• Title/Summary/Keyword: Enzyme inhibition

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Inhibition of endoplasmic reticulum stress in high-fat-diet-induced obese C57BL/6 mice: Efficacy of a novel extract from mulberry (Morus alba) leaves fermented with Cordyceps militaris

  • Lee, Mi Rim;Bae, Su Ji;Kim, Ji Eun;Song, Bo Ram;Choi, Jun Young;Park, Jin Ju;Park, Ji Won;Kang, Mi Ju;Choi, Hyeon Jun;Choi, Young Whan;Kim, Kyung Mi;Hwang, Dae Youn
    • Laboraroty Animal Research
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    • v.34 no.4
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    • pp.288-294
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    • 2018
  • A few clues about correlation between endoplasmic reticulum (ER) stress and mulberry (Morus alba) leaves were investigated in only the experimental autoimmune myocarditis and streptozotocin-induced diabetes. To investigate whether a novel extract of mulberry leaves fermented with Cordyceps militaris (EMfC) could suppress ER in fatty liver, alterations in the key parameters for ER stress response were measured in high fat diet (HFD)-induced obese C57L/6 mice treated with EMfC for 12 weeks. The area of adipocytes in the liver section were significantly decreased in the HFD+EMfC treated group as compared to the HFD+Vehicle treated group, while their level was higher in HFD+Vehicle treated group than No treated group. The level of the eukaryotic initiation factor 2 alpha ($eIF2{\alpha}$) and inositol-requiring enzyme 1 beta ($IRE1{\alpha}$) phosphorylation and CCAAT-enhancer-binding protein homologous protein (CHOP) expression were remarkably enhanced in the HFD+Vehicle treated group. However, their levels were restored in the HFD+EMfC treated group, although some differences were detected in the decrease rate. Similar recovery was observed on the ER stress-induced apoptosis. The level of Caspase-3, Bcl-2 and Bax were decreased in the HFD+EMfC and HFD+orlistat (OT) treated group compared to the HFD+Vehicle treated group. The results of the present study therefore provide first evidence that EMfC with the anti-obesity effects can be suppressed ER stress and ER stress-induced apoptosis in the hepatic steatosis of HFD-induced obesity model.

Triptolide Inhibits Lipopolysaccharide-Induced MUC5AC/5B Expression via Nuclear Factor-Kappa B in Human Airway Epithelial Cells (사람 호흡기 상피세포에서 Triptolide의 Nuclear Factor-Kappa B를 통한 Lipopolysaccharide로 유도된 MUC5AC/5B 발현 억제 효과)

  • Seo, Bo Hyeon;Choi, Tae Yeong;Choi, Yoon Seok;Bae, Chang Hoon;Na, Hyung Gyun;Song, Si-Youn;Kim, Yong-Dae
    • Korean Journal of Otorhinolaryngology-Head and Neck Surgery
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    • v.61 no.12
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    • pp.674-680
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    • 2018
  • Background and Objectives The representative mucin genes in the human airway are MUC5AC and MUC5B, which are regulated by several inflammatory and anti-inflammatory substances. Triptolide (TPL), udenafil, betulinic acid, changkil saponin, and glucosteroid are some of the many anti-inflammatory substances that exist. TPL is a diterpenoid compound from the thunder god vine, which is used in traditional Chinese medicine for treatment of immune inflammatory diseases, such as rheumatoid arthritis, systemic lupus erythematosus, nephritis and asthma. However, the effects of TPL on mucin expression of human airway epithelial cells have yet to be reported. Hence, this study investigated the effect of TPL on lipopolysaccharide (LPS)-induced MUC5AC and MUC5B expression in human airway epithelial cells. Subjects and Method The NCI-H292 cells and the primary cultures of human nasal epithelial cells were used to investigate the effects of TPL on LPS-induced MUC5AC and MUC5B expression using real-time polymerase chain reaction, enzyme immunoassay, and Western blot. Results TPL significantly decreased the LPS-induced MUC5AC and MUC5B mRNA expression and protein production. TPL also significantly decreased the nuclear factor-kappa B (NF-kB) phosphorylation. Conclusion These results suggest that TPL down regulates MUC5AC and MUC5B expression via inhibition of NF-kB activation in human airway epithelial cells. This study may provide important information about the biological role of triptolide on mucus-secretion in airway inflammatory diseases and the development of novel therapeutic agents for controlling such diseases.

Inhibitory Effects of Ethanol Extract of Rhodiola Sacra on Endoplasmic Reticulum Stress in Neuro-2A Cells (설치류 Neuro-2A 신경세포에서 홍경천 에탄올 추출물의 소포체 스트레스 억제효과)

  • Jo, Nam-Eun;Song, Young-soon
    • Journal of Digital Convergence
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    • v.17 no.8
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    • pp.265-270
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    • 2019
  • Growing evidence suggests that mediating apoptotic cell death of ER stress plays an important role in pathological development of neurodegenerative diseases including Alzheimer's disease. The ethanol extract of Rodiola sacra (ERS) investigates whether ER stress protects neuroinvasive neuro-2A cells from homocysteine (Hcy) cell death and ER stress. In neuronal cells, Hcy markedly decreased the viability of the cells and induced the death of Annexin V-positive cells as confirmed by MTT assay. The Hcy cell viability and apoptotic loss pretreated with ERS were attenuated, and Hcy showed stress in the expression of C / EBP homologous protein, 78-kDa glucose regulatory protein and the junction of X-box binding protein-1 (xbp1) mRNA. ESR decreased Hcy-induced mRNA binding, GRP78 and CHOP cells induced Hcy-induced ER stress and apoptosis, and Western blotting revealed expression of heme oxygenase-1 and HO-1 enzyme activity Inhibition is indicative of therapeutic value for neurodegenerative diseases such as decreased cell death by hemin.

The Effects of Heated Radish Extract on the Prevention of Ulcerative Colitis Inflammation (열처리된 무 추출물의 궤양성 대장염증 예방 효과에 미치는 영향)

  • Kim, Hyun Kyoung
    • The Journal of the Convergence on Culture Technology
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    • v.5 no.3
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    • pp.317-326
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    • 2019
  • The present study aimed to investigate the comparative evaluation of pharmacological efficacy between sulfasalazine alone and combination with herbal medicine on dextran sodium sulfate (DSS)-induced UC in mice. Balb/c mice received 5% DSS in drinking water for 7 days to induce colitis. Animals were divided into five groups (n = 9): group I-normal group, group II-DSS control group, group III-DSS + sulfasalazine (30 mg/kg), group IV-DSS + sulfasalazine (60 mg/kg), group V-DSS + sulfasalazine (30 mg/kg) + Radish Extract mixture (30 mg /kg) (SRE). DSS-treated mice developed symptoms similar to those of human UC, such as severe bloody diarrhea and weight loss. SRE supplementation, as well as sulfasalazine, suppressed colonic length and mucosal inflammatory infiltration. In addition, SRE treatment significantly reduced the expression of pro-inflammatory signaling molecules through suppression both MAPK) and nuclear factor-kappa B (NF-${\kappa}B$) signaling pathways, and prevented the apoptosis of colon. Moreover, SRE administration significantly led to the up-regulation of anti-oxidant enzyme including SOD and Catalase. This is the first report that Radish extract mixture combined with sulfasalazine protects against experimental UC via the inhibition of both inflammation and apoptosis, very similar to the standard-of-care sulfasalazin.

Systematic Target Screening Revealed That Tif302 Could Be an Off-Target of the Antifungal Terbinafine in Fission Yeast

  • Lee, Sol;Nam, Miyoung;Lee, Ah-Reum;Lee, Jaewoong;Woo, Jihye;Kang, Nam Sook;Balupuri, Anand;Lee, Minho;Kim, Seon-Young;Ro, Hyunju;Choi, Youn-Woong;Kim, Dong-Uk;Hoe, Kwang-Lae
    • Biomolecules & Therapeutics
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    • v.29 no.2
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    • pp.234-247
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    • 2021
  • We used a heterozygous gene deletion library of fission yeasts comprising all essential and non-essential genes for a microarray screening of target genes of the antifungal terbinafine, which inhibits ergosterol synthesis via the Erg1 enzyme. We identified 14 heterozygous strains corresponding to 10 non-essential [7 ribosomal-protein (RP) coding genes, spt7, spt20, and elp2] and 4 essential genes (tif302, rpl2501, rpl31, and erg1). Expectedly, their erg1 mRNA and protein levels had decreased compared to the control strain SP286. When we studied the action mechanism of the non-essential target genes using cognate haploid deletion strains, knockout of SAGA-subunit genes caused a down-regulation in erg1 transcription compared to the control strain ED668. However, knockout of RP genes conferred no susceptibility to ergosterol-targeting antifungals. Surprisingly, the RP genes participated in the erg1 transcription as components of repressor complexes as observed in a comparison analysis of the experimental ratio of erg1 mRNA. To understand the action mechanism of the interaction between the drug and the novel essential target genes, we performed isobologram assays with terbinafine and econazole (or cycloheximide). Terbinafine susceptibility of the tif302 heterozygous strain was attributed to both decreased erg1 mRNA levels and inhibition of translation. Moreover, Tif302 was required for efficacy of both terbinafine and cycloheximide. Based on a molecular modeling analysis, terbinafine could directly bind to Tif302 in yeasts, suggesting Tif302 as a potential off-target of terbinafine. In conclusion, this genome-wide screening system can be harnessed for the identification and characterization of target genes under any condition of interest.

Analysis on the Efficacy of Cosmetic Application of Lijang Snow Tea (Nekemias grossedentata) (리장 설차 (Nekemias grossedentata )의 화장품적 적용 효능 분석)

  • Wen, Ying;Lee, Seol-Hoon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.47 no.4
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    • pp.281-287
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    • 2021
  • In this study, we analyzed the cosmetic applicability of extract from snow tea, native to Lijiang, Yunnan-province, China. After confirming the species as N. grossedentata through DNA analysis of Lijiang snow tea, experiments were conducted using representative tea, green tea, and a representative control group for each efficacy analysis. Both teas were extracted using 70% (v/v) ethanol aqueous solution. The polyphenol content in the Lijiang snow tea extract (gallic acid equivalent, 23.9 ± 3.2 mg/mL) was higher than that in green tea extract (16.4 ± 2.3 mg/mL). In contrast, the antioxidant (Radical scavenging, IC50 104 ㎍/mL), tyrosinase enzyme inhibitory (whitening agent, IC50 40.7 ㎍/mL), and Escherichia coli growth inhibitory (preservative) activities (IC50 2.85 mg/mL) were analyzed based on the solid content in the extract, and it was confirmed that the activities of Lijiang snow tea extract were superior to those of green tea extract (radical scavenging, IC50 234 ㎍/mL. It also showed similar efficacy to previously used active substances such as antioxidants (vitamin C, IC50 108 ㎍/mL), whitening agents (vitamin C, IC50 80㎍/mL), and preservatives (methylparaben, IC50 4.35 mg/mL). However, green tea was found to be better in collagenase inhibition activity (anti-wrinkle). Through this study, the cosmetic application potential of Lijiang snow tea is high.

Anti-Graying Effect of Pueraria Lobata Root Extract on Stress-Induced Hair Graying (갈근 추출물의 스트레스성 백모 형성 억제 효과)

  • Hong, Min Jung;Park, Byung Cheol;Hong, Yong Deog;Kim, Su Na
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.48 no.3
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    • pp.287-293
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    • 2022
  • Gray hair is a representative sign of aging. Intrinsic aging, stress, and the external environment cause hair graying. Stress is known to be a major factor in the early onset of hair graying. We previously found that Pueraia lobata root extract (PLRE) can prevent hair graying by promoting melanin formation. However, it remains unknown whether PLRE can prevent hair graying induced by conditions of stress. In this study, we confirmed the effect of PLRE on stress-induced hair graying. A reporter cell line was newly constructed to confirm the expression of microphthalamia-associated transcription factor (MITF), the main transcription factor for melanin production. MITF expression and melanin pigmentation were reduced in human hair follicle tissue treated with the stress hormone cortisol or H2O2 to induce oxidative stress. PLRE treatment restored MITF expression and increased the amount of melanin pigment in the hair follicle. The expression of Tyrosinase related proteins-2 (TRP-2), a melanin synthesis enzyme in the hair follicle, also increased. In conclusion, PLRE can effectively prevent the inhibition of melanin synthesis by stress hormones and oxidative stress.

Anti-invasive Effect of Artemisia scoparia Halophyte Extract and its Solvent-partitioned Fractions in Human Fibrosarcoma Cells (인간 섬유육종세포에서 비쑥 추출물과 유기용매 분획물의 암전이 억제 효과)

  • Kim, Junse;Kong, Chang-Suk;Sim, Hyun-Bo;Seo, Youngwan
    • Journal of Life Science
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    • v.31 no.12
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    • pp.1100-1109
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    • 2021
  • The halophyte Artemisia scoparia is an edible medicinal plant, with insecticidal, anti-inflammatory, anticholesterol, antipyretic, and antibacterial effects. The aim of this study was to assess the inhibitory effect of crude extract and solvent-partitioned fractions obtained from A. scoparia on MMP-2 and MMP-9 activity in phorbol-12-myristate-13-acetate (PMA)-stimulated human fibrosarcoma HT-1080 cells using four different activity tests: gelatin zymography, MMP enzyme-linked immunosorbent assay (ELISA), wound healing assay, reverse transcription-polymerase chain reaction (RT-PCR), and Western blot assay. A. scoparia samples were extracted twice with methylene chloride (MC) and twice with methanol (MeOH). After the MC and MeOH crude extracts were combined, the combined crude extracts showed a significant inhibitory effect against MMP-2 and MMP-9 enzymes. They were then fractionated into n-hexane, 85% (v/v) aqueous methanol (85% (v/v) aq.MeOH), n-butanol, and water according to solvent polarity. Among the four solvent-partitioned fractions, n-hexane and 85% (v/v) aq. MeOH fractions significantly inhibited MMP-2 and MMP-9 activity and cell mobility. In addition, the n-hexane and 85% (v/v) aq.MeOH fractions effectively inhibited MMP-2 and -9 activity in the gelatin zymography and MMP ELISA assay. In the wound healing assay, RT-PCR, and Western blot assay, all solvent-partitioned fractions, except the H2O fraction, significantly suppressed cell migration, as well as the expression levels of MMP-2 and -9 mRNA and proteins.

Anti-inflammatory Effects of Ethanol Extract of Chinese Medicinal Plants in Yanjin on LPS-stimulated RAW 264.7 Macrophages (LPS로 자극한 RAW 264.7 세포에서 중국 연변에 자생하는 약용 식물 에탄올 추출물의 항염증 효과 연구)

  • Park, Yea-Jin;Seo, Jong-Hwan;Gil, Tae-Young;Cheon, Se-Yun;Piao, Ren-Zhe;Lee, Sang-Woo;Cha, Yun-Yeop;An, Hyo-Jin
    • The Korea Journal of Herbology
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    • v.33 no.6
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    • pp.71-78
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    • 2018
  • Objectives : This study was fulfilled to investigate nominee materials as anti-inflammatory agent from ethanol extract of Chinese medicinal plants in Yanjin. Among the 20 candidates, we selected most effective one, the ethanol extract of Cicuta virosa L. (CVL). The mechanism underlying the anti-inflammatory effects of CVL is not clearly identified as yet. Accordingly, we clarified the anti-inflammatory effects of CVL and its underlying molecular mechanisms in LPS-stimulated RAW 264.7 macrophages. Methods : RAW264.7 macrophages were incubated with CVL (12.5, 25, or $50{\mu}M$) and/or lipopolysaccharide (LPS) ($1{\mu}g/m{\ell}$). Cytotoxicity was determined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay and the level of nitric oxide (NO) production was measured with Griess reagent. The prostaglandin $E_2$ ($PGE_2$) production was measured with enzyme immunoassay kits and the protein expression of inducible nitric oxide synthase (iNOS) was determined using Western blot analysis. Results : Among the 20 ethanol extract of Chinese medicinal plants of Yanjin tested, CVL significantly reduced the production of NO in a dose-dependent manner via inhibition the protein expressions of iNOS without cytotoxicity on the LPS-stimulated RAW 264.7 macrophages. In addition, CVL also effectively declined the production of $PGE_2$ in LPS-simulated RAW 264.7 macrophages. Conclusions : Taken together, these data presented in this study demonstrate that CVL possesses anti-inflammatory activity by suppressing the production of pro-inflammatory mediators NO and $PGE_2$, and pro-inflammatory protein iNOS expression in LPS-stimulated RAW 264.7 macrophages.

Comparative Evaluation of Korean Medicine Well-matched with Pork through an Inhibition of Pancreatic Lipase (저육(猪肉)과 잘 어울리는 췌장라이페이즈 억제능을 가진 한약재의 비교평가)

  • Shin, Mi-Rae;Kim, SooHyun;Lee, AhReum;Kim, KyeongJo;Kim, SuJi;Lee, Young Cheol;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.32 no.5
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    • pp.57-63
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    • 2017
  • Objectives : The aim of this study is to search Korean Medicine with the superior inhibitory effect of pancreatic lipase including a possibility as a well-matched sauce material with pork. Methods : We chose 5 samples which have the potent inhibitory effect of pancreatic lipase enzyme among 9 samples. 5 samples were Glycyrrhizae rhizoma, Cinnamomi cortex, Ganoderma lucidum, Syzygium aromaticum, and Schisandra chinensis. Animals were divided into eight groups (n=7). The experimental groups except for normal group were fed 60% high-fat diet. 5 samples were orally administered at a dose of 200 mg/kg body weight and orlistat were orally administrated at a dose of 60 mg/kg body weight for 7 days. Biochemical anaylses of 5 samples were executed based on lipid parameters such as triglyceride (TG) and total cholesterol (TC). Results : Korean Medicines with $IC_{50}$ of below $1mg/m{\ell}$ were Glycyrrhizae rhizoma, Cinnamomi cortex, Ganoderma lucidum, Syzygium aromaticum, and Schisandra chinensis. Body weight change of all drug-treated groups except Glycyrrhizae rhizoma reduced significantly during the experimental period. Orlistat treatment reduced significantly both TG and TC in serum. The significant emission of TG in fece showed in Cinnamomi cortex treatment, whereas Ganoderma lucidum and Syzygium aromaticum showed a tendency to increase without a significance. Besides, Ganoderma lucidum and Schisandra chinensis emitted significantly TC to fece. Conclusions : In conclusion, Cinnamomi cortex may exert anti-obesity effect by directly inhibiting pancreatic lipase, which would prevent the absorption of lipid from the small intestine. Accordingly, Cinnamomi cortex may suggest a high possibility as a well-matched sauce material with pork.