiss reaction. (B) Cell viability was determined by an MTT assay. Results are expressed as the percentage of surviving cells over control cells. Each data point represents mean ± SD of three independent experiments.
그림 Fig. 5. Effects of leaf extracts of C. officinalis (LCO) on cell viability in RAW 264.7 cells. (A) The cells were treated with various concentrations of LCO extracts for 24 hr. (B) The cells were pretreated with the indicated concentrations of LCO for 1 hr prior to treatment with 100 ng/ml LPS for 24 hr. Cell viability was assessed by MTT assay. The results are expressed as means ± SD of three independent experiments duplicate in each run.
그림 Fig. 6. Inhibition of NO production and iNOS expression by leaf extracts of C. officinalis (LCO) in RAW 264.7 cells. The cells were exposed to the indicated concentrations of LCO 1 hr prior treatment of LPS (100 ng/ml) for 24 hr. (A) Amounts of NO production in culture media were determined by Greiss reaction. (B). Total proteins were isolated and subjected to Western blot analysis with specific antibody against iNOS. Experiments were repeated three times and similar results were obtained. Actin was used as a control for equal loading. iNOS/actin ratio was calculated and analyzed. All experiments were repeated three times (^#p<0.05 in comparison with the control group; * p<0.05 in comparison with the LPS group).
그림 Fig. 7. Inhibition of IL-1β and IL-6 secretion and expression by leaf extracts of C. officinalis (LCO) in RAW 264.7 cells. The cells were exposed to the indicated concentrations of LCO 1 hr prior treatment of LPS (100 ng/ml) for 24 hr. (A and C) The IL-1β (A) and IL-6 (C) levels in medium were evaluated using commercial IL-1β and IL-6 cytokine ELISA kits. (B and D) Total proteins were isolated and subjected to Western blot analysis with specific antibodies against IL-1β and IL-6. Experiments were repeated three times and similar results were obtained. Actin was used as a control for equal loading. IL-1β and IL-6/actin ratio were calculated and analyzed. All experiments were repeated three times (^#p<0.05 in comparison with the control group; * p<0.05 in comparison with the LPS group).

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