[논문]감초 신품종 및 약전 수재감초의 면역조절 효과 비교 연구

강윤미; 김원남; 진종식; 이종현; 장재기; 이정훈; 안효진

doi:10.6116/kjh.2020.35.6.11.

[국내논문] 감초 신품종 및 약전 수재감초의 면역조절 효과 비교 연구
The Comparative Study of Immunomodulatory Effect by Glycyrrhiza New Varieties and Official Compendia 원문보기

大韓本草學會誌 = The Korea journal of herbology, v.35 no.6, 2020년, pp.11 - 19

강윤미 (상지대학교 한의과대학 약리학교실) , 김원남 (세명대학교 한의과대학 본초방제학교실) , 진종식 (전북대학교 환경생명자원대학 한약자원학과) , 이종현 (동덕여자대학교 약학대학 약학과) , 장재기 (농촌진흥청 국립원예특작과학원 약용작물과) , 이정훈 (농촌진흥청 국립원예특작과학원 약용작물과) , 안효진 (상지대학교 한의과대학 약리학교실)

Abstract ▼ AI-Helper

Objective : The genus Glycyrrhiza has been used in food and traditional herbal medicine. Glycyrrhiza new varieties Wongam and Sinwongam have been developed by Korea Rural Development Administration and investigated to register on Korean Pharmacopoeia of the Ministry of Food and Drug Safety. The aim of this study is to investigate the immunomodulatory effect of Wongam and Sinwongam comparing with listed Glycyrrhiza species (Glycyrrhiza uralensis Fischer and G. glabra Linne) for evaluations about pharmacological effect of Glycyrrhiza new varieties. Methods : We studied the immunomodulatory effect of Wongam and Sinwongam compared with G. uralensis and G. glabra using THP-1 cell in vitro model. The cells were treated with phorbol 12-myristate 13-acetate (PMA) for differentiation and stimulated with lipopolysaccharides (LPS) to induce immune activation. We analyzed and compared the effects Glycyrrhiza new varieties and listed Glycyrrhiza species using nitric oxide (NO) assay, western blot, and reverse transcription-quantitative polymerase chain reaction analysis. 1) Results : Wongam and Sinwongam showed no cytotoxicity in THP-1 cells. Wongam and Sinwongam, and listed Glycyrrhiza species increased NO production, and cyclooxygenase (COX)-2 expression with or without LPS in differentiated THP-1 macrophages. Furthermore, Wongam and Sinwongam and listed Glycyrrhiza species upregulated the mRNA expressions of T helper type 1 (Th 1)-associated cytokines in LPS-stimulated THP-1 macrophages. Conclusion : These results indicated that Wongam and Sinwongam would have effect of enhancing immune response through the increase of NO and COX-2 expression, and activate Th1-associated cytokines. The findings of this study suggest the wide applicability of Glycyrrhiza new varieties.

Keyword

표/그림 (5)

그림 Figure 1. Effects of four Glycyrrhiza species samples on cell viability on THP-1 cells. Cells were treated with different concentrations of Glycyrrhiza species samples for 24 h and their viability was determined using MTS assay. The data shown represent mean ± S.D. of three independent experiments. *p < 0.05, **p < 0.01 and ***p < 0.001 vs non-treated cells.
그림 Figure 2. Effects of four Glycyrrhiza species samples on NO production in THP-1 macrophages. (A) THP-1 cells differentiated by PMA were incubated with Glycyrrhiza species samples for 48 h. (B) THP-1 cells differentiated by PMA were incubated with Glycyrrhiza species samples with LPS (10 ㎍/㎖) for 48 h. NO levels were determined with Griess reagent. The data shown represent mean ± S.D. of three independent experiments. ^###p < 0.001 vs the control group, **p < 0.01, and ***p < 0.001 vs (A) control group or (B) LPS-stimulated cells.
그림 Figure 3. Effects of four Glycyrrhiza species samples on COX-2 protein expression in THP-1 macrophages. (A) THP-1 cells differentiated by PMA were incubated with Glycyrrhiza species samples or samples plus LPS (10 ㎍/㎖) for 24 h or (B) 48 h. The protein level of COX-2 was determined by western blot analysis using specific antibodies. Densitometric analysis was performed using Bio-Rad Quantity One software. The data shown represent mean ± S.D. of three independent experiments. ^$$$p < 0.001 vs the non-treated group, ^###p < 0.001 vs the differentiation group, **p < 0.01, and *** p < 0.001 vs LPS-stimulated cells.
그림 Figure 4. Effects of four Glycyrrhiza species samples on mRNA expressions of TNF-α, and IL-6 in differentiated THP-1 macrophages. THP-1 cells differentiated by PMA were incubated with Glycyrrhiza species samples with LPS (10 ㎍/㎖) for 6 h. Total RNA prepared form the dorsal tissue, and the levels of (A) TNF-α, and (B) IL-6 were determined by qRT-PCR. The data shown represent mean ± S.D. of three independent experiments. ^###p < 0.001 vs the control group,**p < 0.01, and ***p < 0.001 vs LPS-stimulated cells.
그림 Figure 5. Effects of four Glycyrrhiza species samples on mRNA expressions of IL-2, IL-12, and IL-10 in differentiated THP-1 macrophages. THP-1 cells differentiated by PMA were incubated with Glycyrrhiza species samples with LPS (10 ㎍/㎖) for 6 h. Total RNA prepared form the dorsal tissue, and the levels of (A) IL-2, (B) IL-12, and (C) IL-10 were determined by qRT-PCR. The data shown represent mean ± S.D. of three independent experiments. ^###p < 0.001 vs the control group, *p < 0.05, **p < 0.01, and *** p < 0.001 vs LPS-stimulated cells.

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제안 방법

To investigate whether Glycyrrhiza species samples were cytotoxic to the THP-1 cells, the MTT assay was performed. The extracts of Wongam, Sinwongam, G.

대상 데이터

TNF-α, IL-6, IL-2, IL-10, IL-12, and GAPDH oligonucleotide primers were purchased from Bioneer Corporation (Daejeon, South Korea).

데이터처리

Statistically significant differences were compared using one-way analysis of variance and Dunnett’s post hoc test.

이론/모형

, Waltham, MA, USA) with SYBR Premix Ex Taq. Fold changes of gene expression were calculated using the comparative quantification cycle method. The Cq values of target genes TNF-α, IL-6, IL-2, IL-10, and IL-12 were normalized to that of GAPDH using the ABI Gene Express 2.
Effects of four Glycyrrhiza species samples on cell viability on THP-1 cells. Cells were treated with different concentrations of Glycyrrhiza species samples for 24 h and their viability was determined using MTS assay. The data shown represent mean ± S.

성능/효과

The decoction was filtered, lyophilized and stored at 4℃. The yields of the dried extract from the starting crude of Wongam, Sinwongam, G. uralensis and G. glabra were 8.8 %, 9.13 %, 6.53 % and 7.71 %, respectively. To prepare the sample for the in vitro experiment, the extract powder that resulted from the drying process was dissolved in distilled water.
In conclusion, this study showed that Glycyrrhiza new varieties Wongam and Sinwongam have an immunomodulatory effect and enhance immune function by regulating the NO, COX-2, and immune-associated cytokines derived from activated macrophages. Compared to listed Glycyrrhiza species, the higher NO level was induced by Wongam treatment in differentiated THP-1 cells, and the Sinwongam treatment activated immune responses increasing COX-2, and TNF-α expression in LPS-stimulated THP-1 macrophages.
Compared to listed Glycyrrhiza species, the higher NO level was induced by Wongam treatment in differentiated THP-1 cells, and the Sinwongam treatment activated immune responses increasing COX-2, and TNF-α expression in LPS-stimulated THP-1 macrophages. The findings of this study imply that Wongam and Sinwongam have an immunomodulation efficacy homogeneity with listed Glycyrrhiza species in the Korean Pharmacopoeia, thus suggest the wide applicability of Glycyrrhiza new varieties Wongam and Sinwongam.

후속연구

IL-10 is also known as an anti-inflammatory cytokine which regulates cytokinebalance and limits the overt inflammation preventing tissue damage³⁵⁾. As licorice has a well-documented anti-inflammatory property, further studies should investigate the effect of Wongam, Sinwongam on antiinflammatory action and more specific Th1 cell-mediated immune responses. These further researches on Glycyrrhiza species might extend the explanations of the immunomodulatory effect of Wongam and Sinwongam.

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