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Effects of Jinmu-tang on the Osteoarthritis by MIA in Rats (진무탕(眞武湯)이 MIA 유도 골관절염 흰쥐 모델에 미치는 영향)

  • Yang, Doo-Hwa;Woo, Chang-Hoon;An, Hee-Duk
    • Journal of Korean Medicine Rehabilitation
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    • v.28 no.1
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    • pp.19-31
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    • 2018
  • Objectives The object of this study was to investigate the antioxidative and antiinflammatory effects of Jinmu-tang extract (JMT) on the Monosodium iodoacetate (MIA)-induced rat osteoarthritis. Methods To investigate the antioxidant capacities of JMT, we measured the total polyphenol and flavonoid, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-Azino-bis(3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity. To evaluate the antioxidative and antiinflammatory effects of JMT, the rats were divided into 5 groups (n=8). Normal group was not induced by MIA and treated at all (N), control group was induced by MIA and not treated at all (Con), positive control group was induced by MIA and orally administered indomethacin 5 mg/kg (Indo) and experimental groups were induced by MIA and orally administered JMT 100 mg/kg (JMT100) and JMT 200 mg/kg (JMT200) for 4 weeks. The changes of anti-type II collagen antibody in serum, heme oxygenase-1 (HO-1), phosphorylated inhibitor of ${\kappa}B{\alpha}$ ($p-I{\kappa}B{\alpha}$), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS) and tumor necrosis factor alpha ($TNF-{\alpha}$) in knee joint tissue and histopathological observation (Hematoxylin & Eosin and Safranin-O stain) were measured. Results Total polyphenol and flavonoid levels of JMT were $26.90{\pm}0.33mg/g$ and $6.02{\pm}0.34mg/g$. $IC_{50}$ of L-ascorbic acid and JMT of DPPH radical scavenging activity were $1.35{\pm}0.07{\mu}g/ml$ and $52.95{\pm}0.97{\mu}g/ml$. $IC_{50}$ of L-ascorbic acid and JMT of ABTS radical scavenging activity were $3.18{\pm}0.02{\mu}g/ml$ and $91.49{\pm}1.74{\mu}g/ml$. In serum, the anti-type II collagen antibody levels of JMT100 and JMT200 groups were decreased significantly. In knee joint tissue, the HO-1 level of JMT200 was increased significantly. The $p-I{\kappa}B{\alpha}$ and $TNF-{\alpha}$ levels of JMT200 were decreased significantly. The COX-2 and iNOS levels of JMT groups were decreased significantly. In histopathological observation, in comparison with Con, synovial tissue, cartilage and proteoglycan of JMT100 and JMT200 were well preserved. Conclusions According to the results, It is considered that JMT has antioxidant and antiinflammatory effects for MIA-induced rat osteoarthritis, so it could be applied to osteoarthritis treatment.

Effects of Woohwangcheongsim-won on Gene Expression in a Hypoxic Model of Cultured Rat Cortical Cells (배양한 흰쥐 대뇌세포의 저산소증 모델에서 우황청심원이 유전자 표현에 미치는 영향)

  • Park Dong-Wan;Kim Wan-Sik;Bae Cheol-hwan;Jeong Sung-Hyun;Shin Gil-Cho;Lee Won-Chul
    • The Journal of Korean Medicine
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    • v.25 no.3
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    • pp.123-136
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    • 2004
  • Objectives : The purpose of this investigation is to evaluate the effects of Woohwangcheongsim-won (WC) on the in vitro neuronal development and alteration in gene expression in a hypoxia model using cultured rat cortical cells. Methods : E/sub 18/ rat cortical cells were grown in a neurobasal medium containing B27 supplement and various concentration of WC. Initial development of growth cone was investigated by phase-contrast microscopy, while dendritic spine formation and synaptogenesis were investigated by immunocytochemistry with SynGAPα(a postsynaptic marker) and synaptophysin (presynaptic marker) antibodies. Alteration in gene expression was analyses by microarray using rat 5K-TwinChips. Results : WC suppressed the development of growth cones and WC increased the number of dendritic spines at 20 and 50㎍/mL concentration but there was no statistical significance. Instead, it significantly decreased the number at 100㎍/mL. The expression of anti-apoptosis gene Bcl2-like 1 (Bcl211) increased (Global M=0.46), while Akt1 decreased. Proapoptosis genes Bad and PDCD2 increased. The expression of hemoglobin alpha 1 (probably neuroglobin) increased (Global M=0.93). The expression of antioxidants such as catalase, heme oxygenase (HO), and PRKAG2 gene increased. The expression PKC gene increased. The expression of retinoic acid receptor alpha (RARα) increased significantly (Global M=1.0). Conclusions : These data suggest that WC trends to suppress cellular activity slightly in normoxia and increases the expression of apoptosis-, antioxidation-, oxygen capture-related genes in hypoxia, but increases Bcl111 that anti-apoptosis gene, on the other hand increases Bad, PDCD2 that pro-apoptosis genes, too..

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Cloning and Characterization of Monofunctional Catalase from Photosynthetic Bacterium Rhodospirillum rubrum S1

  • Lee, Dong-Heon;Oh, Duck-Chul;Oh, You-Sung;Malinverni, Juliana C.;Kukor, Jerome J.;Kahng, Hyung-Yeel
    • Journal of Microbiology and Biotechnology
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    • v.17 no.9
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    • pp.1460-1468
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    • 2007
  • In this study, an approx. 2.5-kb gene fragment including the catalase gene from Rhodospirillum rubrum S1 was cloned and characterized. The determination of the complete nucleotide sequence revealed that the cloned DNA fragment was organized into three open reading frames, designated as ORF1, catalase, and ORF3 in that order. The catalase gene consisted of 1,455 nucleotides and 484 amino acids, including the initiation and stop codons, and was located 326 bp upstream in the opposite direction of ORF1. The catalase was overproduced in Escherichia coli UM255, a catalase-deficient mutant, and then purified for the biochemical characterization of the enzyme. The purified catalase had an estimated molecular mass of 189 kDa, consisting of four identical subunits of 61 kDa. The enzyme exhibited activity over a broad pH range from pH 5.0 to pH 11.0 and temperature range from $20^{\circ}C$ to $60^{\circ}C$C. The catalase activity was inhibited by 3-amino-1,2,4-triazole, cyanide, azide, and hydroxylamine. The enzyme's $K_m$ value and $V_{max}$ of the catalase for $H_2O_2$ were 21.8 mM and 39,960 U/mg, respectively. Spectrophotometric analysis revealed that the ratio of $A_{406}$ to $A_{280}$ for the catalase was 0.97, indicating the presence of a ferric component. The absorption spectrum of catalase-4 exhibited a Soret band at 406 nm, which is typical of a heme-containing catalase. Treatment of the enzyme with dithionite did not alter the spectral shape and revealed no peroxidase activity. The combined results of the gene sequence and biochemical characterization proved that the catalase cloned from strain S1 in this study was a typical monofunctional catalase, which differed from the other types of catalases found in strain S1.

Effects of Scutellaria baicalensis GEORGI on Gene Expression in a Hypoxic Model of Cultured Rat Cortical Cells (배양한 흰쥐 대뇌세포의 저산소증 모델에서 황금(黃芩)이 유전자 표현에 미치는 영향)

  • Chung, Sung-Hyun;Shin, Gil-Cho;Lee, Won-Chul;Kim, Sung-Bae
    • The Journal of Internal Korean Medicine
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    • v.25 no.4
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    • pp.324-336
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    • 2004
  • Objectives : The purpose of this investigation is to evaluate the effects of Scutellaria baicalensis GEORGI on alteration in gene expression in a hypoxia model using cultured rat cortical cells. Methods : E18 rat cortical cells were grown in a Neurobasal medium containing B27 supplement. On 12 DIV, Scutellaria baicalensis GEORGI(20 ug/ml) was added to the culture media and left for 24 hrs. On 11 DIV, cells were given a hypoxic insult $(2%\;O_2/5%\;CO_2,\;37^{\circ}C,\;3\;hrs)$, returned to normoxia and cultured for another 24 hrs. Total RNA was prepared from Scutellaria baicalensis GEORGI-untreated (control) and -treated cultures and alteration in gene expression was analysed by microarray using rat 5K-TwinChips. Results : For most of the genes altered in expression, the Global M values were between -0.5 to +0.5. Among these, 1143 genes increased in their expression by more than Global M +0.1, while 1161 genes decreased by more than Global M -0.1. Effects on some of the genes whose functions are implicated in neural viability are as follows: 1) The expression of apoptosis-related genes such as Bad (Global M = 0.39), programmed cell death-2(Pdcd2) (Global M = 0.20) increased, while Purinergic receptor P2X(P2rxl) Global M = -0.22), Bc12-like1(Bc1211)(Global M = -0.19) decreased. 2) The expression of 'response to stress-related genes such as antioxidation-related AMP-activated protein kinase subunit gamma 1 gene (Prkag1) (Global M = 0.14), catalase gene (Global M = 0.14) and Heme Oxygenase(Hmoxl) increased. 3) The expression of Fos like antigen 2 (Fos12) expressed in neurons that survive ischemic insult increased (Global M = 0.97). Conclusions : these data suggest that Scutellaria baicalensis GEORGI increases the expression of antiapoptosis- and antioxidation- related genes in a way that can not yet be explained.

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Ethanol Extract from Asparagus Cochinchinensis Attenuates Glutamate-Induced Oxidative Toxicity in HT22 Hippocampal Cells (HT22 해마세포의 oxidative toxicity에 대한 천문동 유래 에탄올추출물의 보호 효과)

  • Pak, Malk Eun;Choi, Byung Tae
    • Journal of Life Science
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    • v.26 no.12
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    • pp.1458-1465
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    • 2016
  • We investigated the neuroprotective effect of an ethanol extract from Asparagus cochinchinensis (AC) against glutamate-induced toxicity in the HT22 hippocampal cell, which is an ideal in vitro model for oxidative stress. The neuroprotective effects of AC in HT22 cells were evaluated by analyzing cell viability, lactate dehydrogenase (LDH), flow cytometry for cell death types, reactive oxygen species (ROS), mitochondria membrane potential (MMP), and Western blot assays. In the cell death analysis, AC treatment resulted in significantly attenuated glutamate-induced loss of cell viability with a decrease in LDH release. AC treatment also reduced glutamate-induced apoptotic cell death. In the ROS and MMP analysis, AC treatment inhibited the elevation of intracellular ROS induced by glutamate exposure and the disruption of MMP. In oxidative stress-related proteins analysis, AC treatment inhibited the expression of poly ADP ribose polymerase and heme oxygenase-1 by glutamate. These results indicate that AC exerts a significant neuroprotective effect against glutamate-induced hippocampal damage by decreasing ROS production and stabilizing MMP. Thus, AC potentially provides a new strategy for the treatment of oxidative stress-related diseases.

Cloning and Characterization of Soybean IFS (Isoflavone Synthase) Genes from Korean Cultivar, Sinpaldalkong (신팔달콩 유래 IFS (isoflavone synthase)유전자 클로닝 및 기능 규명)

  • Park, Hayng-Mi;Shin, Sang-Hyun;Ko, Jong-Min;Yi, Gi-Hwan;Nam, Min-Hee;Chung, Young-Soo;Chung, Won-Bok;Lee, Jai-Heon;Park, Seong-Whan
    • Journal of Life Science
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    • v.14 no.1
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    • pp.38-44
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    • 2004
  • Two genes, SinIFS1 and SinIFS2 from Korean soybean cultivar, Sinpaldalkong known as one of isoflavonerich cultivars, were cloned with PCR and degenerate primers. The sequences of two genes were analyzed with previously reported IFS genes of leguminous plants and their expression pattern in various environmental conditions was surveyed. The genomic clone of SinIFS1 contained 1,828bp nucleotides and encoded a polypeptide of 521 amino acids, and 1912bp nucleotides and a polypeptide of 521 amino acids for SinIFS2. Both genes included several conserved motifs, oxygen binding and activation (A/G-G-X-E/D-T-T/S), ERR triad (E...R....R), and heme binding (F-X-X-G-X-R-X-C-X-G) domain, which are typical in any member of cytochrome P45O superfamily. Very high sequence homology (>98%) was observed in the comparison with other IFSs of legumes. In the northern blot analysis to check the expression and increase of SinIFS1 to various environmental renditions (low temperature, light, dark, UV, and fungal elicitor), the most significant induction, more than 6 times of transcript level compared to the dark treatment as a control, was observed from the fungal elicitor treatment. The next up-regulated expression was from UV treatment (4${\times}$), low temperature and light conditions.

Nutritional Status of Iron of Elderly in Jeon-Ju Area (전주지역 노인의 철분영양상태)

  • 주은정;김인숙;서은아
    • Korean Journal of Community Nutrition
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    • v.5 no.3
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    • pp.493-501
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    • 2000
  • The purpose of this study was to assess the nutritional status of iron of the elderly residing in the Jeonju area. The anthropometric parameters, nutrient intake and biochemical status of iron, were measured for 60 korean elderly(23 elderly men and 37 elderly women aged 60 - 79 years old). The level of hemoglobin(Hb), hematocrint(Hct), serum iron(Fe), total iron binding capacity(TIBC) and serum ferritin(Ferritin) were measured and transferrin saturation(TFsaturation) was calculated. Mean values of Hb, Hct, Fe, TIBC, TFsaturation and Ferritin were 14.49$\pm$0.93g/이, 42.47$\pm$2.59%, 125.48$\pm$52,.46$\mu\textrm{g}$/dl, 338.13$\pm$45.92$\mu\textrm{g}$/dl, 193.00$\pm$125.03$\mu\textrm{g}$/l in elderly men and 12.82$\pm$0.99$\mu\textrm{g}$/dl, 37.66$\pm$2.90%, 100.08$\pm$42.82$\mu\textrm{g}$/dl, 356.41$\pm$54.65$\mu\textrm{g}$/dl, and 99.35$\pm$117.22$\mu\textrm{g}$/l in elderly women, respectively. Prevalence of iron deficiency varied greatly with biochemical indices of iron. It was 13.0% when judged by Fe(60 $\mu\textrm{g}$/dl) and TFsaturation(15%) whereas 34.78% by Hct(41%) in elderly men. However 13.5% of the elderly women showed iron depletion(Ferritin<20$\mu\textrm{g}$/l) and 43.2% showed suppressed erythropoiesis with iron deficiency(TIBC>360$\mu\textrm{g}$/dl). The anemic subjects assessed with TFsaturation(<15%) represented 13.5% of the elderly women, whereas 18.9% of the subjects possessed less than 12g/dl of Hb. The Hb concentration was positively correlated with Hct(r=0.980, p<0.001), Fe(r=0.384, p<0.01) and TFsaturation(r=0.349, p<0.01). On the other hand, Ferritin concentration showed a significantly negative correlation with TIBC(r=0.349, p<0.05) and a positive correlation with TFsaturation(r=0.362, p<0.01). Major food groups of iron intake in the elderly were vegetables, cereals, and fish. The mean daily intake of iron was not significantly different between elderly men and women(12.82mg vs 10.35mg). Intake of heme iron however, was significantly higher(p<0.01) in elderly men(1.03mg) than women(0.42mg). Total absorbable iron caculated by the method of Monsen was 0.55mg, 0.40mg in elderly men and women, respectively and bioavailability of dietary iron 4.29% and 3.87%.

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Antioxidative and Anti-inflammatory Activities of Ardisia arborescens Ethanol Extract (Ardisia arborescens 에탄올 추출물의 항산화 및 항염증 활성)

  • Jin, Kyong-Suk;Lee, Ji Young;Kwon, Hyun Ju;Kim, Byung Woo
    • Journal of Life Science
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    • v.24 no.7
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    • pp.713-720
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    • 2014
  • In this study, the antioxidative and anti-inflammatory activities of Ardisia arborescens ethanol extract (AAEE) were evaluated using in vitro assays and a cell culture model system. AAEE exhibited potent scavenging activity against 1,1-diphenyl-2-picryl hydrazyl (DPPH), similar to ascorbic acid, which was used as a positive control. Moreover, AAEE effectively suppressed lipopolysaccharide (LPS)- and hydrogen peroxide ($H_2O_2$)-induced reactive oxygen species (ROS) in RAW 264.7 cells. Furthermore, AAEE induced the expression of antioxidative enzymes, heme oxygenase 1 (HO-1), and thioredoxin reductase 1 (TrxR1), in addition to their upstream transcription factor, nuclear factor-E2-related factor 2 (Nrf2), in a dose-dependent manner. The upstream signaling pathways of mitogen-activated protein kinases (MAPKs) might regulate the modulation of HO-1, TrxR1, and Nrf2 expression. On the other hand, AAEE inhibited LPS-induced nitric oxide (NO) formation, without cytotoxicity. Suppression of NO formation was the result of AEEE-induced down-regulation of inducible NO synthase (iNOS). The suppression of NO and iNOS by AAEE might be modulated by their upstream transcription factor, nuclear factor (NF)-${\kappa}B$, and activator protein (AP)-1 pathways. Taken together, these results provide important new insights into the antioxidative and anti-inflammatory activities of A. arborescens. AAAEE might represent a promising material in the field of nutraceuticals.

The Anti-oxidative and Anti-inflammatory Activities of Malus melliana Ethanol Extract (Malus melliana 에탄올 추출물의 항산화 및 항염증 활성)

  • Lee, Su Hyeon;Jin, Kyong-Suk;Kim, Byung Woo;Kwon, Hyun Ju
    • Journal of Life Science
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    • v.27 no.7
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    • pp.783-789
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    • 2017
  • Malus melliana (Hand.-Mazz.) Rehder (M. melliana) is a Chinese plant that belongs to the Rosaceae family. There have been no previous reports regarding its bioactivity. In this study, the anti-oxidative and anti-inflammatory activities of M. melliana ethanol extract (MMEE) were evaluated using a 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging activity assay, reactive oxygen species (ROS) scavenging activity assay, nitric oxide (NO) inhibitory activity assay, and the analysis of related protein expressions through Western blot hybridization. MMEE showed potent scavenging activity against DPPH, similar to ascorbic acid, a well-known anti-oxidative agent, which was used as a positive control. MMEE also inhibited hydrogen peroxide-induced ROS in RAW 264.7 cells. Moreover, MMEE induced the expression of an anti-oxidative enzyme, heme oxygenase 1, and its upstream transcription factor, nuclear factor E2-related factor-2, in a dose-dependent manner. On the other hand, MMEE was associated with a reduction in NO production, which was induced by the lipopolysaccharide treatment of RAW 264.7 cells. The expression of inducible nitric oxide synthase, which is the upstream regulator of NO production, was also inhibited. Taken together, these results suggest that MMEE has anti-oxidative and anti-inflammatory properties, thus appearing to be a potential anti-oxidant and anti-inflammatory agent. The further identification of active compounds that confer the biological activities of MMEE may be necessary.

A Study on Nutritional Status of Iron in Korean Normal and Obese Male Elementary School Students (정상 남자 아동과 비만 남자 아동의 철 영양상태에 관한 연구)

  • Lee, Seung-A;Bae, Yun-Jeong;Seung, Jeong-Ja
    • Journal of the Korean Dietetic Association
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    • v.12 no.1
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    • pp.55-67
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    • 2006
  • The purpose of this study was to compare the food behavior and nutrient intakes, and the analysis of constituent parts of blood related to iron nutrition conditions and to investigate the correlation between iron nutrition conditions and obesity. Each subject was assigned to one of such as normal(n=55) and obese groups(n=60) according to their obesity index. Their nutritional status, serum iron status were evaluated based in 24-hr dietary recalls, questionnaire and blood analysis. The mean age of the normal group(10.76 years) and that of the obese group(10.95 years) made little difference. The height(p<0.05), weight(p<0.001), and obesity index(p<0.001) in the obese group were higher than those of the normal group in every respect. The 63.55% of the subject of this study gave an answer 'They eat breakfast daily', and the frequency of eating breakfast made little difference. The average intakes of energy were 1669.44 kcal(84.19% of EER) in the obese group and 1673.59 kcal(86.72% of EER) in the normal group respectively, which made little difference between two groups. The intake of nutrients such as calcium and folate was as insufficient as below 75% of RI. The heme iron intake of the obese group was low that of the normal group(p<0.05). Among the major 20 iron resources the iron intake through animal food intake were 1.43 mg(12.00%) in the obese group and 1.93 mg(17.72%) in the normal group respectively. The rating of iron condition in the blood showed that the number of red blood cell(p<0.01), the hemoglobin(p<0.001), and hematocrit(p<0.01) of the obese group were lower than those of the normal group. These results suggest that there should be the right choice of food according to its bioavailability and nutritive guidance to appropriate food behavior, in addition to increasing iron intake to improve iron nutrition conditions of obese male elementary school students.

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