• Title/Summary/Keyword: Induction expression

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Effects of Sargassumpallidum on 2,4,6-Trinitrobenzene Sulfonic Acid-Induced Colitis in Mice (해조가 2,4,6-trinitrobenzene-sulfonic acid로 유발된 염증성 장질환 동물모델에 미치는 영향)

  • Lee, Sang-Wook;Ryu, Bong-Ha;Park, Jae-Woo
    • The Journal of Internal Korean Medicine
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    • v.31 no.2
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    • pp.224-241
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    • 2010
  • Objectives : The aim of the current study was to investigate the effects of Sargassum (Sargassum pallidum (TURN.) C. AG.; SP) on the experimental colitis induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS) in mice. Methods : ICR mice were divided into 7 groups (NOR, CON, $SS50\times5$, $SP20\times3$, $SP50\times3$, $SP20\times5$, $SP50\times5$). TNBS processing was intrarectally applied to all experimental groups on the 3rd experiment day, except the normal group (NOR). For investigating the prophylactic effect, SP at doses of 20 mg/kg ($SP20\times5$) and 50 mg/kg ($SP50\times5$) were orally administered for 5 days. The SP at doses of 20 mg/kg ($SP20\times3$) and 50 mg/kg ($SP50\times3$) were orally administered for 3 days after the colitis induction in order to check the effect of treatment. As a positive control group, sulfasalazine 50 mg/kg ($SS50\times5$) was administrated. Macroscopic findings of epithelial tissue on mice were measured by colon length and macroscopic score. Histologic findings were also checked by crypt cell, epithelial cell, inflammatory cell and edema of submucosa. We measured the ability of SP to inhibit lipid peroxidation and myeloperoxidase activity. We also measured levels of the inflammatory markers, interleukin (IL)-$1\beta$ and cyclooxygenase-2 (COX-2), its transcription factor activation, phospho-NF-${\kappa}B$ (pp65), in the colon by enzyme-linked immunosorbent assay and immunoblot analysis. We measured activation of fecal bacterial enzyme, $\beta$-glucuronidase and degradation activation of fecal glycosaminoglycan (GAG), and hyaluronic acid. Results : Oral administration of SP on mice inhibited TNBS-induced colon shortening and myeloperoxidase activity in the colon of mice as well as IL-$1\beta$ and COX-2 expression. SP also inhibited TNBS-induced lipid peroxidation and pp65 activation in the colon of mice. SP inhibited $\beta$-glucuronidase activation and fecal hyaluronic acid degradation activation as well. Conclusions : SP could be a possible herbal candidate and preventive prebiotic agent for treating inflammatory bowel disease (IBD). Further experiments to differentiate effects of SP on IBD, such as other solutions and extracting times, might be promising.

The Change of c-jun Promoter Activity in TPA-Induced U937 Cells Infected with Human Cytomegalovirus (HCMV) (TPA로 분화된 U937 세포에서 사람 세포거대바이러스에 의한 c-jun Promoter 활성도의 변화)

  • Park, Chung-Gyu;Kim, Dae-Joong;Kim, Jin-Hee;Han, Tae-Hee;Hwan, Eung-Soo;Choi, Myong-Sik;Kook, Yoon-Hoh;Choi, Sung-Bae;Cha, Chang-Yong
    • The Journal of Korean Society of Virology
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    • v.29 no.2
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    • pp.129-136
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    • 1999
  • Transient transfection assay has been done to evaluate whether the c-jun activation would be prerequisite to the induction of permissiveness against human cytomegalovirus using in vitro cell model in which U937 has been induced to express CD11b and CD14 to become potential monocyte/macrophage cells by TPA treatment. U937 cells were treated with $10\;{\mu}M$, $50\;{\mu}M$ or $100\;{\mu}M$ of TPA. The cell morphology change was observed and the expression of the CD11b and CD14 was confirmed by FACS. Differentiated cells were transfected with pJLuc reporter vector which contained the wild type murine c-jun promoter spanning the SP1, CTF, ATF/CREB and MEF-2 binding sites upstream of the firefly luciferase gene. After 48 hrs of transfection, the cells were infected with HCMV Towne strain and the luciferase activity was assessed at 1 hand 4 h pi. The transfection assay showed no activation of the c-jun promoter at 1 h pi, instead, it showed 2 times increase of the its activity at 4 h pi. There was no difference of the c-jun promoter activation between TPA treated and untreated U937 cells, implying that c-jun activation might not be prerequisite for allowing cells to be premissive to HCMV, although HCMV infection itself could activate c-jun promoter.

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Tolerance to Potato Soft Rot Disease in Transgenic Potato Expressing Soybean Ferritin Gene (대두 철분결합단백질 유전자 발현 형질전환 감자의 감자무름병 방어 증진효과)

  • Bae, Shin-Chul;Yeo, Yun-Soo;Heu, Sung-Gi;Hwang, Duk-Ju;Byun, Myung-Ok;Go, Seung-Joo
    • Journal of Plant Biotechnology
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    • v.29 no.4
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    • pp.229-233
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    • 2002
  • Ferritin is ubiquitous in bacteria, animals and plants. Ferritin is thought to play two main roles in living cells to provide iron for the synthesis of iron protein such as ferretoxin and cytochromes and to prevent damage from radicals produced by iron/dioxygen interaction. To enhance the resistance of potato to Erwinia carotovora, the soybean ferritin gene was introduced into the potato either under CaMV 35S or hsr203J promoter. Potato transgenic plants were screened by PCR analysis using specific primers to the ferritin gene. Expression of ferritin gene under CaMV 35S and hsr203J promoter in potato transgenic plants was confirmed by northern blot analysis. hsr203J promoter known to pathogen inducible in tobacco drives the induction upon Phytophthora infestan in potato and the transcript level of ferritin gene was extremely high after 24 hours post inoculation. One of transformants under CaMV 35S promoter was increased 2.5 fold than untransformant. Each one of transgenic potato containing gene promoter CaMV 35S and hsr203J-ferrtin fusion exhibited tolerance against potato soft rot.

Induction of Apaopotis by Water Extract of Cordyceps militaris (WECM) in Human Hepatocellular Carcinoma HepG2 Cells. (동충하초 열수 추출물에 의한 인체 간암세포 성장억제 및 apoptosis 유발에 관한 연구)

  • Kim, Kyung-Mi;Park, Cheol;Choi, Yung-Hyun;Lee, Won-Ho
    • Journal of Life Science
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    • v.18 no.6
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    • pp.804-813
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    • 2008
  • Cordyceps militaris, the Chinese medicinal fungal genus Cordyceps, is reported to possess many pharmacological activities including immunological stimulating, anti-cancer, anti-virus and anti-infection activities. However, the molecular mechanisms of C. militaris on biochemical actions in cancer have not been clearly elucidated yet. In the present study, we investigated the anti-proliferative activity of the water extract of C. militaris (WECM) in human hepatocellular carcinoma HepG2 cells. It was found that WECM could inhibit the cell growth in a dose-dependent manner, which was associated with morphological changes and apoptotic cell death such as formation of apoptotic bodies and increased populations of apoptotic sub-G1 phase. Apoptotic cell death of HepG2 cells by WECM was connected with a up-regulation of pro-apoptotic Bax expression, tumor suppressor p53 and cyclin-dependent kinase inhibitor p21 (WAF1/CIP1). In addition, WECM treatment induced the proteolytic activation of caspase-3 and a concomitant degradation and/or inhibition of poly (ADP-ribose) polymerase (PARP), ${\beta}-catenin$ and phospholipase $(PLC)-{\gamma}1$ protein. Furthermore, caspase-3 inhibitor, z-DEVD-fmk, significantly inhibited WECM-induced apoptosis demonstrating the important role of caspase-3 in the observed cytotoxic effect. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of C. militaris.

Roles of TLR-4 and NF-κB in Interleukin-6 Expression Induced by Heat Shock Protein 90 in Vascular Smooth Muscle Cells (혈관평활근세포에서 HSP90에 의한 IL-6 발현에 TLR-4와 NF-κB의 작용)

  • Rhim, Byung-Yong;Kim, Kang-Seong;Kim, Koan-Hoi
    • Journal of Life Science
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    • v.18 no.12
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    • pp.1637-1643
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    • 2008
  • This study has investigated whether extracellular HSP90 predisposes vascular smooth muscle cells (VSMCs) to pro-inflammatory phenotype. Exposure of rat aortic smooth muscle cells to HSP90 not only enhanced IL-6 release but also profoundly induced IL-6 transcript via promoter activation. HSP90-induced IL-6 promoter activation was suppressed by dominant-negative forms of Toll-like receptor (TLR)-4 and myeloid differentiation factor 88 (MyD88), but not by dominant-negative-forms of TLR-3 and TIR-domain-containing adapter-inducing interferon-${\beta}$ (TRIF). Curcumin, which inhibits dimerization of TLR-4, also attenuated the IL-6 induction by HSP90. Mutation at the NF-${\kappa}B$- or C/EBP-binding site in the IL-6 promoter region suppressed the promoter activation in response to HSP90. The gene delivery of $I{\kappa}B$ using recombinant adenoviruses and treatment with resveratrol, which inhibit NF-${\kappa}B$ activity, attenuated the HSP90-induced IL-6 release from VSMCs. The present study proposes that extracellular HSP90 would contribute to inflammatory reaction in the stressed vasculature by inducing IL-6 in VSMCs, and that TLR-4 and NF-${\kappa}B$ would play active roles in the process.

Isolation of CONSTANS as a TGA4/OBF4 Interacting Protein

  • Song, Young Hun;Song, Na Young;Shin, Su Young;Kim, Hye Jin;Yun, Dae-Jin;Lim, Chae Oh;Lee, Sang Yeol;Kang, Kyu Young;Hong, Jong Chan
    • Molecules and Cells
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    • v.25 no.4
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    • pp.559-565
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    • 2008
  • Members of the TGA family of basic domain/leucine zipper transcription factors regulate defense genes through physical interaction with NON-EXPRESSOR OF PR1 (NPR1). Of the seven TGA family members, TGA4/octopine synthase (ocs)-element-binding factor 4 (OBF4) is the least understood. Here we present evidence for a novel function of OBF4 as a regulator of flowering. We identified CONSTANS (CO), a positive regulator of floral induction, as an OBF4-interacting protein, in a yeast two-hybrid library screen. OBF4 interacts with the B-box region of CO. The abundance of OBF4 mRNA cycles with a 24 h rhythm under both long-day (LD) and short-day (SD) conditions, with significantly higher levels during the night than during the day. Electrophoretic mobility shift assays revealed that OBF4 binds to the promoter of the FLOWERING LOCUS T (FT) gene, a direct target of CO. We also found that, like CO and FT, an OBF4:GUS construct was prominently expressed in the vascular tissues of leaf, indicating that OBF4 can regulate FT expression through the formation of a protein complex with CO. Taken together, our results suggest that OBF4 may act as a link between defense responses and flowering.

Anti-inflammatory Constituents of the Aerial Parts of Trichosanthes kirilowii in BV2 Microglial Cells (괄루경엽의 BV2 미세아교세포에서의 항염증 활성 성분)

  • Li, Xiao Jun;Kim, Kwan-Woo;Ko, Wonmin;Kim, Dong-Cheol;Yoon, Chi-Su;Liu, Xiang Qian;Kim, Jong-Su;Jang, Kyu-Kwan;Kang, Dae-Gil;Lee, Ho-Sub;Oh, Hyuncheol;Kim, Youn-Chul
    • Korean Journal of Pharmacognosy
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    • v.47 no.1
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    • pp.7-11
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    • 2016
  • The aerial part of Trichosanthes kirilowii Maxim. (Cucurbitaceae), has long been used in traditional Korean and Chinese medicines for the treatment of heatstroke. We isolated and identified three flavones, luteolin-7-O-${\beta}$-D-glucopyranoside(1), luteolin-4'-O-${\beta}$-D-glucopyranoside(2), luteolin(3) from its methanolic extract. In the present study, we found that luteolin attenuates the lipopolysaccharide(LPS)-induced inflammation in BV2 microglial cells. Luteolin significantly inhibited LPS-induced production of pro-inflammatory mediators such as nitric oxide(NO) and prostaglandin $E_2(PGE_2)$ in BV2 microglia in a concentration-dependent manner without cytotoxic effect. Luteolin dose-dependently suppressed the protein expression of inducible nitric oxide synthase(iNOS) and cyclooxygenase-2(COX-2). In addition, luteolin also showed significant induction of heme oxygenase(HO)-1. These results suggest that both the aerial part of T. kirilowii and luteolin may be good candidates to regulate LPS-induced inflammatory response.

Molecular Cloning and Characterization of Salt-inducible Aldolase from Salicornia herbacea (퉁퉁마디로부터 염에 의하여 유도되는 Aldolase 유전자의 분리 및 발현분석)

  • Cha, Joon-Yung;Netty Ermawati;Kim, Soon-Gil;Lee, Jeung-Joo;Lim, Chae-Oh;Chung, Woo-Sik;Lee, Kon-Ho;Son, Dae-Young
    • Journal of Plant Biotechnology
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    • v.30 no.4
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    • pp.323-328
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    • 2003
  • Soil salinity is one of the most serious abiotic stresses limiting the productivity of agricultural crops. To cope with salt stress, plants respond with physiological, developmental and biochemical changes, including the synthesis of a number of proteins and the induction of gene expression. Salicornia herbacea is a halophytic plant that grows in salt marshes and on muddy seashores. In order to understand the biochemical and molecular mechanisms of salt tolerance in S. herbacea, we isolated several genes that involved in the salt tolerance by mRNA differential display. Here we report the cloning of a cDNA encoding fructose-1, 6-bisphosphate aldolase, named ShADL, which is 1293 bp long and contains an open reading frame consisted of 359 amino acids with calculated molecular mass of 39 kDa. ShADL protein showed 86% identity with Arabidopsis and 78% with aldolase of common ice plant. Northern blot analysis revealed that the transcript of ShADL gene was increased dramatically depending on the NaCl concentrations.

In vitro Anti-diabetic Effects of Crude Extracts of Platycodi Radix (In vitro에서 길경 추출 분획물의 항당뇨 효과 조사)

  • Ko, Byoung-Seob;Kwon, Dae-Young;Hong, Sang-Mee;Park, Sun-Min
    • Korean Journal of Food Science and Technology
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    • v.39 no.6
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    • pp.701-707
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    • 2007
  • Anti-diabetic effect of Platycodi radix (PR) extract fractions was determined if vitro by investigating insulin-like action, insulin sensitizing action, glucose-stimulated insulin secretion, gene expression related to ${\beta}-cell$ function and mass, and ${\alpha}$-glucoamylase suppressing action. Insulin-like activity was not promoted by the treatment of PR methanol factions in 373-L1 fibroblast. However, treatment with 0, 20 and 100% PR methanol fractions along with 1 ng/mL insulin increased insulin-stimulated glucose uptake in 373-L1 adipocytes. In addition, the treatment of 0% and 100% methanol fractions along with differentiation inducers significantly increased the differentiation of 373-L1 fibroblasts to adipocytes. These fractions may contain insulin sensitizer. The 20%, 80% and 100% methanol fractions enhanced glucose-stimulated insulin secretion in Min6 cells, insulin secreting cell line. This was related to the mechanism to promote glucose sensing and ${\beta}-cell$ proliferation, which was regulated by the induction of IRS-2, glucokinase and PDX-1 genes. As expected, 20, 80 and 100% methanol fractions increased mRNA levels of IRS-2, glucokinase and PDX-1 genes. However, PR fractions did not affect the ${\alpha}-glucoamylase$ activity in vitro. These data suggested that PR extract fractions have anti-diabetic actions through improving insulin sensitization, glucose-stimulated insulin secretion, and ${\beta}-cell$ proliferation. Therefore, PR extracts can be beneficial for anti-diabetic treatment in lean diabetic patients.

Protective Effects of Water Extracts Composed of Adenophora triphylla var. japonica Hara on the Acetaminophen-induced Hepatotoxicity (아세트아미노펜에 의해 유도된 간독성 모델에서 잔대를 주원료로 하는 추출물의 간 보호 효과)

  • Gum, Sang-Il;Lee, Dong-Ung;Cho, Min-Kyung
    • Korean Journal of Food Science and Technology
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    • v.39 no.6
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    • pp.688-693
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    • 2007
  • The hepatoprotective effects of water extracts composed of Adenophora triphylla var japonica Hara (ATJH) on acetaminophen (APAP)-induced hepatotoxicity were investigated in vivo and in vivo. The effects of ATJH on liver toxicity induced by APAP were assessed by blood biochemical and histopathological analyses. APAP treatment (350 mg/kg) caused severe liver injury in mice as indicated by their significantly elevated plasma aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels. Pretreatment with ATJH for 3 or 7 days attenuated the increases in ALT and AST when challenged with APAP. The reductions in viability caused by high dose of APAP (450 mg/kg) in vivo were reversed by pretreatment with ATJH. These protective effects of ATJH against APAP-induced toxicity were consistent with the results from the histopathological examinations. We next examined the effects of ATJH on the gene expression of glutathione S-transferases (GSTs) that detoxify the metabolic intermediates of APAP in H4IIE cells. The hepatic GST protein levels [$\alpha$ class (GSTA2, GSTA3/5)] were significantly elevated in a dose-dependent manner by ATJH treatment. In summary, ATJH is effective at protecting against APAP-induced hepatotoxicity by GST induction, implying that ATJH should be considered a potential chemopreventive agent.