• Title/Summary/Keyword: metabolites analysis

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CO/HO-1 Induces NQO-1 Expression via Nrf2 Activation

  • Kim, Hyo-Jeong;Zheng, Min;Kim, Seul-Ki;Cho, Jung-Jee;Shin, Chang-Ho;Joe, Yeon-Soo;Chung, Hun-Taeg
    • IMMUNE NETWORK
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    • v.11 no.6
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    • pp.376-382
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    • 2011
  • Background: Carbon monoxide (CO) is a cytoprotective and homeostatic molecule with important signaling capabilities in physiological and pathophysiological situations. CO protects cells/tissues from damage by free radicals or oxidative stress. NAD(P)H:quinone oxidoreductase (NQO1) is a highly inducible enzyme that is regulated by the Kelch-like ECH-associated protein 1 (Keap1)/nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE) pathway, which is central to efficient detoxification of reactive metabolites and reactive oxygen species (ROS). Methods: We generated NQO1 promoter construct. HepG2 cells were treated with CO Releasing Molecules-2 (CORM-2) or CO gas and the gene expressions were measured by RT-PCR, immunoblot, and luciferase assays. Results: CO induced expression of NQO1 in human hepatocarcinoma cell lines by activation of Nrf2. Exposure of HepG2 cells to CO resulted in significant induction of NQO1 in dose- and time-dependent manners. Analysis of the NQO1 promoter indicated that an antioxidant responsible element (ARE)-containing region was critical for the CO-induced Nrf2-dependent increase of NQO1 gene expression in HepG2 cells. Conclusion: Our results suggest that CO-induced Nrf2 increases the expression of NQO1 which is well known to detoxify reactive metabolites and ROS.

Effect of Drought Stress on the Concentration of Nitrogen Metabolites in White Clover

  • Kim, Tae-Hwan;Lee, Bok-Rye;Jung, Woo-Jin;Kim, Dae-Hyun;Kim, Kil-Yong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.47 no.2
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    • pp.95-101
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    • 2002
  • To investigate the changes in the composition and pool size of nitrogen metabolites under drought stress, white clover (Trifolium repens L.) were exposed to -0.04MPa (well-watered, control) or to -0.12MPa (drought-stressed) of soil water potential during 28 days. Dry weight of leaves in drought-stressed plants was remarkably decreased by 45% and 74% within 14 days and 28 days, respectively, compared with control. For nitrate concentration after 28 days of treatment, a significant difference (1.6 times higher in drought-stressed plants) was observed only in stolon. NH$_3$-NH$_4$$^{+}$ concentration in all three organs of drought-stressed plants linearly increased to more than 1.6 times higher level at 28 day when compared to the initial level (day 0), while the increasing rate in control was much less than that of drought-stressed plants. Proline concentrations in drought-stressed plants remarkably increased and reached to 7, 13 and 17 times higher level at 28 day compared to control. Protein concentration in leaves of drought-stressed plants tended to decrease, while it slightly increased during the first 14 days and reached a plateau afterward in control. There was not significant difference in the proteins concentration of stolon and roots throughout experimental period. On SDS-PAGE analysis, two major proteins specifically induced by drought stress (16-kD and 18-kD) were detected in stolon.n.

Metabolic classification of herb plants by NMR-based metabolomics

  • Kim, Hee-Eun;Choi, Ye Hun;Choi, Kwang-Ho;Park, Ji Su;Kim, Hyeon Su;Jeon, Jun Hyeok;Heu, Min Soo;Shin, Dong-Seon;Lee, Joon-Hwa
    • Journal of the Korean Magnetic Resonance Society
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    • v.16 no.2
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    • pp.91-102
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    • 2012
  • Metabolomics is the systematic identification and quantification of all metabolites in an organism or biological sample. NMR has been used as a major application tool in plant metabolomics such as quality control, chemotaxonomy, and analysis of genetically modified plants. Herbal medicines are the important therapeutics and are used to manage common diseases such as cold, inflammation, pain, heart diseases, liver cirrhosis, diabetes and central nerve system diseases. Herb plants include various kinds of species such as geranium, mint, and thyme and so on and contain different kinds of metabolites. We performed NMR-based metabolomics study on the seven different species of herb plants using $^1H$ NMR experiments and OPLS-DA to understand the correlation between the classification of herb plants and their metabolite contents. This study showed clear metabolic discrimination among various herb plants. This metabolmics study found several diagnostic NMR signals which are able to be used as bio-markers for identification of the specific herb plants among various species. Clear metabolic discrimination of herb plants suggests three chemotaxonomic groups of herb species.

Bioactive Compound Produced by Endophytic Fungi Isolated From Pelargonium sidoides Against Selected Bacteria of Clinical Importance

  • Manganyi, Madira Coutlyne;Tchatchouang, Christ-Donald K.;Regnier, Thierry;Bezuidenhout, Cornelius Carlos;Ateba, Collins Njie
    • Mycobiology
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    • v.47 no.3
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    • pp.335-339
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    • 2019
  • Endophytic fungi have the ability to live inside the host plant tissues without causing neither symptoms of diseases/or harm. Opportunistic infections are accountable for majority of the outbreaks, thereby putting a burden on the health system. To investigate and characterize the bioactive compounds for the control of bacteria of clinical importance, extracts from endophytic fungi were isolated from indigenous South African medicinal plants. Extracts from endophytic fungi were isolated from 133 fungal strains and screened against Gram positive and negative bacteria namely Bacillus cereus, Escherichia coli, Enterococcus faecium, and E. gallinarum using disk diffusion. Furthermore, gas chromatography-mass spectrometry was performed to identify the bioactive compounds. Sixteen out of one hundred and thirty-three (12%) fungi extracts exhibited antibacterial properties against some of the selected bacteria. E. coli was found to be the most susceptible in contrast to E. faecium and E. gallinarum which were the most resistant. The isolate MHE 68, identified as Alternaria sp. displayed the greater spectrum of antibacterial activities by controlling selected clinical bacteria strains including resistant E. faecium and E. gallinarum. The chemical analysis of the extract from MHE 68 indicated that linoleic acid (9,12-octadecadienoic acid (Z,Z)) and cyclodecasiloxane could be accountable for the antibacterial activity. This is the first study conducted on the secondary metabolites produced by endophytic fungal strains isolated from the Pelargonium sidoides DC. possessing antibacterial properties.

Biotransformation of Intestinal Bacterial Metabolites of Ginseng Saponin to Biologically Active Fatty-acid Conjugates

  • Hasegawa Hideo;Saiki Ikuo
    • Proceedings of the Ginseng society Conference
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    • 2002.10a
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    • pp.317-334
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    • 2002
  • Ginsenosides are metabolized (deglycosylated) by intestinal bacteria to active forms after oral administration. 20(S)-Protopanaxadiol $20-O-{\beta}-D-glucopyranoside$ (M1) and 20(S)-protopanaxatriol (M4) are the main intestinal bacterial metabolites (IBMs) of protopanaxadiol- and protopanaxatriol-type glycosides. M1 was selectively accumulated into the liver soon after its intravenous (i.v.) administration to mice, and mostly excreted as bile; however, some M1 was transformed to fatty acid ester (EMl) in the liver. EM1 was isolated from rats in a recovery dose of approximately $24mol\%.$ Structural analysis indicated that EM1 comprised a family of fatty acid mono-esters of M1. Because EM1 was not excreted as bile as Ml was, it was accumulated in the liver longer than M1. The in vitro cytotoxicity of M1 was attenuated by fatty acid esterification, implying that esterification is a detoxification reaction. However, esterified M1 (EM1) inhibited the growth of B16 melanoma more than Ml in vivo. The in vivo antitumor activity paralleled with the pharmacokinetic behavior. In the case of M4, orally administered M4 was absorbed from the small intestine into the mesenteric lymphatics followed by the rapid esterification of M4 with fatty acids and its spreading to other organs in the body and excretion as bile. The administration of M4 prior to tumor injection abrogated the enhanced lung metastasis in the mice pretreated with 2-chloroadenosine more effectively than in those pretreated with anti-asialo GMl. Both EM1 and EM4 did not directly affect tumor growth in vitro, whereas EM1 promoted tumor cell lysis by lymphocytes, particularly non-adherent splenocytes, and EM4 stimulated splenic NK cells to become cytotoxic to tumor cells. Thus, the esterification of IBM with fatty acids potentiated the antitumor activity of parental IBM through delay of the clearance and through immunostimulation. These results suggest that the fatty acid conjugates of IBMs may be the real active principles of ginsenosides in the body.

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Modified Renshen Wumei Decoction Alleviates Intestinal Barrier Destruction in Rats with Diarrhea

  • Guan, Zhiwei;Zhao, Qiong;Huang, Qinwan;Zhao, Zhonghe;Zhou, Hongyun;He, Yuanyuan;Li, Shanshan;Wan, Shifang
    • Journal of Microbiology and Biotechnology
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    • v.31 no.9
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    • pp.1295-1304
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    • 2021
  • Modified Renshen Wumei decoction (MRWD), a famous traditional Chinese medicine, is widely used for treating persistent diarrhea. However, as the mechanism by which MRWD regulates diarrhea remains unknown, we examined the protective effects of MRWD on intestinal barrier integrity in a diarrhea model. In total, 48 male rats were randomly distributed to four treatment groups: the blank group (CK group), model group (MC group), Medilac-Vita group (MV group) and Chinese herb group (MRWD group). After a 21-day experiment, serum and colon samples were assessed. The diarrhea index, pathological examination findings and change in ᴅ-lactate and diamine oxidase (DAO) contents illustrated that the induction of diarrhea caused intestinal injury, which was ameliorated by MV and MRWD infusion. Metabolomics analysis identified several metabolites in the serum. Some critical metabolites, such as phosphoric acid, taurine, cortisone, leukotriene B4 and calcitriol, were found to be significantly elevated by MRWD infusion. Importantly, these differences correlated with mineral absorption and metabolism and peroxisome proliferator-activated receptor (PPAR) pathways. Moreover, it significantly increased the expression levels of TLR4, MyD88 and p-NF-κB p65 proteins and the contents of IL-1 and TNF-α, while the expression levels of occludin, claudin-1 and ZO-1 proteins decreased. These deleterious effects were significantly alleviated by MV and MRWD infusion. Our findings indicate that MRWD infusion helps alleviate diarrhea, possibly by maintaining electrolyte homeostasis, improving the intestinal barrier integrity, and inhibiting the TLR4/NF-κB axis.

Study on Efficacy and Safety of High Dose MOK Pharmacopuncture in Hypothyroidism-induced Rats with Propylthiouracil (PTU 투여 갑상선기능저하증 유발 랫드모델에서 고용량 MOK 약침의 효능 및 안전성 평가 연구)

  • Hwang, Ji Hye;Im, Wu Hyun;Jung, Chul;Jung, Hyo Won
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.33 no.2
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    • pp.123-130
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    • 2019
  • In this study, we investigated the effects of MOK pharmacopuncture at high-doses which are increased 10 to 100-fold in clinics, on propylthiouracil (PTU)-induced hypothyroidism in rats and the safety. We measured the changes of body weight, food and water intake, body temperature, the serum levels of thyroid hormones (TSH, T3, and T4), AST and ALT, glucose, lipid metabolites (total cholesterol, HDL-cholesterol, LDL-cholesterol, and triglyceride) and observed histopathological changes of thyroid tissues by H&E staining. We also analyzed the peaks of constituents of MOK using HPLC. In the results, the treatment of MOK pharmacopuncture at high-dose (30 mg/kg) in hypothyroidism-induced rats for 2 weeks was shown the improvement effects on the decrease of body weight, food intake, and body temperature, The MOK pharmacopunture at high dose regulated the imbalance of thyroid hormones, glucose, and lipid metabolites and also inhibited the structural damages of thyroid tissues. In liver damage, the MOK pharmacopuncture at high dose reduced the increase of AST and ALT levels in hypothyroid rats. We identified the MOK constituents in HPLC analysis. In conclusion, the treatment of MOK pharmacopuncture at high dose has a therapeutic effect on hypothyroidism without liver toxicity, suggesting that the MOK pharmacopuncture be usefully applicable to treat with hypothyroidism in clinics.

Biotransformation of Reactive Red 141 by Paenibacillus terrigena KKW2-005 and Examination of Product Toxicity

  • Sompark, Chalermwoot;Singkhonrat, Jirada;Sakkayawong, Niramol
    • Journal of Microbiology and Biotechnology
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    • v.31 no.7
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    • pp.967-977
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    • 2021
  • A total of 37 bacterial isolates were obtained from dye-contaminated soil samples at a textile processing factory in Nakhon Ratchasima Province, Thailand, and the potential of the isolates to decolorize and biotransform azo dye Reactive Red 141 (RR141) was investigated. The most potent bacterium was identified as Paenibacillus terrigena KKW2-005, which showed the ability to decolorize 96.45% of RR141 (50 mg/l) within 20 h under static conditions at pH 8.0 and a broad temperature range of 30-40℃. The biotransformation products were analyzed by using UV-Vis spectrophotometry and Fourier-transform infrared spectroscopy. Gas chromatography-mass spectroscopy analysis revealed four metabolites generated from the reductive biodegradation, namely sodium 3-diazenylnaphthalene-1,5-disulfonate (I), sodium naphthalene-2-sufonate (II), 4-chloro-1,3,5-triazin-2-amine (III) and N1-(1,3,5-triazin-2-yl) benzene-1,4-diamine (IV). Decolorization intermediates reduced phytotoxicity as compared with the untreated dye. However, they had phytotoxicity when compared with control, probably due to naphthalene and triazine derivatives. Moreover, genotoxicity testing by high annealing temperature-random amplified polymorphic DNA technique exhibited different DNA polymorphism bands in seedlings exposed to the metabolites. They compared to the bands found in seedlings subjected to the untreated dye or distilled water. The data from this study provide evidence that the biodegradation of Reactive Red 141 by P. terrigena KKW2-005 was genotoxic to the DNA seedlings.

Acremonidin E produced by Penicillium sp. SNF123, a fungal endophyte of Panax ginseng, has antimelanogenic activities

  • Kim, Kyuri;Jeong, Hae-In;Yang, Inho;Nam, Sang-Jip;Lim, Kyung-Min
    • Journal of Ginseng Research
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    • v.45 no.1
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    • pp.98-107
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    • 2021
  • Background: Ginseng extracts and ginseng-fermented products are widely used as functional cosmetic ingredients for their whitening and antiwrinkle effects. Recently, increasing attention has been given to bioactive metabolites isolated from endophytic fungi. However, little is known about the bioactive metabolites of the fungi associated with Panax ginseng Meyer. Methods: An endophytic fungus, Penicillium sp. SNF123 was isolated from the root of P. ginseng, from which acremonidin E was purified. Acremonidin E was tested on melanin synthesis in the murine melanoma cell line B16F10, in the human melanoma cell line MNT-1, and in a pigmented 3D-human skin model, Melanoderm. Results: Acremonidin E reduced melanogenesis in α-melanocyte-stimulating hormone (α-MSH)-stimulated B16F10 cells with minimal cytotoxicity. qRT-PCR analysis demonstrated that acremonidin E downregulated melanogenic genes, including tyrosinase and tyrosinase-related protein 1 (TRP-1), while their enzymatic activities were unaffected. The antimelanogenic effects of acremonidin E were further confirmed in MNT-1 and a pigmented 3D human epidermal skin model, Melanoderm. Immunohistological examination of the Melanoderm further confirmed the regression of both melanin synthesis and melanocyte activation in the treated tissue. Conclusion: This study demonstrates that acremonidin E, a bioactive metabolite derived from a fungal endophyte of P. ginseng, can inhibit melanin synthesis by downregulating tyrosinase, illuminating the potential utility of microorganisms associated with P. ginseng for cosmetic ingredients.

A comparative analysis of rumen pH, milk production characteristics, and blood metabolites of Holstein cattle fed different forage levels for the establishment of objective indicators of the animal welfare certification standard

  • Baek, Dong Jin;Kwon, Hyoun Chul;Mun, Ah Lyum;Lim, Joo Ri;Park, Sung Won;Han, Jin Soo
    • Animal Bioscience
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    • v.35 no.1
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    • pp.147-152
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    • 2022
  • Objective: This study was conducted to obtain an objective index that can be quantified and used for establishing an animal welfare certification standard in Korea. For this purpose rumen pH, ruminating time, milk yield, milk quality, and blood components of cows reared in farms feeding high forage level (90%) and farms feeding low forage level (40%) were compared. Methods: Data on rumen pH, rumination time, milk yield, milk fat ratio, milk protein ratio, and blood metabolism were collected from 12 heads from a welfare farm (forage rate 88.5%) and 13 heads from a conventional farm (forage rate 34.5%) for three days in October 2019. Results: The rumination time was longer in cattle on the welfare farm than on the conventional farm (p<0.01), but ruminal pH fluctuation was greater in the cattle on conventional farm than the welfare farm (p<0.01). Conventional farms with a high ratio of concentrated feed were higher in average daily milk yield than welfare farms, but milk fat and milk production efficiency (milk fat and milk protein corrected milk/total digestible nutrients) was higher in cattle on welfare farms. Blood test results showed a normal range for both farm types, but concentrations of total cholesterol and non-esterified fatty acid were significantly higher in cows from conventional farms with a high milk yield (p<0.01). Conclusion: The results of this study confirmed that cows on the animal welfare farm with a high percentage of grass feed had higher milk production efficiency with healthier rumen pH and blood metabolism parameters compared to those on the conventional farm.