• Title/Summary/Keyword: rho-O cells

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Inhibitory Effects of Natural Plant Extracts on ICAM-1/LFA-1 Mediated Adhesion of HL-60 Cells (자생식물 추출물의 세포접착인자 저해활성 검색)

  • Kwon, Oh-Eok;Lee, Seung-Woong;Chung, Mi-Yeon;Kim, Young-Ho;Lee, Hyun-Sun;Kim, Young-Kook;Rho, Mun-Chual
    • Korean Journal of Pharmacognosy
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    • v.33 no.4 s.131
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    • pp.343-351
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    • 2002
  • Atherosclerosis is a progressive disease characterized by the accumulation of lipids and fibrous elements in the arteries. Monocyter/macrophages are involved in many aspects of the development of atherosclerotic plaques. It is known that the intercellular adhesion molecule-1(ICAM-1) expressed preferentially on endothelial cells of atherosclerotic plaque, promotes local adhesion and transendothelial migration of monocytes, neutrophils, and lymphocytes. Using the human promyelocytic leukemia HL-60 cell line, we investigated the inhibitory effects of methanol extracts of 175 natural plants on ICAM-1/LFA-1 mediated cell adhesion. Eight kinds of methanol extracts of tested plants inhibited PMA-induces homotypic aggregationof HL-60 cells without cytotoxicity at the concentration of $6.25\;{\mu}g/ml$. They were divided two fractions of $CHCI_3$ and $H_2O$ to use solvent partition. Among them, $CHCI_3$ extract $(1.0\;{\mu}g/ml)$ of Saururus chinensis and Chloranthus japonicus singificantly inhibited aggregation of HL-60 cells without cytotoxicity, respectively.

Subpopulations of miniature pig mesenchymal stromal cells with different differentiation potentials differ in the expression of octamer-binding transcription factor 4 and sex determining region Y-box 2

  • Jeon, Ryounghoon;Park, Sungjo;Lee, Sung-Lim;Rho, Gyu-Jin
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.3
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    • pp.515-524
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    • 2020
  • Objective: Human mesenchymal stromal cells (MSCs) exhibit variable differentiation potential and can be divided accordingly into distinct subpopulations whose ratios vary with donor age. However, it is unknown whether the same is true in pigs. This study investigated MSC subpopulations in miniature pig and compared their characteristics in young (2 to 3 months) and adult (27 to 35 months) pigs. Methods: Osteogenic, chondrogenic, and adipogenic capacity of isolated MSCs was evaluated by von Kossa, Alcian blue, and oil red O staining, respectively. Cell surface antigen expression was determined by flow cytometry. Proliferative capacity was assessed with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Expression of marker genes was detected by quantitative real-time polymerase chain reaction. Results: Porcine MSCs comprised cells with trilineage and bilineage differentiation potential (tMSCs and bMSCs, respectively) and non-differentiating stromal cells (NDSCs). The tMSC and bMSC fractions were smaller in adult than in young pigs (63.0% vs 71.2% and 11.6% vs 24.0%, respectively, p<0.05); NDSCs showed the opposite trend (25.4% vs 4.8%; p<0.05). Subpopulations showed no differences in morphology, cell surface antigen expression, or proliferative capacity, but octamer-binding transcription factor 4 (OCT4) expression was higher in tMSCs than in bMSCs and NDSCs (p<0.05), whereas sex determining region Y-box 2 (SOX2) expression was higher in tMSCs and bMSCs than in NDSCs (p<0.05). Aging had no effect on these trends. Conclusion: Porcine MSCs comprise distinct subpopulations that differ in their differentiation potential and OCT4 and SOX2 expression. Aging does not affect the characteristics of each subpopulation but alters their ratios.

Antioxidant activity of white ginseng extracts prepared by enzyme treatment on V79-4 cells induced by oxidative stress (효소처리에 의한 백삼 저분자 화합물의 V79-4 세포주에 대한 항산화 활성)

  • Kim, Young-Chan;Yim, Joo-Hyuk;Rho, Jeong-Hae;Cho, Chang-Won;Rhee, Young-Kyung
    • Journal of Ginseng Research
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    • v.31 no.4
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    • pp.203-209
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    • 2007
  • This study examined the extraction yields, total phenolic compounds content and the antioxidant activities on V79-4 cells of white ginseng extracts prepared by enzyme treatment. Yields of crude extract were 29.5-76%, and total phenolic compounds content showed 0.45-2.2% according to enzyme treatments. Pectinase treatment group showed the highest values of extraction yields and total phenolic compounds content. Pectinase and a-amylase treatment groups protected V79-4 cell viability(above 50%) against $H_2O_2$-induced oxidative damage. In the result of antioxidant enzyme activity evaluation in cells, enzyme treatments did not show the significant difference of SOD activity (p>0.05). However, pectinase treatment group exhibited increased CAT and GPx activities (p>0.05). Also, pectinase and protease treatment group inhibited MDA formation (>50%) in the lipid peroxidation protection experiment.

Effect of Moutan Cortex Radicis on Gene Expression Profile of Differentiated PC12 Rat Cells Oxidative-stressed with Hydrogen Peroxide (모단피의 PC12 cell 항산화 효과와 관련 HO, MIF, COMT 유전자 발현에 미치는 영향)

  • Son Mu Song;Rho Sam Woong;Ko Eun Jung;Na Youn Gin;Bae Hyun Su;Hong Moo Chang;Shin Min Kyu
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.4
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    • pp.905-913
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    • 2003
  • Oriental medicine explains aging as the weakening of Kidney-ai, and Kidney-strengthening herbal medicines such as Yukmijihwang-tang have been studied for anti-aging effects. In Western Medicine, the hypothesis that reactive oxidant species(ROS) contribute to the aging process is generally accepted. It has been reported that Moutan Cortex Radicis extract (MCR) was the most effective constituent of Yukmijihwang-tang in decreasing ROS production in oxidative-stressed cells. The purpose of this study is to confirm the anti-oxidant effect of MCR on PC12 cells, the expression of Heme oxygenase (HO), Macrophage migradon inhibitory factor (MIF), Catechol-O-methyltransferase (COMT) using real time RT PCR. PC12 cells were treated without or with hydrogen peroxide in the presence or absence of MCR using MTS assay. Hydrogen peroxide decreased the viability of PC12 cells by 53% and MCR did not influence that of stressed PC 12 cells irrespective of dose or incubation period. However, MCR showed an inhibitory effect on production of ROS in stressed cells, both dose and incubation time dependently. In particular, 1 ㎎/㎖ of MCR for 24 h culture almost returned to normal level. In the quantiation of anti-aging related gene expression, MCR at 1 ㎎/㎖ increased the expression of HO by 370%, MIF by 180% and COMT by 280% through real time RT PCR. In conclusion, MCR treatment protected PC12 cells from hydrogen peroxide and decreased ROS production and enhanced anti-oxidative gene expression such as HO, COMT and MIF, which suggests that MCR is involved in controlling anti-aging of nerve cells through elimination of cytotoxic stimuli.

Effect of Moutan Cortex Radicis on gene expression profile of differentiated PC12 rat cells oxidative-stressed with hydrogen peroxide (모단피의 PC12 cell 산화억제 효과 및 neuronal 유전자 발현 profile 분석에 대한 연구)

  • Kim Hyun Hee;Rho Sam Woong;Na Youn Gin;Bae Hyun Su;Shin Min Kyu;Kim Chung Suk;Hong Moo Chang
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.529-541
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    • 2003
  • Yukmijihwang-tang has been widely used as an and-aging herbal medicine for hundred years in Asian countries. Numerous studies show that Yukmijihwangtang has anti-oxidative effect both in vivo and in vitro. It has been reported that Moutan Cortex Radicis extract (MCR) was the most effective herb in Yukmijihwang-tang on undifferentiated PC12 cells upon oxidative-stressed with hydrogen peroxide. The purpose of this study is to; 1) evaluate the recovery of neuronal damage by assessing the anti-oxidant effect of MCR on PC12 cells differentiated with nerve growth factor (NGF), 2) identify candidate genes responsible for anti-oxidative effect on differentiated PC12 cells by oligonucleotide chip microarray. PC12 cells, which were differentiated by treating with NGF, were treated without or with hydrogen peroxide in the presence or absence of various concentration of MCR. Cell survival was determined by using MTS assay. Measurement of intracellular reactive oxygen species (ROS) generation was determined using the H2DCFDA assay The viability of cells treated with MCR was significantly recovered from stressed PC12 cell. In addition, wide rage of concentrations of MCR shows dose-dependent inhibitory effect on ROS production in oxidative-stressed cells. Total RNAs of cells without treatment(Control group), only treated with H₂O₂ (stressed group) and treated with both H₂O₂ and of MCR (MCR group) were isolated, and cDNAs was synthesized using oligoT7(dT) primer. The fragmented cRNAs, synthesized from cDNAs, were applied to Affymetrix GeneChip Rat Neurobiology U34 Array. mRNA of Calcium/calmodulin-dependent protein kinase II delta subunit(CaMKII), neuron glucose transporter (GLUT3) and myelin/oligodendrocyte glycoprotein(MOG) were downregulated in Stressed group comparing to Control group. P2X2-5 receptor (P2X2R-5), P2X2-4 receptor (P2X2R-4), c-fos, 25 kDa synaptosomal attachment protein(SNAP-25a) and GLUT3 were downregulated, whereas A2 adenosine receptor (A2AR), cathechol-O-methyltransferase(COMT), glucose transporter 1 (GLUT1), EST223333, heme oxygenase (HO), VGF, UI-R-CO-ja-a-07-0-Ul.s1 and macrophage migration inhibitory factor (MIF) were upregulated in MCA group comparing to Control group. Expression of Putative potassium channel subunit protein (ACK4), P2X2A-5, P2X2A-4, Interferon-gamma inducing factor isoform alpha precursor (IL-18α), EST199031, P2XR, P2X2 purinoceptor isoform e (P2X2R-e), Precursor interleukin 18 (IL-18) were downregulated, whereas MOO, EST223333, GLUT-1, MIF, Neuronatin alpha, UI-R-C0-ja-a-07-0-Ul.s1, A2. adenosine receptor, COMT, neuron-specific enolase (NSE), HO, VGF, A rat novel protein which is expressed with nerve injury (E12625) were upregulated in MCR group comparing to Stressed group. The results suggest that decreased viability and AOS production of PC12 cell by H₂O₂ may be, at lease, mediated by impaired glucose transporter expression. It is implicated that the MCR treatment protect PC12 cell from oxidative stress via following mechanisms; improving glucose transport into the cell, enhancing expression of anti-oxidative genes and protecting from dopamine cytotoxicity by increment of COMT and MIF expression. The list of differentially expressed genes may implicate further insight on the action and mechanism behind the anti-oxidative effects of herbal extract Moutan Cortex Radicis.

Metabolic Engineering for Resveratrol Derivative Biosynthesis in Escherichia coli

  • Jeong, Yu Jeong;Woo, Su Gyeong;An, Chul Han;Jeong, Hyung Jae;Hong, Young-Soo;Kim, Young-Min;Ryu, Young Bae;Rho, Mun-Chual;Lee, Woo Song;Kim, Cha Young
    • Molecules and Cells
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    • v.38 no.4
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    • pp.318-326
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    • 2015
  • We previously reported that the SbROMT3syn recombinant protein catalyzes the production of the methylated resveratrol derivatives pinostilbene and pterostilbene by methylating substrate resveratrol in recombinant E. coli. To further study the production of stilbene compounds in E. coli by the expression of enzymes involved in stilbene biosynthesis, we isolated three stilbene synthase (STS) genes from rhubarb, peanut, and grape as well as two resveratrol O-methyltransferase (ROMT) genes from grape and sorghum. The ability of RpSTS to produce resveratrol in recombinant E. coli was compared with other AhSTS and VrSTS genes. Out of three STS, only AhSTS was able to produce resveratrol from p-coumaric acid. Thus, to improve the solubility of RpSTS, VrROMT, and SbROMT3 in E. coli, we synthesized the RpSTS, VrROMT and SbROMT3 genes following codon-optimization and expressed one or both genes together with the cinnamate/4-coumarate:coenzyme A ligase (CCL) gene from Streptomyces coelicolor. Our HPLC and LC-MS analyses showed that recombinant E. coli expressing both ScCCL and RpSTSsyn led to the production of resveratrol when p-coumaric acid was used as the precursor. In addition, incorporation of SbROMT3syn in recombinant E. coli cells produced resveratrol and its mono-methylated derivative, pinostilbene, as the major products from p-coumaric acid. However, very small amounts of pterostilbene were only detectable in the recombinant E. coli cells expressing the ScCCL, RpSTSsyn and SbROMT3syn genes. These results suggest that RpSTSsyn exhibits an enhanced enzyme activity to produce resveratrol and SbROMT3syn catalyzes the methylation of resveratrol to produce pinostilbene in E. coli cells.

Preventive Effect of Crude Drug Preparation (E-kong-san) on Cisplatin induced Nephrotoxicity (생약제제인 이공산(異功散)의 Cisplatin 유도 신장독성 보호 효과)

  • Rho, Young-Soo;Ahn, Kyoo-Seok;Chang, Sung-Goo;Jung, Jee-Chang;Lee, Kyung-Tae
    • Korean Journal of Pharmacognosy
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    • v.29 no.3
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    • pp.258-264
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    • 1998
  • Nephroprotective effects of a crude drug-preparation (Ekongsan) were determined from cisplatin induced renal injury in vivo and in vitro. Ekongsan decreased cisplatin induced the cytotoxicity on rabbit kidney proximal tubule and human renal cortical cells by MTT assays and sustained glucose consumption on cisplatin-induced human renal cortical tissue. Levels of creatinine and blood urea nitrogen (BUN) in serum after administration of cisplatin (0.75 mg/kg, i.p.) to Ekongsan (0.75 g/kg/d, p.o.) pretreated rats were markedly lower compared to those of cisplatin-treated rats. Moreover, the administration of Ekongsan significantly inhibited the loss of body weight of cisplatin-injected rats. These findings suggest that Ekongsan is an active prescription in protection against nephrotoxicity of cisplatin.

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PHENOL DERIVATIVES EFFECTS ON GLUTAMIC ACID FERMENTATION (Phenol 유도체 처리가 Glutamin산 생성균의 발효증가에 미치는 영향에 대하여)

  • RHO Yung Jae;LEE Kyung Hee
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.12 no.2
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    • pp.95-102
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    • 1979
  • Brevibacterium flavum treated with phenol derivatives, guaiacol and o-vanillin has been revealed the marked increased ability in glutamic acid fermentation as 14.2 g/l in o-vanillin treated, 12.5 g/l in guaiacol treated while the 7.0 g/1 in nontreated cell. The increased ability of phenol derivatives treated cells in glutamic acid fermentation was ascribed to the formation of charge-transfer complex between phenols and oxygen. The charge-transfer complex effectively supply the oxygen to the fermention system in spite of high potential gradient in oxygen transfer formed by high cell concentration as insulator on film of air-liquid interface.

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Imaging of Tumor Proliferation Using Iodine-131-Iodomethyluridine (Iodine-131-Iodomethyluridine을 이용한 종양세포증식의 영상화에 관한 실험적 연구)

  • Min Kyung-Yoon;Kim, Chang-Guhn;Kim, Hyun-Jeong;Lim, Hyung-Guhn;Rho, Ji-Young;Juhng Seon-Kwan;Won Jong-Jin;Yang, David J.
    • The Korean Journal of Nuclear Medicine
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    • v.30 no.3
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    • pp.344-350
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    • 1996
  • Purpose : Noninvasive imaging of tumor cell proliferation could be helpful in the evaluation of tumor growth potential and could provide an early assessment of treatment response. Radiolabeled thymidine, uridine and adenosine have been used to evaluate tumor cell proliferation. These nucleoside analogs are incorporated into DNA during proliferation. Iodine-131-Iodomethyluridine, an analog of Iodine-131-Iododeoxyuridine, is also involved in DNA/RNA synthesis. The purpose of this study was to develop Iodine-131-Iodomethylurdine and image tumor proliferation using Iodine-131-Iodomethyluridine. Materials and Methods : Radiosynthesis of Iodine-131-5-Iodo-2'-O-methyluridine (Iodine-131-Iodomethyluridine) was prepared from 10 mg of 2'-O-methyluridine(Sigma chemical Co., St. Louis, Missouri) and 2.1 mCi(SP. 10Ci/mg) of Iodine-131-labeled sodium iodide in $100{\mu}l$ of water using iodogen reaction. Female Fischer 344 rats were inoculated in the thigh area with breast tumor cells(13765 NF, $10^5$ cells/rat S.C.). After 14 days, the Iodine-131-Iodomethyluridine $10{\mu}Ci$ was injected to three groups of rats(3/group). The percent of injected dose per gram of tissue weight was determined at 0.5-hours, 2-hours, 4-hours, and 24-hours respectively. Tumor bearing rats after receiving Iodine-131-Iodomethyluridine($50{\mu}Ci$ IV) were euthanized at 2 hours after injection. Autoradiography was done using freeze-dried $50{\mu}m$ coronal section. After injection of Iodine-131- Iodomethyluridine ($10{\mu}Ci$/rat, IV) in three breast tumor-bearing rats, planar scintigraphy was taken at 45 minutes, 90 minutes and 24 hours. Results : Iodine-131-Iodomethyluridine was conveniently synthesized using iodogen reaction. The biodistribution showed fast blood clearance and the tumor-to-tissue uptake ratios showed that optimal imaging time was at 2 hours postinjection. Autoradiogram and planar scintigram indicated that tumor could be well visualized. Conclusion : The findings suggest that Iodine-131-Iodomethyluridine, a new radio-iodinated nucleoside, has potential use for evaluation of active regions of tumor growth.

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Differential Cytotoxicity of Penta-O-galloyl-β-D-glucose in Human Cancer and Normal Cell Lines of Various Origins (사람의 다양한 조직에서 기원하는 암세포 및 정상세포에 대한 penta-O-galloyl-β-D-glucose의 세포독성 효과)

  • Lee, Hyeon-Jeong;Kim, Min-Gyeong;Lee, Song-Yeong;Song, Min-Hyock;Kim, Yoon-Dong;Ha, Jeong-Sook;Jeong, Gie-Joon;Rho, Gyu-Jin;Jeon, Byeong-Gyun
    • Journal of Life Science
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    • v.26 no.11
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    • pp.1320-1329
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    • 2016
  • The present study examined the cytotoxic effects of 1, 2, 3, 4, 6-penta-O-galloyl-${\beta}$-D-glucose (PGG), known as the pentahydroxy gallic acid ester of glucose, in the various human cancer cell lines (A-549, MDA-MB-231, U87-MG, MCF-7 and PANC-1), normal MRC-5 fetal fibroblasts, and dental papilla tissue- derived mesenchymal stem cells (DPSCs). Significantly (p<0.05) lower half maximal inhibitory concentration ($IC_{50}$) values were observed in the A-549 and MDA-MB-231 cells showing a high proliferation capacity, compared with other cancer and normal cell lines with a relatively low proliferation capacity. The population doubling time (PDT) was significantly (p<0.05) higher in the $10{\mu}M$ PGG-treated cell lines than those of untreated control cell lines. The present study demonstrated that the $IC_{50}$ value increases proportionally to the extending PDT. A high cell number with senescence-associated ${\beta}-galactosidase$ activity was also observed in the $10{\mu}M$ PGG-treated cells compared with those of untreated control cells. Moreover, the level of telomerase activity was significantly (p<0.05) decreased with $10{\mu}M$ PGG treatment, especially in A-549 and MDA-MB-231 cells showing a high proliferation capacity. Based on these observations, PGG could serve as a potent agent for cancer chemotherapy, as its treatment was more effective in cells with a high proliferation capacity.