Modulation of Nrf2/ARE and Inflammatory Signaling Pathways by Hericium erinaceus Mycelia Extract

Kyong-Suk Jin,Ji-Young Park,Mi-Kyung Cho,Ji-Hyun Jang,Jae-Han Jeong,Seon Ok,Min-Ji Bak,Young-Sun Song,Myo-Jeong Kim,Chung-Won Cho,Woo-Sik Jeong 한국식품과학회 2009 Food Science and Biotechnology Vol.18 No.5

Hericium erinaceus is an edible mushroom used as a medicinal food in Asian countries. In this study, the chemopreventive effects of H. erinaceus mycelia hot water extract (HEW) were evaluated. HEW remarkably induced the luciferase activity of the antioxidant response element (ARE), located in the promoter region of phase 2 and antioxidant genes and regulated by nuclear factor E2-related factor 2 (Nrf2). The up-regulation of ARE activity by HEW corresponded with the induction of Nrf2 and the antioxidant enzyme, hemeoxygenase-1. The inhibition of cyclooxygenase-2 (COX-2) activity is a promising effective approach in cancer chemoprevention, and HEW prominently suppressed COX-2 protein expression in HepG2 cells. Furthermore, HEW showed anti-inflammatory activity by modulating inflammatory mediators such as nitric oxide (NO), inducible NO synthase, tumor necrosis factor-α, interleukin-1β, and the transcription factor, nuclear factor-κB, in lipopolysaccharide-stimulated RAW 264.7 cells. These results suggest that H. erinaceus possessed anti-tumor and antiinflammatory effects via the modulation of Nrf2/ARE and inflammatory signaling pathways, and may therefore have potential use as a natural chemopreventive agent.

Vitis amurensis Ruprecht root inhibited α-melanocyte stimulating hormone-induced melanogenesis in B16F10 cells

Kyong-Suk Jin,You Na Oh,Sook Kyung Hyun,Hyun Ju Kwon,Byung Woo Kim 한국영양학회 2014 Nutrition Research and Practice Vol.8 No.5

BACKGROUND/OBJECTIVES: The root of Vitis amurensis Ruprecht, a sort of wild-growing grape, has been used in oriental medicine for treatment of skin ailments; however, its dermatological activity is not sufficiently understood. The aim of this study was to investigate tyrosinase inhibitory and anti-melanogenic activities of V. amurensis Ruprecht root methanol extract (VARM) in B16F10 mouse melanoma cells and to attempt to isolate and identify the active compound issued from VARM. MATERIALS/METHODS: Anti-melanogenic activity of VARM was analyzed in α-melanocyte stimulating hormone (MSH)-stimulated B16F10 cells through evaluation of antioxidative activity as well as inhibited tyrosinase activity and melanin contents compared with those of kojic acid and arbutin. After anti-melanogenic analysis of VARM, serial fractionation, nuclear magnetic resonance (NMR), and thin layer chromatorgraphy (TLC) were applied for identification of active compounds contained in VARM. RESULTS: VARM significantly inhibited oxidative stress and tyrosinase activity and attenuated α-MSH-induced melanin production in B16F10 cells. For isolation of active compounds, VARM was fractionated using a series of organic solvents, including dichloromethane (CH₂Cl₂), ethyl acetate (EtOAc), and n-butanol (n-BuOH). Among fractions showing anti-melanogenic activity, the CH2Cl2 fraction induced the most potent attenuation of melanogenesis without cytotoxicity and the major compound in the CH₂Cl₂ fraction was identified as betulinic acid. Betulinic acid isolated from the CH₂C₂ fraction of VARM significantly attenuated α-MSH-induced melanogenesis in a dose dependent manner, which was stronger than that of arbutin used as a positive control.

Changes in the Isozyme Composition of Antioxidant Enzymes in Response to Aminotriazole in Leaves of Arabidopsis thaliana

Kyong-Suk Kang,Chang-Jin Lim,Tae-Jin Han,Joon-Chul Kim,Chang-Duck Jin 강원대학교 기초과학연구소 1999 기초과학연구 Vol.10 No.-

The changes in isozyme profiles of catalase (CAT), peroxidase (POD), ascorbate peroxidase (APX), and glutathione reductase(GR) during severe deactivation of total CAT activity by aminotriazole (AT) treatment were investigated in the leaves of Arabidopsis thaliana (Columbia ecolype) in relation to H₂O₂-mediated oxidative stress. In spite of striking deactivation of total CAT activity by 0.1 mM AT, there were no significant differences in H₂O₂levels or total leaf soluble protein contents including a Rubisco in both the control and AT-treated leaves. On the other hand, one specific protein band (molecular mass, 66 kD) was observed on the SDS-gel from leaf soluble proteins whose staining intensity was strikingly enhanced by AT treatment for 6 h. However, this band disappeared at 12 h. In the native-gel assays of CAT, POD, APX and GR isozymes, AT remarkably inhibited the expression of the CAT1 isozyme with no effects on CAT2 and CAT3, and generally had no effect on POD isozyme profiles. However, AT stmulated the intensity of activities of pre-existing APX1 and GR1 isozymes. In particular, it induced a new synthesis of one GR isozyme. Therefore, these results collectively suggest that a striking deactivation of total CAT activity by AT in A. thaliana leaves largely results from the suppression of CAT1 isozyme, and that APX1, GR1, and a newly syntheszed GR isozyme could complement the role of CAT1 to metabolize H₂O₂into non-toxic water.

α-Pinene Triggers Oxidative Stress and Related Signaling Pathways in A549 and HepG2 Cells

Kyong-Suk Jin,박민지,전미라,임호진,조완근,정우식 한국식품과학회 2010 Food Science and Biotechnology Vol.19 No.5

The volatile organic compounds (VOCs) −dlimonene,α-pinene, and isoprene− are widely used in scented products including food flavorings, air fresheners,and cosmetics. The effects of these VOCs on cell viability,oxidative stress generation, and the related molecular events were investigated in A549 and HepG2 cells. α-Pinene induced cell death, reactive oxygen species (ROS)generation, and reporter gene activities of antioxidant response element (ARE), activator protein 1 (AP-1), and nuclear factor (NF)-κB. α-Pinene stimulated the redoxsensitive transcription factors such as nuclear factor E2-related factor 2 (Nrf2) and NF-κB. Their down stream targets including heme oxygenase 1, inducible nitric oxide synthase, and cyclooxygenase 2 also increased after treatment with α-pinene. Moreover, α-pinene-induced oxidative stress-related signaling pathways could be connected by mitogen-activated protein kinases. Conversely,α-pinene-mediated cell death, ROS formation, and AP-1induction were inhibited by an antioxidant, N-acetyl-Lcysteine. Thus, α-pinene can induce the cell death possibly by modulating oxidative stress-related signaling pathways which can be reversed by an antioxidant treatment.

Molecular Mechanisms of 5-Azacytidine-Induced Trifluorothymidine-Resistance In Chinese Hamster V79 Cells

Jin, Kyong-Suk,Lee, Yong-Woo 대한의생명과학회 2005 Biomedical Science Letters Vol.11 No.2

A potent demethylating agent, 5-Azacytidine (5-AzaC) has been widely used as in many studier an DNA methylation, regulation of gene expression, and cancer biology. The mechanisms of the demethylating activity were known to be formation of complex between DNA and DNA methyltransferase (MTase), which depletes cellular MTase activity. However, 5-AzaC can also induce hypermethylation of a transgene in a transgenic cell line, G12 cells and it was explained as a result of defense mechanisms to inactivate foreign gene(s) somehow. This finding evoked the question that whether the phenomenon of hypermethylation induced by 5-AzaC is limited to the transgene or it can be occurred in endogenous gene(s). In order to answer the question, mutagenicity test of 5-AzaC and molecular characterization of mutants obtained from the test were performed using an endogenous gene, thymidine kinase (tk) in Chinese hamster V79 cells. When V79 and V79-J3 subclone cells were treated with 1, 2.5, 5, 10μM of 5-AzaC for 48 hours, their maximum mutant frequencies were revealed as 6×10^(-3) at 5μM (350-fold induction over background) and 8×10^(-3) at 2.5μM (1,800-fold induction over background) respectively. Since the induction rates were too high to be induced by true mutations, many trifluorothymidine (TFT)-resistant (TFT^(R)) cells were subjected to Northern blot analysis to check the presence of tk transcripts. Surprisingly, all clones tested possessed the transcripts in a similar level, that implicates the TFTR phenotype induced by 5-AzaC has not given rise to hypermethylation of the gene in spite of unusually high mutation frequency. In addition, it has shown that the TK activity in the Pool of 5-AzaC-induced TFT^(R) cells has about a half of that in spontaneously-induced TFT^(R) cells or in non-selected parental V79-J3 cells. This result suggests that the mechanism(s) underlying the TFT-resistance between spontaneously occurred and 5-AzaC-induced cells may be different. These findings have shown that the TFT^(R) phenotype induced by 5-AzaC has not given rise to hypermethylation of the tk gene, and 5-AzaC may be induced by one or combined pathways among many drug resistance mechanisms. The exact mechanisms for the 5-AzaC-induced TFT^(R) phenotype remain to elucidate.

Promises and Risks of Unsaturated Volatile Organic Compounds

Kyong-Suk Jin,Mira Jun,Min-Ji,Seon Ok,Jae-Han Jeong,Hye-Sook Kang,Wan-Keun Jo,Ho-Jin Lim,Woo-Sik Jeong 한국식품과학회 2008 Food Science and Biotechnology Vol.17 No.3

Limonene, pinene, and isoprene are abundant and ubiquitous volatile organic compounds (VOCs) which are found in various natural products and also produced from various manufacture processes. Limonene and pinene are major components of food additives and household products for enrichment of good flavors and elimination of malodors, and isoprene is a basal motif of monoterpenes such as limonene and pinene. They have shown many beneficial effects such as chemopreventive, chemotherapeutic, and antioxidant activities. Upon certain conditions, however, adverse effects of these compounds on human health have also been reported. Although they do not seem to have acute and severe toxicity to human, they can easily generate secondary organic aerosols (SOAs) when they react with oxygen and/or ozone, which have shown certain toxic effects on experimental animal models as well as on humans. Numerous household and scented products containing limonene, pinene, and isoprene are widely used in these days. However, biological consequences upon exposure to these products are largely unknown. The aim of this review is to summarize and analyze the current understanding on the biological effects of VOCs, in particular limonene, pinene, and isoprene, as well as their SOAs.

포황 메탄올 추출물에 의한 인체 대장암 세포주 HT29의 G2/M Arrest 및 Apoptosis 유발

진수정 ( Jin Soo Jung ),윤승근 ( Seung Geun Yun ),오유나 ( You Na Oh ),이지영 ( Ji Young Lee ),박현진 ( Hyun Jin Park ),진경숙 ( Kyong Suk Jin ),권현주 ( Hyun Ju Kwon ),김병우 ( Byung Woo Kim ) 한국미생물생명공학회(구 한국산업미생물학회) 2013 한국미생물·생명공학회지 Vol.41 No.4

본 연구에서는 인체 대장암세포인 HT29를 사용하여 포황 메탄올 추출물(Methanol extract of Typha orientalis, METO)의 항암 활성 및 그 분자적 기전에 관하여 분석하였다. 먼저 METO가 다양한 암세포의 증식에 미치는 영향을 분석한 결과, 인체 대장암 세포, 폐암 세포, 간암 세포 등의 세포증식을 억제하였으며 그 중에서도 대장암 세포인 HT29에 대해 강한 세포 증식 억제 효과를 나타내었다. METO에 의한 세포 증식 억제 기전을 분석하기 위하여 Flow cytometry analysis를 수행한 결과, METO 농도의존적으로 HT29 세포의 G2/M기 세포분포가 증가하고 아울러 apoptosis 유발군인 SubG1기 세포분포가 증가하는 것을 확인할 수 있었다. METO에 의한 HT29 세포의 G2/M arrest는 Cdc2의 inactive form인 phospho-Cdc2의 증가에 의한 G2/M checkpoint 관련 단백질의 활성억제에 의한 것이라 사료된다. 이러한 phospho-Cdc2의 증가는 METO에 의해 발현이 증가된 Wee1 kinase와 발현이 감소된 Cdc25C phosphatase에 의해 야기된 것임을 확인하였다. 또한 METO에 의한 HT29의 apoptosis 유도에 관한 분자적 기전 분석을 위해 Western blot analysis를 수행한 결과, METO 농도가 증가할수록 종양 억제 유전자인 p53, death receptor인 FAS, Bcl-2 family 중 pro-apoptotic 단백질인 Bax 및 cytosolic cytochrome C의 발현이 증가되고, Caspase-3가 활성화되어 단편화된 Caspase-3의 증가가 관찰되었다. 또한 활성화된 Caspase-3의 기질 단백질인 PARP의 단편화가 일어나 apoptosis가 유도되는 것을 알 수 있었다. 이상의 결과들로부터 METO는 인체 대장암세포 HT29의 G2/M arrest 및 apoptosis 유도에 의한 항암활성을 보유함을 확인하였다. Typha orientalis, also known as bulrush or cattail, is a perennial herbaceous plant found in freshwater wetlands and has been widely used in constructed wetlands for wastewater treatment. Recent data has revealed that SH21B, a mixture composed of seven herbs including T. orientalis, exhibited an anti-adipogenic activity by the inhibition of the expression of adipogenic regulators. However, the anti-cancer effect of T. orientalis and its molecular mechanisms remain unclear. In this study, we evaluated the anti-cancer effect and its mechanism in the methanol extract of T. orientalis (METO) on human colon carcinoma HT29 cells. It was found that METO treatment showed cytotoxic activity in a dose-dependent manner, and induced G2/M cell cycle arrest and apoptosis in HT29 cells. The induction of G2/M arrest by METO was associated with the up-regulation of phospho-Cdc2 (Tyr15), an inactive form of Cdc2 and the down-regulation of Cdc25c phosphatase. METO also induced tumor suppressor p53 and cyclin-dependent kinase inhibitor p21 (WAF1/CIP1) expression. In addition, METO-induced apoptosis was characterized by the proteolytic activation of caspase-3, degradation of poly ADP ribose polymerase (PARP), and up-regulation of death receptor FAS and pro-apoptotic Bax expression. Collectively, these results indicate that the cell cycle inhibition and apoptosis induction of METO in HT29 cells allows for the possibility of its use in anti-cancer therapies.

Euptelea pleiosperma 에탄올 추출물의 항산화 및 항염증 활성

진경숙 ( Kyong Suk Jin ),박정애 ( Jung Ae Park ),이지영 ( Ji Young Lee ),강지숙 ( Ji Sook Kang ),권현주 ( Hyun Ju Kwon ),김병우 ( Byung Woo Kim ) 한국미생물생명공학회(구 한국산업미생물학회) 2014 한국미생물·생명공학회지 Vol.42 No.2

본 연구에서는 Euptelea pleiosperma 에탄올 추출물 (EPEE)의 항산화능과 항염증 생리활성을 in vitro assay system 및 cell culture model system을 이용하여 분석하였다. 먼저 EPEE의 항산화능을 DPPH radical scavenging activity로 분석한 결과 radical 소거능의 정도가 양성 대조군으로 사용한 ascorbic acid 보다 높은 활성을 보여 매우 강한 항산화능을 보유함을 확인하였다. 또한 RAW 264.7 세포주를 이용한 H2O2 및 LPS의 유도에 의해 생성된 ROS에 대한 소거능을 분석한 결과에서도 농도의존적인 강한 소거능을 보였다. 뿐만 아니라 대표적인 항산화효소 중 하나로 천연물에 의한 항산화능 활성에 의해 주로 발현이 유도되는 hemeoxygenase 1 (HO-1) 및 그 전사 인자인 nuclear factor-E2-related factor 2 (Nrf2)의 단백질 발현이 EPEE의 처리에 의해 유의적으로 증가됨을 보였으며 이러한 HO-1 및 Nrf2의 발현 변화는 상위신호전달계인 MAPKs 및 PI3K/Akt에 의해 조절될 가능성을 보였다. 한편 EPEE가 LPS에 의해 유도된 NO 생성에 미치는 영향을 분석한 결과 농도의존적인 NO 생성 저해능을 보였으며 이는 NO 생성 단백질인 iNOS의 발현 저해에서 기인함을 확인하였다. 이와 같은 EPEE의 NO 생성 억제 효과는 염증 상위신호전달계인 NF- κB 및 AP-1의 조절을 통해 일어날 가능성을 보였다. 이러한 결과를 통해 EPEE의 높은 항산화능과 항염증 활성을 처음으로 확인하였으며 향후 기능성 소재로서 유용하게 활용될 수 있을 것으로 판단된다. In this study, the anti-oxidative and anti-inflammatory activities of Euptelea pleiosperma ethanol extract (EPEE) were evaluated using in vitro assays and cell culture model systems. EPEE possessed a more potent scavenging activity against 1,1-diphenyl- 2-picryl hydrazyl than the ascorbic acid used as a positive control. EPEE effectively suppressed lipopolysaccharide (LPS), in addition to hydrogen peroxide induced reactive oxygen species on RAW 264.7 cells. Furthermore, EPEE induced the expression of the anti-oxidative enzyme heme oxygenase 1 (HO-1) and its upstream transcription factor, nuclear factor-E2- related factor 2 (Nrf2), dose and time dependently. The modulation of HO-1 and Nrf2 expression might be regulated by mitogen-activated protein kinases and phosphatidyl inositol 3 kinase/Akt as their upstream signaling pathways. On the other hand, EPEE inhibited LPS induced nitric oxide (NO) formation without cytotoxicity. Suppression of NO formation was the result of the down regulation of inducible NO synthase (iNOS) by EPEE. Suppression of NO and iNOS by EPEE may be modulated by their upstream transcription factor, nuclear factor κB, and AP-1 pathways. Taken together, these results provide important new insights into E. pleiosperma, namely that it possesses anti-oxidative and anti-inflammatory activities, indicating that it could be utilized as a promising material in the field of nutraceuticals.

Endlicheria anomala (Nees) Mez 추출물의 항산화, 항염증 및 미백 활성

진경숙 ( Jin Kyong Suk ),이지영 ( Ji Young Lee ),권현주 ( Hyun Ju Kwon ),김병우 ( Byung Woo Kim ) 한국미생물생명공학회(구 한국산업미생물학회) 2013 한국미생물·생명공학회지 Vol.41 No.4

본 연구에서는 Endlicheria anomala (Nees) Mez 메탄올 추출물(EAME)의 항산화, 항염증 및 미백 생리활성을 in vitro assay 및 cell culture model system을 이용하여 분석 하였다. EAME의 항산화능을 분석한 결과 DPPH, H2O2로 유도한 ROS, LPS로 유도한 NO 등 다양한 산화적 스트레스원을 효과적으로 소거하였다. 대표적인 항산화 효소들로 천연물에 의한 항산화능 활성에 의해 주로 발현이 유도되는 세효소인 HO-1, TrxR1, NQO1 및 그 전사 인자인 Nrf2의 단백질 발현에 미치는 영향을 분석한 결과 시료 처리 농도의 증가에 따라 세 효소 및 Nrf2의 발현이 유의적으로 증가됨을 보였다. 또한 EAME는 in vitro DOPA oxidation을 강하게 저해하여 tyrosinase inhibitor로서 작용할 가능성을 시사 하였고 이에 B16F10 melanocyte를 이용하여 미백 효능을 분석한 결과 유의적인 melanin 생성억제능 및 tyrosinase 효소 활성 억제능을 보였으며 이는 tyrosinase, TRP-1, TRP-2 등 melanin 생성의 핵심 작용 효소들의 단백질 발현 저해를 통해 일어나는 것으로 나타났다. 이러한 결과를 통해 EAME가 높은 항산화능과 항염증 활성, 그리고 미백 활성을 보유함을 처음으로 밝혔으며 향후 기능성 식품 및 피부 미용 소재로서유용하게 활용될 수 있을 것으로 판단된다. In this study, the anti-oxidative, anti-inflammatory, anti-melanogenic activities of Endlicheria anomala (Nees) Mez methanol extract (EAME) were evaluated by use of in vitro assays and cell culture model systems. The results revealed that EAME scavenges various radicals such as 1,1-diphenyl-2-picryl hydrazyl hydrogen peroxide induced reactive oxygen species, and lipopolysaccharide induced nitric oxide. Furthermore, EAME induced the expression of anti-oxidative enzymes such as heme oxygenase 1, thioredoxin reductase 1, NAD(P)H dehydrogenase 1, and their upstream transcription factor, nuclear factor-E2- related factor 2. Moreover, EAME inhibited in vitro DOPA oxidation and 3-isobutyl-1-methylxanthine induced melanogenesis in B16F10 cells. Its anti-melanogenic activity will have originated from the inhibition of tyrosinase enzyme activity and melanogenesis related protein expression. Taken together, these results provide the important new insight that E. anomala possesses various biological activities such as anti-oxidative, anti-inflammatory, and anti-melanogenic. Therefore, it might be utilized as a promising material in the fields of nutraceuticals and cosmetics.

Decaisnea insignis 에탄올 추출물의 항산화 및 항염증 활성

진경숙(Kyong-Suk Jin),이지영(Ji Young Lee),권현주(Hyun Ju Kwon),김병우(Byung Woo Kim) 한국생명과학회 2014 생명과학회지 Vol.24 No.9

본 연구에서는 식물에 존재하는 신규 기능성 소재 개발의 일환으로 Decaisnea insignis 에탄올 추출물(DIEE)의 항산화 및 항염증 생리활성을 분석하였다. 먼저 DIEE의 항산화능을 DPPH 라디칼 소거능을 통해 분석한 결과 양성 대조군으로 사용한 아스코르빈산과 유사한 정도의 높은 소거활성을 보여 DIEE가 매우 강한 항산화능을 보유함을 확인하였다. 또한 RAW 264.7 세포주에서 H₂O₂ 및 LPS에 의해 유도된 ROS에 대한 DIEE의 소거능을 분석한 결과, 농도의존적인 강한 ROS 소거능을 보였다. 뿐만 아니라 대표적인 항산화효소 중 하나로 항산화능 보유천연물에 의해 발현이 유도되는 hemeoxygenase 1 (HO-1) 및 그 전사 인자인 nuclear factor-E2-related factor 2(Nrf2)의 단백질 발현이 DIEE의 처리에 의해 유의적으로 증가됨을 보였으며 이러한 HO-1 및 Nrf2의 발현 유도는 상위신호전달계인 MAPKs에 의해 조절될 가능성을 보였다. 한편 DIEE가 LPS에 의해 유도된 NO 생성에 미치는 영향을 분석한 결과 농도의존적인 NO 생성 저해능을 보였으며 이는 NO 생성 단백질인 iNOS의 발현 저해에서 기인함을 확인하였다. 이와 같은 DIEE의 NO 생성 억제 효과는 염증 상위신호전달계인 NF-κB 및 AP-1의 조절을 통해 일어날 가능성을 보였다. 이러한 결과를 통해 DIEE의 높은 항산화능과 항염증 활성을 처음으로 확인하였으며 향후 기능성 소재로서 유용하게 활용될 수 있을 것으로 판단된다. This study was conducted to explore new nutraceutical resources from the plant kingdom possessing biological activities. To fulfill this purpose, the anti-oxidative and anti-inflammatory activities of Decaisnea insignis ethanol extract (DIEE) were evaluated. First, DIEE possessed potent scavenging activity against 1,1-diphenyl-2-picryl hydrazyl (DPPH), similar to ascorbic acid used as a positive control. Moreover, DIEE inhibited lipopolysaccharide (LPS)- and hydrogen peroxide (H₂O₂)-induced reactive oxygen species (ROS) in RAW 264.7 cells. Furthermore, DIEE induced the expression of an anti-oxidative enzyme, heme oxygenase 1 (HO-1), and its upstream transcription factor, nuclear factor-E2-related factor 2 (Nrf2), in a dose-dependent manner. The modulation of the HO-1 and Nrf2 expressions might be regulated by mitogen-activated protein kinases (MAPKs) and their upstream signaling pathways. On the other hand, DIEE suppressed LPS-induced nitric oxide (NO) formation without cytotoxicity. The inhibition of the NO formation was the result of the down-regulation of inducible NO synthase (iNOS) by DIEE. The suppression of NO and iNOS by DIEE might be modulated by their upstream transcription factors, nuclear factor κB (NF-κB), and activator protein 1 (AP-1) pathways. Taken together, these results provide important new insights that D. insignis possesses anti-oxidative and anti-inflammatory activities. Therefore, it might be utilized as a promising material in the field of nutraceuticals.