Michael addition acceptor 그룹을 가지고 있는 phytochemicals의 toll-like receptor 신호전달체계 조절을 통한 항염증 효과

윤형선(Hyung-Sun Youn) 한국식품과학회 2009 한국식품과학회지 Vol.41 No.5

TLRs는 여러 병원균들이 가지고 있는 PAMPs를 인식해서, 선천성 면역 반응을 유도하는 중요한 역할을 한다. TLR4의 이합체 형성은 신호전달 체계의 활성화와 뒤이어 발생하는 선천성 면역 반응을 유도하기 위해서 최초로 일어나는 반응으로 알려져 있다. 우리가 먹는 식품 중에는 항염증 효과가 있다고 널리 알려져 있는 phytochemicals이 포함되어 있다. 특히 α,β-unsaturated carbonyl group을 가지고 있는 curcumin, 6-shogaol, 그리고 cinnamaldehyde는 Michael addition 반응에 의해서 LPS에 의해서 유도된 TLR4의 이합체 형성을 억제시켜, 전사요소 NF-κB와 IRF3 활성화 및 그것들에 의해서 조절되는 타깃 유전자들을 억제시킨다. 이러한 결과는 α,β-unsaturated carbonyl group을 가지고 있는 curcumin, 6-shogaol, 그리고 cinnamaldehyde의 항염증 효능에 대한 새로운 기전을 설명해 주는 것이라 할 수 있겠다. Toll-like receptors (TLRs) play a critical role in the induction of innate immune responses that are essential for host defense against invading microbial pathogens. In general, TLRs have two major downstream signaling pathways, namely MyD88- and TRIF-dependent pathways, leading to the activation of nuclear factor-κB (NF-κB) and interferon regulatory factor 3 (IRF3) and the expression of inflammatory mediators. TLR4 dimerization is required for the activation of downstream signaling pathways and may be one of the first lines of regulation in activating TLR-mediated signaling pathways. In this paper, the molecular targets of curcumin, 6-shogaol, and cinnamaldehyde in TLR signaling pathways will be discussed. Curcumin, 6-shogaol, and cinnamaldehyde with α,β-unsaturated carbonyl groups inhibit the dimerization of TLR4 induced by lipopolysaccharide, resulting in the downregulation of NF-κB and IRF3. These results suggest that phytochemicals with the structural motif conferring Michael addition inhibit TLR4 dimerization, suggesting a novel mechanism for the anti-inflammatory activity of phytochemicals.

Toll-like receptors 신호전달체계 조절을 통한 resveratrol, (-)-epigallocatechin-3-gallate, curcumin의 항염증 효과

윤형선(Hyung-Sun Youn) 한국식품과학회 2007 한국식품과학회지 Vol.39 No.5

Toll-like receptors (TLRs) induce innate immune responses that are essential for host defenses against invading microbial pathogens, thus leading to the activation of adaptive immune responses. In general, TLRs have two major downstream signaling pathways: the MyD88- and TRIF-dependent pathways, which lead to the activation of NF-_κand IRF3. Numerous studies have demonstrated that certain phytochemicals possessing anti-inflammatory effects inhibit NF-_κB activation induced by pro-inflammatory stimuli, including lipopolysaccharides and TNFα. However, the direct molecular targets for such anti-inflammatory phytochemicals have not been fully identified. Identifying the direct targets of phytochemicals within the TLR pathways is important because the activation of TLRs by pro-inflammatory stimuli can induce inflammatory responses that are the key etiological conditions in the development of many chronic inflammatory diseases. In this paper, we discuss the molecular targets of resveratrol, (-)-epigallocatechin-3-gallate (EGCG), and curcumin in the TLR signaling pathways. Resveratrol specifically inhibited the TRIF pathway in TLR3 and TLR4 signaling, by targetting TBK1 and RIP1 in the TRIF complex. Furthermore, EGCG suppressed the activation of IRF3 by targetting TBK1 in the TRIFdependent signaling pathways. In contrast, the molecular target of curcumin within the TLR signaling pathways is the receptor itself, in addition to IKKβ. Together, certain dietary phytochemicals can modulate TLR-derived signaling and inflammatory target gene expression, and in turn, alter susceptibility to microbial infection and chronic inflammatory diseases.

Toll-like receptor 2, 3, 4의 신호전달체계 조절을 통한 curcumin의 항암ㆍ항염증 효과

윤형선(Hyung-Sun Youn),강순아(Soon Ah Kang),Daniel Hwang 한국식품과학회 2007 한국식품과학회지 Vol.39 No.2

TLRs는 병원균이 숙주의 몸속에 들어 왔을 때, 병원균들이 기지고 있는 독특한 구조를 인식하여 선천성 면역반응과 뒤이어 후천성 면역반응을 유도하는 중요한 역할을 한다. 우리는 이번 실험을 통하여 curcumin이 선행연구에서 밝혀낸 TLR4 뿐만 아니라 TLR2와 TLR6 그리고 TLR3를 또한 분자학적인 타깃으로 할 수 있다는 것을 알아내었다. Curcumin이 MALP-2(TLR2,6 agonist)에 의해서 유두된 IRAK-1 degradation을 억제시켰다. 이러한 결과는 curcumin의 분자학적인 타깃이 IRAK-1위에 놓여 있으며, TLR2와 TLR6가 될 것이라는 가능성을 제시해 준다고 할 수 있다. 또한 curcumin은 viral 자극제인 poly[I:C](TLR3 agonist)에 의해서 유도된 IRF3나 NF-κB 활성화를 억제하였지만, TRIF에 의해서 유도된 IRF3 활성화는 억제시키지를 못하였다. 이러한 결과 또한 TLR3 자체가 curcumin의 분자학적인 타깃이라는 가능성을 제시해 준다고 할 수 있겠다. 이러한 결과를 종합해 볼때, curcumin의 분자학적인 타깃이 IKKβ 이외에 모든 TLRs가 될 수 있다는 가능성을 제시해 준다고 할 수 있겠다. 이러한 결과는 curcumin이 그람음성균 뿐만이 아니라 바이러스나 박테리아 등 여러 병원균들로부터 유도되는 염증반응이나 민성적인 질병들을 조절할 수 있다는 것을 보여주는 결과라 할 수 있겠다. Toll-like receptors induce innate immune responses recognizing conserved microbial structural molecules that are known as pathogen-associated molecular patterns (PAMPs). Ligand-induced homotypic oligomerization was found to proceed in LPS-induced activation of TLR4 signaling pathways. TLR2 is known to heterodimerize with TLR1 or TLR6 and recognize diacyl- or triacyl-lipopeptide, respectively. These results suggest that ligand-induced receptor dimerization of TLR4 and TLR2 is required for the activation of downstream signaling pathways. Therefore, receptor dimerization may be one of the first lines of regulation in the activation of TLR-mediated signaling pathways and induction of subsequent innate and adaptive immune responses. Here, we report biochemical evidence that curcumin from the plant Curcuma Zanga inhibits activation of NF-κB, expression of COX-2, and dimerization of TLRs induced by TLR2, TLR3 and TLR4 agonists. These results imply that curcumin can modulate the activation of TLRs and subsequent immune/inflammatory responses induced by microbial pathogens.

광귤나무(Citrus aurantium) 열매에서 분리된 쿠마린의 iNOS 억제효과

김범선(Beom sun Kim),정의길(Eui Gil Jung),윤형선(Hyung Sun Youn),황경아(Kyung A Hwang),한만덕(Man Deuk Han),유연호(Yon ho Yu),남궁우(Kung Woo Nam) 대한약학회 2017 약학회지 Vol.61 No.5

Citrus fruits have been used as an edible fruit, and also as a traditional medicine, since ancient times. A crude methanol extract of Citrus aurantium was shown to suppress inducible proinflammatory cytokines, but its constituents were not known. In this study, we isolated four furanocoumarins, bergapten (1), imperatorin (2), isoimperatorin(3), and phellopterin (4), and one 7-geranyloxycoumarin, auraptene (5), from a methanol extract of dried Citrus aurantium fruit. The structures of these compounds were elucidated by extensive application of spectroscopic techniques, including 2D NMR spectroscopy and mass spectrometry. All of the isolated compounds were evaluated for their ability to inhibit the production of proinflammatory cytokine inducible nitric oxide synthase (iNOS) in lipopolysaccharide-activated RAW264.7 cells. Compounds 2−5 significantly suppressed iNOS production but compound 1 had no iNOS inhibitory effects. These results indicate that at least three furanocoumarins and one 7-geranyloxycoumarin are involved in the suppressive effects of citrus fruits against the production of inducible inflammatory cytokines.

일반대기 중 극미세입자와 중금속 농도에 관한 연구

조태진,전혜리,윤형선,이미영,손부순,Cho, Tea-Jin,Jeon, Hye-Li,Youn, Hyung-Sun,Lee, Mi-Young,Son, Bu-Soon 한국환경보건학회 2010 한국환경보건학회지 Vol.36 No.6

This study evaluated the distribution of the concentrations of nano-particles and heavy metals (08-Pb, Cr, Zn, As, Fe, 09-Pb, Cr, Zn, Cu, Ni, Mn) in Seoul, Chungnam A and Gwangyang from August to December, in 2008 5 times each in the Seoul area, 5 times in and Chungnam A area and from August to November, in 2009 14 times in the Chungnam A area, 8 times in the Gwangyang area. The examined results showed high concentration level from $PM_1$ through $PM_{0.1}$ in all three areas. These results were obtained the concentration of particles by diameter and statistically significant in Stage5 (1.0-0.56 ${\mu}m$) from the result of conducting Kruskal-Wallis H test (p < 0.05). In the case of the heavy metal concentration included in 0.10-0.056 ${\mu}m$, 0.056 ${\mu}m$, the lead concentration of Chungnam Asan area was 6.49 ng/$m^3$ and 9.93 ng/$m^3$, which was higher than 3.05 ng/$m^3$ and 4.22 ng/$m^3$ of Seoul, respectively. The concentration of iron in Seoul was 9.28 ng/$m^3$ and 13.24 ng/$m^3$, that appeared higher than 2.38 ng/$m^3$ and 3.23 ng/$m^3$ of Chungnam A area, respectively. The concentration level was similar to other metals except lead and iron in Chungnam A area and Seoul. From the concentration of heavy metal included in 0.10-0.056 ${\mu}m$, 0.056 ${\mu}m$, the lead concentration of Chungnam A area was 0.31 ng/$m^3$ and 0.12 ng/$m^3$ while Gwangyang was 0.28 ng/$m^3$, 0.06 ng/$m^3$. Thus Chungnam A area showed higher lead concentration than Gwangyang. The manganese concentration of Chungnam A area was 0.12 ng/$m^3$ and 0.03 ng/$m^3$ while Gwangyang was 0.21 ng/$m^3$ and 0.08 ng/$m^3$. Therefore, the concentration of Gwangyang appeared higher than that of Chunnam A area. These two metals showed statistically significant in 0.056 ${\mu}m$ (p < 0.05, p < 0.01). Among the concentration of heavy metal in all regions, the result demonstrated that the order of higher concentration is arsenic > iron > zinc > chrome > lead > nickel > copper > manganese.

Ovalbumin에 의해서 유도된 inducible nitric oxide synthase 발현에 대한 phenethyl isothiocyanate의 억제효과

신화정(Hwa-Jeong Shin),윤형선(Hyung-Sun Youn) 한국식품과학회 2012 한국식품과학회지 Vol.44 No.6

이번 실험을 통하여 PEIC가 OVA에 의해 유도된 NF-κB 활성과 iNOS, COX-2 발현에 어떠한 영향을 미치는지 알아 보았다. PEIC는 OVA에 의해 유도된 NF-κB 활성을 억제시켰다. 또한 PEIC는 OVA에 의해 유도된 iNOS의 발현도 억제시켰다. 하지만 PEIC는 OVA에 의해 유도된 COX-2 발현은 억제시키지 못하였다. 이러한 결과는 iNOS와 COX-2가 서로 다른 메커니즘에 의해 조절된다는 것을 암시한다. 또한 PEIC는 알러지와 같은 만성적인 질병들을 조절할 수 있는 치료제 개발 및 백신 제조에 중요한 역할을 할 것으로 기대한다. Egg allergies have been reported as one of the most prevalent food hypersensitivities in the pediatric population. One of the major egg allergens is ovalbumin (OVA), which is the major protein in the egg whites. Phenethyl isothiocyanate (PEIC) from cruciferous vegetables has an effect on anti-inflammatory therapy. In the present report, we show that PEIC inhibits the nuclear factor-κB (NF-κB) activation induced by OVA. PEIC also inhibits the OVA-induced inducible nitric oxide synthase (iNOS) expression and nitrite production. However, PEIC did not suppress the cyclooxygenase- 2 (COX-2) expression induced by OVA. These results suggest that PEIC has the specific mechanism for anti-inflammatory responses and efficient anti-allergic activities.

Isoliquiritigenin의 toll-like receptor agonists에 의해서 유도된 NF-κB 활성화와 cyclooxygenase-2 발현 억제

박세정(Se-Jeong Park),양승주(Seung-Ju Yang),윤형선(Hyung-Sun Youn) 한국식품과학회 2009 한국식품과학회지 Vol.41 No.2

선천성 면역 반응을 위해 중요한 역할을 하는 TLRs가 외부 병원성 물질에 자극을 받게 되면 NF-κB를 활성화시키며, 그 결과로 염증을 유도하는 COX와 같은 유전자를 발현한다. 이번 연구에서, 옛날부터 지금까지 전통적인 약재로써 질병 치료에 다양하게 쓰이고 있는 감초의 뿌리에서 추출한 성분 중의 하나인 ILG가 NF-κB활성과 COX 발현을 어떻게 조절하여 항염증 효과를 가지고 있는지 알아보았다. ILG는 TLR agonists인 MALP-2, Poly[I:C], 그리고 LPS에 의해 유도된 NF-κB 활성화와 COX-2 발현을 억제시켰다. 또한 ILG는 리간드(ligand)에 독립적인 TLRs signaling downstream molecules인 MyD88, IKKβ, 그리고 p65에 의해서 유도된 NF-κB 활성을 억제시켰다. 이러한 결과는 한약재로서 많이 이용되는 감초가 단지 한약의 쓴맛을 줄이기 위함이 아니라 TLRs 신호전달 체계를 조절하여 항염증 효과를 가지고 있다는 것을 보여주는 것이라 할 수 있겠다. Toll-like receptors(TLRs) are pattern recognition receptors(PRRs) that recognize pathogen-associated molecular patterns(PAMPs) and regulate the activation of innate immunity. All TLR signaling pathways culminate in the activation of NF-κB, leading to the induction of inflammatory gene products such as COX-2. Licorice (Glycyrrhiza uralensis) has been used for centuries as an herbal medicine. Isoliquiritigenin(ILG), a simple chalcone-type flavonoid, is an active component present in licorice and has been used to treat many chronic diseases. However, the mechanism as to how ILG mediates health effects is still largely unknown. In the present report, we present biochemical evidence that ILG inhibits the NF-κB activation induced by TLR agonists and the overexpression of downstream signaling components of TLRs, MyD88, IKKβ, and p65. ILG also inhibits TLR agonists-induced COX-2 expression. These results suggest that anti-inflammatory effects of ILG are caused by modulation of the immune responses regulated by TLR signaling pathways.

Parthenolide Suppresses the Expression of Cyclooxygenase-2 and Inducible Nitric Oxide Synthase Induced by Toll-Like Receptor 2 and 4 Agonists

A-Neum Lee,Se-Jeong Park,Sae-Mi Yun,Mi-Young Lee,Bu-Soon Son,Hyung-Sun Youn(윤형선) 대한의생명과학회 2010 Biomedical Science Letters Vol.16 No.1

Toll-like receptors (TLRs), which are pattern recognition receptors (PRRs), recognize pathogen-associated molecular patterns (PAMPs) and regulate the activation of innate immunity. All TLR signaling pathways culminate in the activation of NF-κB, leading to the induction of inflammatory gene products such as cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS). Parthenolide, a sesquiterpene lactone isolated from the herb feverfew (Tanacetum parthenium), has been used as folk remedies to treat many chronic diseases for many years. In the present report, we present biochemical evidence that parthenolide inhibits the NF-κB activation induced by TLR agonists and the overexpression of downstream signaling components of TLRs, MyD88, IKKβ, and p65. Parthenolide also inhibits TLR agonists-induced COX-2 and iNOS expression. These results suggest that parthenolide can modulate the immune responses regulated by TLR signaling pathways.

6-Shogaol의 Toll-like receptor 2, 3, 4 agonists에 의해서 유도된 cyclooxygenase-2 발현 억제

김점지(Jum-Ji Kim),안상일(Sang-Il Ahn),이전수(Jun-Su Lee),윤새미(Sae-Mi Yun),이미영(Mi-Young Lee),윤형선(Hyung-Sun Youn) 한국식품과학회 2008 한국식품과학회지 Vol.40 No.3

선천성 면역은 병원성균의 침입에 대항하기 위한 숙주의 최초 방어체계라 할 수 있다. 이러한 선천성 면역반응은 병원균들이 가지고 있는 독특한 구조를 인식하는 TLRs에 의해서 조절되어 진다고 알려져 있다. 숙주에 침입한 여러 병원성균들이 TLRs를 자극하며 이렇게 자극된 신호들은 아래로 전달되어 전사요소 NF-κB의 활성화를 유도하고 결국 COX-2와 같은 염증 유발인자를 유도하여 암이나 질병을 유발하게 된다. 우리는 이번 연구를 통하여 생강 추출물중의 하나인 6-shogaol이 어떻게 NF-κB 활성화나 COX-2 발현을 조절하여 항염증 효과를 가지고 있는지를 알아보았다. 6-shogaol은 TLR2, TLR3, TLR4 agonists에 의해서 유도된 NF-κB 활성화와 COX-2 발현을 억제하였다. 이러한 결과는 6-shogaol이 여러 병원균들로부터 유도되는 염증반응이나 만성적인 질병들을 조절할 수 있다는 중요한 결과를 보여주는 것이라 할 수 있다. Ginger is widely used as a traditional herbal medicine. Both ginger and its extracts have been used to treat many chronic inflammatory conditions via the inhibition of nuclear factor-kappa B (NF-κB) activation, which results in the suppression of cyclooxygenase-2 (COX-2) expression. However, the mechanisms as to how ginger extracts mediate their health effects are largely unknown. Toll-like receptors (TLRs) trigger anti-microbial innate immune responses, recognizing conserved microbial structural molecules that are known as pathogen-associated molecular patterns. All TLR signaling pathways culminate in the activation of NF-κB. The activation of NF-κB leads to the induction of inflammatory gene products, including cytokines and COX-2. This study reports the biochemical evidence that 6-shogaol, an active compound in ginger, inhibits NF-κB activation and COX-2 expression induced by TLR2, TLR3, and TLR4 agonists. Furthermore, 6-shogaol inhibited NF-κB activation induced by the following downstream signaling components of the TLRs: MyD88, IKKβ, and p65. These results imply that ginger can modulate immune responses that could potentially modify the risk of many chronic inflammatory diseases.

Curcumin은 ovalbumin에 의해서 유도된 inducible nitric oxide synthase 억제

김지수(Ji Soo Kim),안희진(Hee Jin Ahn),신화정(Hwa-Jeong Shin),구교정(Gyo-Jeong Gu),엄상훈(Sang Hoon Eum),이청호(Chung Ho Lee),민인순(In Soon Min),윤형선(Hyung-Sun Youn) 한국식품과학회 2012 한국식품과학회지 Vol.44 No.4

본 연구에서 curcumin이 계란 알러젠 중의 하나인 OVA에 의해서 유도된 NF-κB 활성화 및 iNOS 발현에 어떤 영향을 미치는지 알아보았다. Curcumin은 OVA에 의해서 유도된 NF-κB 활성화와 iNOS 발현을 억제시켰다. 이러한 결과는 curcumin이 계란 알러젠인 OVA에 의해서 유도된 NF-κB의 활성화와 iNOS의 발현을 억제하여 염증반응이나 알러지와 같은 만성적인 질병들을 조절할 수 있다는 것을 보여주는 중요한 결과라 사료된다. 이러한 연구는 추후 알러지 작용기전 규명 및 알러지 치료제 개발에 중요한 역할을 할 것으로 기대한다. Egg allergy has been reported as the most prevalent food hypersensitivity among children. One of the major egg allergens is ovalbumin (OVA). OVA is the major protein in the egg white, comprising 54% of its total protein content. Curcumin isolated from Curcuma longa has been used as folk remedies in order to treat many chronic diseases for many years. In the present report, we present biochemical evidence that curcumin inhibits the NF-κB activation induced by OVA. Curcumin also inhibits OVA-induced iNOS expression and nitrite production. These data suggest new approaches for the development of efficient anti-allergic strategies.