• Title/Summary/Keyword: Extracellular enzymes

Search Result 339, Processing Time 0.026 seconds

Protective Effects of Bogol-tang on Monosodium Iodoacetate-induced Osteoarthritis and Interleukin-1β-treated Primary Chondrocytes (보골탕이 Monosodium Iodoacetate 유도 골관절염과 Interleukin-1β 유도 연골세포에 미치는 보호 효과)

  • Sung, Jin Wook;Lee, Hai Woong;Kang, Kyung Hwa;Kim, Kyoung Min;Cho, Sung Woo
    • Journal of Korean Medicine Rehabilitation
    • /
    • v.29 no.2
    • /
    • pp.101-113
    • /
    • 2019
  • Objectives Bogol-tang has clinically been used to protect joint cartilage and to treat osteoarthritis. Our objective was to study the protective effect of Bogol-tang extract (BGT) in functional impairment, behavioral disorders, cartilage loss and pathological changes in a monoiodoacetate (MIA)-induced murine osteoarthritis (OA) model and interleukin (IL)-$1{\beta}$ -treated primary rat chondrocytes. Methods Mouse knee joints were injected with MIA, a chemical that inhibits glycolysis and causes joint inflammation and matrix loss. MIA-OA induced mice orally administered BGT or acetaminophen (AAP) for 18 days by daily. Primary rat chondrocytes were pretreated with BGT or dexamethasone (DEX) and followed by co-incubation with IL-$1{\beta}$ (10 ng/mL). Results In MIA-OA mice model, BGT led to delayed response on hot plate analysis, and suppressed the cartilage loss and damages in joint tissues. BGT suppressed the elevated levels of inflammatory mediators, nitrite and $PGE_2$, the gene expression of matrix degrading enzymes, and extracellular-signal-regulated kinases 1/2 and c-JunN-terminal kinase phosphorylation in IL-$1{\beta}$-treated primary rat chondrocytes. Conclusions Our results suggest that BGT improve the knee joint function and delay the cartilage damages by anti-nociceptive, anti-inflammatory and ant-catabolic effects, which indicate BGT could be a potential candidate for osteoarthritis treatment.

Cultivable Microbial Diversity in Domestic Bentonites and Their Hydrolytic Enzyme Production

  • Seo, Dong-Ho;Cho, Eui-Sang;Hwang, Chi Young;Yoon, Deok Jun;Chun, Jeonghye;Jang, Yujin;Nam, Young-Do;Park, So-Lim;Lim, Seong-Il;Kim, Jae-Hwan;Seo, Myung-Ji
    • Microbiology and Biotechnology Letters
    • /
    • v.47 no.1
    • /
    • pp.125-131
    • /
    • 2019
  • We have isolated and identified 72 bacterial strains from four bentonite samples collected at the mining areas located in Gyeongsangbuk-do, Republic of Korea, and measured their hydrolytic enzyme (${\alpha}$-amylase, protease, and cellulase) activities to identify the isolates with industrial-use potential. Most of the isolates belonged to the Bacillaceae, with minor portions being from the Paenibacillaceae, Micrococcaceae, and Bacillales Family XII at the family level. Of the strains isolated, 33 had extracellular ${\alpha}$-amylase activity, 30 strains produced cellulase, and 35 strains produced protease. Strain MBLB1268, having the highest ${\alpha}$-amylase activity, was identified as Bacillus siamensis ($0.38{\pm}0.06U/ml$). Bacillus tequilensis MBLB1223, isolated from Byi33-b, showed the highest cellulase activity ($0.26{\pm} 0.04U/ml$), whereas Bacillus wiedmannii MBLB1197, isolated from Zdb130-b, exhibited the highest protease activity ($54.99{\pm}0.78U/ml$). These findings show that diverse bacteria of the Bacillaceae family adhere to and exist in bentonite and are potential sources of industrially useful hydrolytic enzymes.

Hepatoprotective Effect of Uncaria rhynchophylla on Thioacetamide-Induced Liver Fibrosis Model

  • Choi, Jeong Won;Shin, Mi-Rae;Lee, Ji Hye;Roh, Seong-Soo
    • Biomedical Science Letters
    • /
    • v.27 no.3
    • /
    • pp.142-153
    • /
    • 2021
  • Liver fibrosis is a wound-healing response to chronic liver injury, which is caused by the continuous and excess deposition of extracellular matrix (ECM). The aim of this study is to investigate whether Uncaria rhynchophylla water extract (UR) can ameliorate thioacetamide (TAA)-induced liver fibrosis. The liver fibrosis model was induced on C57BL/6 mice by intraperitoneal injection with TAA three times a week for 8 weeks. UR (200 mg/kg) or silymarin (50 mg/kg) was administered orally daily for 8 weeks. Biochemical analyses including AST, ALT, MPO, and Ammonia levels were measured in serum. In the mice liver tissues, western blot and histological staining were analyzed. As a result, UR dramatically reduced the levels in serum AST, ALT, MPO, and Ammonia levels. UR treatment regulated NADPH oxidase factors expression, and antioxidant enzymes except for GPx-1/2 were significantly increased via Nrf2 activation. Furthermore, pro-inflammatory mediators, such as COX-2 and iNOS were markedly suppressed through the inhibition of NF-κB activation. Expressions of ECM-related protein including α-SMA and Collagen I were noticeably decreased. The additional histological evaluation confirmed that hepatocyte damage and collagenous fiber accumulation were attenuated. Taken together, these data suggest that UR possessed hepatoprotective effects in TAA-induced liver fibrosis via the NF-κB inactivation and Nrf2 activation. Therefore, UR may act as a potential therapeutic drug against liver fibrosis.

Anti-inflammatory Effects of Actinidia Polygama Ethanol Extract in through the Regulated NF-κ B and MAPKs Activation in LPS Stimulated RAW 264.7 Cells (RAW 264.7 세포에 대한 NF-κ B와 MAPK 활성 억제를 통한 개다래 열매 에탄올 추출물의 항염증 효과)

  • Chung-Mu Park;Hyun-Seo Yoon
    • Journal of The Korean Society of Integrative Medicine
    • /
    • v.11 no.2
    • /
    • pp.119-128
    • /
    • 2023
  • Purpose : The fruit of Actinidia polygama has been used in oriental medicine for the treatment of gout, rheumatoid arthritis, and inflammation. Though A. polygama exhibited anti-inflammatory activity in RAW 264.7 cells and carrageenan-induced rat paw edema, the exact mechanism for anti-inflammation was not evaluated yet. In this study, the anti-inflammatory mechanisms of A. polygama ethanol extract (APEE) in lipopolysaccharide (LPS) stimulated RAW 264.7 cells. Methods : WST-1 assay was applied to analyze the cytotoxic effect of APEE in RAW 264.7 cells. The productions of nitric oxide (NO) and prostaglandin (PG) E2 were analyzed by the Griess reaction and enzyme immunoassay (EIA) assay, respectively. In addition, protein expressions for inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 were measured by Western blot analysis. The activated status of an inflammatory transcription factor, NF-κ B, and its upstream signaling molecules, mitogen-activated protein kinases (MAPKs), was also evaluated by Western blot analysis. Results : As a result, APEE treatment did not exhibit any cytotoxicity until the concentration of 200 ㎍/㎖. APEE treatment significantly inhibited NO and PGE2 productions as well as their enzymes, iNOS and COX-2 in a dose-dependent manner. The inflammatory transcription factor, NF-κ B, was also attenuated by APEE treatment. In addition, the phosphorylated status of MAPKs such as extracellular regulated kinase (ERK), c-jun NH2 kinase (JNK), and p38, were significantly diminished by APEE treatment in LPS stimulated RAW 264.7 cells. Conclusion : Consequently, APEE treatment significantly attenuated the production of inflammatory mediators and their enzyme expressions in LPS-stimulated RAW 264.7 cells. The inflammatory transcription factor, NF-κ B, and upstream signaling molecules, MAPKs, were also significantly attenuated by APEE treatment in LPS-activated RAW 264.7 cells. These results indicate that APEE might be a candidate to be utilized as a promising candidate for the treatment of inflammatory disorders.

α-Pinene Attenuates Methamphetamine-Induced Conditioned Place Preference in C57BL/6 Mice

  • Chan Lee;Jung-Hee Jang;Gyu Hwan Park
    • Biomolecules & Therapeutics
    • /
    • v.31 no.4
    • /
    • pp.411-416
    • /
    • 2023
  • Methamphetamine (METH) is a powerful neurotoxic psychostimulant affecting dopamine transporter (DAT) activity and leading to continuous excess extracellular dopamine levels. Despite recent advances in the knowledge on neurobiological mechanisms underlying METH abuse, there are few effective pharmacotherapies to prevent METH abuse leading to brain damage and neuropsychiatric deficits. α-Pinene (APN) is one of the major monoterpenes derived from pine essential oils and has diverse biological properties including anti-nociceptive, anti-anxiolytic, antioxidant, and anti-inflammatory actions. In the present study, we investigated the therapeutic potential of APN in a METH abuse mice model. METH (1 mg/kg/day, i.p.) was injected into C57BL/6 mice for four alternative days, and a conditioned place preference (CPP) test was performed. The METH-administered group exhibited increased sensitivity to place preference and significantly decreased levels of dopamine-related markers such as dopamine 2 receptor (D2R) and tyrosine hydroxylase in the striatum of the mice. Moreover, METH caused apoptotic cell death by induction of inflammation and oxidative stress. Conversely, APN treatment (3 and 10 mg/kg, i.p.) significantly reduced METH-mediated place preference and restored the levels of D2R and tyrosine hydroxylase in the striatum. APN increased the anti-apoptotic Bcl-2 to pro-apoptotic Bax ratio and decreased the expression of inflammatory protein Iba-1. METH-induced lipid peroxidation was effectively mitigated by APN by up-regulation of antioxidant enzymes such as manganese-superoxide dismutase and glutamylcysteine synthase via activation of nuclear factor-erythroid 2-related factor 2. These results suggest that APN may have protective potential and be considered as a promising therapeutic agent for METH-induced drug addiction and neuronal damage.

7α,25-Dihydroxycholesterol-Induced Oxiapoptophagic Chondrocyte Death via the Modulation of p53-Akt-mTOR Axis in Osteoarthritis Pathogenesis

  • Jeong-Yeon Seo;Tae-Hyeon Kim;Kyeong-Rok Kang;HyangI Lim;Moon-Chang Choi;Do Kyung Kim;Hong Sung Chun;Heung-Joong Kim;Sun-Kyoung Yu;Jae-Sung Kim
    • Molecules and Cells
    • /
    • v.46 no.4
    • /
    • pp.245-255
    • /
    • 2023
  • This study aimed to exploring the pathophysiological mechanism of 7α,25-dihydroxycholesterol (7α,25-DHC) in osteoarthritis (OA) pathogenesis. 7α,25-DHC accelerated the proteoglycan loss in ex vivo organ-cultured articular cartilage explant. It was mediated by the decreasing extracellular matrix major components, including aggrecan and type II collagen, and the increasing expression and activation of degenerative enzymes, including matrix metalloproteinase (MMP)-3 and -13, in chondrocytes cultured with 7α,25-DHC. Furthermore, 7α,25-DHC promoted caspase-dependent chondrocyte death via extrinsic and intrinsic pathways of apoptosis. Moreover, 7α,25-DHC upregulated the expression of inflammatory factors, including inducible nitric oxide synthase, cyclooxygenase-2, nitric oxide, and prostaglandin E2, via the production of reactive oxygen species via increase of oxidative stress in chondrocytes. In addition, 7α,25-DHC upregulated the expression of autophagy biomarkers, including beclin-1 and microtubule-associated protein 1A/1B-light chain 3 via the modulation of p53-Akt-mTOR axis in chondrocytes. The expression of CYP7B1, caspase-3, and beclin-1 was elevated in the degenerative articular cartilage of mouse knee joint with OA. Taken together, our findings suggest that 7α,25-DHC is a pathophysiological risk factor of OA pathogenesis that is mediated a chondrocyte death via oxiapoptophagy, which is a mixed mode of apoptosis, oxidative stress, and autophagy.

Arctigenin from Burdock Root Exhibits Potent Antibacterial and Anti-Virulence Properties against Pseudomonas aeruginosa

  • Abdulrahman E. Koshak;Mahmoud A. Elfaky;Hossam M. Abdallah;Dina A. I. Albadawi;Gamal A. Mohamed;Sabrin R. M. Ibrahim;Abdulrahim A. Alzain;El-Sayed Khafagy;Azza A. H. Rajab;Wael A. H. Hegazy
    • Journal of Microbiology and Biotechnology
    • /
    • v.34 no.8
    • /
    • pp.1642-1652
    • /
    • 2024
  • Arctium lappa (Burdock) root is used in various culinary applications especially in Asian Cuisine. Arctigenin (ARC) is a polyphenolic compound abundant in the roots of the burdock plant from which it derives its name. The emergence of bacterial resistance is a growing global worry, specifically due to the declining availability of new antibiotics. Screening for the antibacterial candidates among the safe natural products is a promising approach. The present study was aimed to assess the antibacterial activity of ARC against Pseudomonas aeruginosa exploring its effect on the bacterial cell membrane. Furthermore, the anti-virulence activities and anti-quorum sensing (QS) activities of ARC were in vitro, in vivo and in silico assessed against P. aeruginosa. The current results showed the ARC antibacterial activity was owed to its disruption effect of the cell membrane. ARC at sub-MIC significantly decreased the formation of biofilm, motility, production of extracellular enzymes and in vivo protected mice against P. aeruginosa. These anti-virulence activities of ARC are owed to its interference with bacterial QS and its expression. Furthermore, ARC showed mild effect on mammalian erythrocytes, low probability to induce resistance and synergistically combined with antibiotics. In summary, the promising anti-virulence properties of ARC indicate its potential as an effective supplement to conventional antibiotics for treating severe P. aeruginosa infections.

Isolation of Agarivorans sp. KC-1 and Characterization of Its Thermotolerant β-Agarase (한천분해세균 Agarivorans sp. KC-1의 분리 및 내열성 β-아가라제의 특성 규명)

  • Min, Kyung-Cheol;Lee, Chang-Eun;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
    • /
    • v.28 no.9
    • /
    • pp.1056-1061
    • /
    • 2018
  • This article reports an agar-degrading marine bacterium and characterizes its agarase. The agar-degrading marine bacterium, KC-1, was isolated from seawater on the shores of Sacheon, in Gyeongnam province, Korea, using Marine Broth 2216 agar medium. To identify the agar-degrading bacterium as Agarivorans sp. KC-1, phylogenetic analysis based on the 16S rRNA gene sequence was used. An extracellular agarase was prepared from a culture medium of Agarivorans sp. KC-1, and used for the characterization of enzyme. The relative activities at 20, 30, 40, 50, 60, and $70^{\circ}C$ were 65, 91, 96, 100, 77, and 35%, respectively. The relative activities at pH 5, 6, 7, and 8 were 93, 100, 87, and 82%, respectively. The extracellular agarase showed maximum activity (254 units/l) at pH 6.0 and $50^{\circ}C$ in 20 mM of Tris-HCl buffer. The agarase activity was maintained at 90% or more until 2 hr exposure at $20^{\circ}C$, $30^{\circ}C$ and $40^{\circ}C$, but it was found that the activity decreased sharply from $60^{\circ}C$. A zymogram analysis showed that Agarivorans sp. KC-1 produced 3 agar-degrading enzymes that had molecular weights of 130, 80, and 69 kDa. A thin layer chromatography analysis suggested that Agarivorans sp. KC-1 produced extracellular ${\beta}$-agarases as it hydrolyzed agarose to produce neoagarooligosaccharides, including neoagarohexaose (21.6%), neoagarotetraose (32.2%), and neoagarobiose (46.2%). These results suggest that Agarivorans sp. KC-1 and its thermotolerant ${\beta}$-agarase would be useful for the production of neoagarooligosaccharides that inhibit bacterial growth and delay starch degradation.

Production and Characterization of α-Galactosidases from Two Bacillus licheniformis Isolates (Bacillus licheniformis 분리균 2종의 α-Galactosidase 생산성과 효소특성)

  • Jin, Hyun Kyung;Yoon, Ki-Hong
    • Microbiology and Biotechnology Letters
    • /
    • v.43 no.3
    • /
    • pp.195-203
    • /
    • 2015
  • Two bacterial strains, Bacillus licheniformis YB-1413 and YB-1414, producing extracellular α-galactosidase, were obtained from homemade Doenjang. On the basis of their biochemical properties, 16S rRNA sequences and random amplified polymorphic DNA patterns by polymerase chain reaction, they were found to be somewhat different from one another. α-Galactosidase productivities of the two isolates were increased by wheat bran, but drastically decreased by melibiose, raffinose and sucrose which were used as carbon sources. The enzyme productivities were increased by yeast extract as a nitrogen source with maximum levels of 1.87 U/ml for YB-1413 and 1.69 U/ml for YB-1414, respectively. The enzymes of both isolates exhibited maximum activity for hydrolysis of para-nitrophenyl-α-D-galactopyranoside (pNP-αGal) under reaction conditions of pH 6.0 and 45℃. Their hydrolyzing activities for pNP-αGal were drastically decreased by the addition of low concentrations of ribose and galactose. They were capable of hydrolyzing completely α-1,6 linked galactosyl residue in melibiose, raffinose and stachyose, which are known to be anti-nutritional factors in products of soybean and legume. In relation to the latter, the isolates YB-1413 and YB-1414 have potential applicability in improving soybean-fermented foods and the nutritional value of soybean feed.

Effect of Tetracycline Analogues on The Activity of Matrix Metalloproteinase-3 in Gingival Fibroblasts (테트라사이클린계 약물이 치은섬유아세포내 MMP-3의 활성도에 미치는 영향)

  • Cho, Jong-Hee;Kim, Sang-Mok;Kim, Byung-Ock;Han, Kyung-Yoon
    • Journal of Periodontal and Implant Science
    • /
    • v.29 no.3
    • /
    • pp.677-693
    • /
    • 1999
  • Extracellular matrix component is degraded by enzymes of thematrix metalloproteinases(MMPs). MMPs are produced by both hemopoietic and structural cells. Increased activity of MMP-3 in periodontium is strongly associated with inflammatory periodontal disease. The purpose of the present study was to determine the effect of tetracycline analogues on the activity of MMP-3. Tetracycline-HCl, doxycycline-HCl, and minocycline-HCl were applied to huamn gingival fibroblasts at various concentrations of 10, 25, 50, 100, 200${\mu}g$/ml, and 1 hour later IL-$1{\beta}$ of 25ng/ml was added. After incubation for 24 hours the cells were reacted by enzyme-linked immunosorbent assay using proMMP-3 ELISA kit. The optical density was measured by microwell plate reader at 450nm. The relative activity of MMP-3 was calculated as the percentage of the optical density of each experimental group to that of the control. The difference of the optical density and the relative activity of MMP-3 between the experimental groups and the control wasstatistically analyzed by one way ANOVA. The results were as follows: 1. Tetracycline-HCl showed the tendency to inhibit the activity of MMP-3 at the concentration lower than 25${\mu}g$/ml, but increased significantly the activity of MMP-3 at the concentration of 200${\mu}g$/ml(p<0.05). 2. Doxycycline-HCl inhibited significantly the activity of MMP-3 at the concentration lower than 100${\mu}g$/ml, but increased significantly the activity of MMP-3 at the concentration of 200${\mu}g$/ml(p<0.05). 3. Minocycline-HCl inhibited the activity of MMP-3 at the concentration in the range of 10 to 200${\mu}g$/ml. Within the limit of the present study, the above results suggested that the low concentration of tetracycline analogues could inhibit the activity of MMP-3 induced by IL-$1{\beta}$ in human gingival fibroblasts.

  • PDF