• Title/Summary/Keyword: Expression Concentration

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Inhibition Effect of Gamisoyo-san on MITF, TRP-1, TRP-2, Tyrosinase mRNA Expression in Melanoma Cells (B16F10) (멜라노마 세포에서 가미소요산(加味逍遙散)의 MITF, TRP-1, TRP-2, Tyrosinase mRNA 발현 억제 효과)

  • Joo, Da-Hye;Lee, Soo-Yeon;Yoo, Dan-Hee;Lee, Jin-Young
    • The Korea Journal of Herbology
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    • v.29 no.6
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    • pp.157-163
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    • 2014
  • Objectives : Gamisoyo-san complex prescription were made with Angelicae Gigantis Radix, Paeoniae Radix, Atractylodes rhizome white, Hoelen, Bupleuri Radix, Moutan Cortex Radicis, Gardeniae Fructus, Zingiberis Rhizoma Crudus, Menthae Herba. The purpose of this study was to research the whitening effect of the extract from Gamisoyo-san, which is one of the used herbal complex prescription. Methods : This study investigated inhibitory effect of Gamisoyo-san in tyrosinase activity. Cell viability were performed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Then, Gamisoyo-san measured reversed-transcription-PCR for mRNA expression using B16F10 mouse melanoma cells. Results : For whitening effects, the tyrosinase inhibition effect of extract was shown to 52.4% at $5,000{\mu}g/m{\ell}$ concentration. The cell viability on B16F10 melanoma cells of Gamisoyo-san extract showed higher than 75% at $1,000{\mu}g/m{\ell}$ concentration. In this study, an experiment was performed by setting the non-toxic concentration range of 50, 150, $250{\mu}g/m{\ell}$. The Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a positive control. The microphthalmia-associated transcription factor (MITF), tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2), tyrosinase mRNA expression inhibitory by reverse transcription-PCR of Gamisoyo-san extract were decreased by 95.3%, 98.8%, 96.3% and 49.5% at $250{\mu}g/m{\ell}$ which the highest concentration. Conclusions : All these findings could verify that whitening effects of Gamisoyo-san extract by tyrosinase inhibitory activity and mRNA expression. The Gamisoyo-san could be used as material for functional cosmetics, such as skin whitening products.

Effect of Banggibongnyeongtang on the immunohistological change in LPS-induced depression rats (방기복령탕(防己茯苓湯)이 백서에서 LPS로 유도된 우울증에서 면역 조직학적 변화에 미치는 효과)

  • Park, Sung jun;Lee, Tae Hee
    • Herbal Formula Science
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    • v.28 no.1
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    • pp.53-62
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    • 2020
  • Objective : This study is accomplished in order to investigate the effect of banggibongnyeongtang on the immunohistological change in LPS-induced depression rats to confirm the histological result of the previous behavioral and biochemical effect. Methods : LPS 5 ㎍ was injected to lateral ventricle and experimental groups were administered BBT intraperitoneally. The concentration of 5-HT in the Medial Prefrontal Cortex, Striatum, Hippocampus, Amygdala was measured by ELISA. IL-1β, TNF-α mRNA and BDNF mRNA expression in the hippocampus was examined by RT-PCR. Result : BBT enhanced 5-HT concentration at all part of brain but no significantly difference at medial prefrontal cortex and striatum. LPS+BBT400 group increased 5-HT concentration significantly than LPS group at hippocampus and amygdala (p<0.05). BBT decreased IL-1β mRNA expression dose dependently but only with significantly decrease in LPS+BBT400 group than LPS group's in Hippocampus (p<0.05). But BBT did not decrease TNF-α mRNA expression significantly in Hippocampus. BBT increased the expression of BDNF mRNA at hippocampus and LPS+BBT400 group significantly increased comparing with LPS group does (p<0.05). Conclusion : It is postulated that the anti-depressant effect of BBT can be validated through the anti-inflammatory effect, 5-HT concentration increase, and the neuro-protective effect mediated by BDNF by combining the results of the previous report about the behavioral and biochemical effect.

Effect of Corydalis Tuber on the inhibition of proliferation of human uterine leiomyoma cell and apoptotic gene expression (현호색(玄胡索)이 자궁근종세포의 증식 억제와 Apoptosis 관련 유전자 발현에 미치는 영향)

  • Lee, Hee-Jae;Baek, Seung-Hee;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.19 no.2
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    • pp.214-225
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    • 2006
  • Purpose : This study was aimed to investigate the inhibitory effect of Corydalis Tuber on the proliferation of human uterine leiomyoma cell and the expression of gene related the mechanism of cell apoptosis. Methods : We counted the number of suvival cells treated with indicated concentration of Corydalis Tuber and investigated cell viability by MTS assay. Furthermore, flow cytometric analyis were used to dissect between necrosis and apoptosis related with cell cycle and then we observed the differential gene expression by western blot analysis. Results : 1) The inhibitory effect on the proliferation of uterine leiomyoma cell treated with Corydalis Tuber was increased in a concentration and time proportional. 2) The result of flow cytometry analysis, subG1 phase arrest related cell apoptosis was not investigated in uterine leiomyoma cell treated Corydalis Tuber but showed G2/M phase prolongation. 3) The gene expression of p27, p21 related cell cycle was increased according to increasing concentration, but p53 was not exchanged. 4) The dephosphorylation of pRb gene were increased dependent on treatment concentration and pro-caspase 3, CDK4 were not exchanged. Conclusion : This study showed that Corydalis Tuber have the inhibitory effect on the proliferation of human uterine leiomyoma cell but the effect was thoughted no relationship with apoptosis. The inhibitory effect was suggested that dephosphorylation of pRb gene induced with increasing p21, p27 prolonged cell division in G2/M phase.

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Effects of Sochungyong-tang on Cytokine Gene Expression in Mouse Alveolar Macrophage (소청용장(小靑龍湯)이 생쥐의 폐(肺) 대식세포(大食細胞) Cytokine 귀전자(遣傳子) 발현에 미치는 영향)

  • Park, In-Gi;Sim, Sung-Young;Byun, Hak-Sung;Kim, Kyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.18 no.3
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    • pp.1-17
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    • 2005
  • In many recent studies, molecular biological methods have been used to investigate the role of cytokines in pathogenesis of lung disease. This Experiment was conducted to investigate the effects of Sochungyong-tang on gene expressions in Mouse Alveolar Macrophage. Fer this purpose, we observed the cytokines ($IL-1{\beta}$, IL-6, IL-10, iNOS, $MIP-1{\alpha},\;MIP-1{\beta},\;MIP-1{\gamma},\;TGF-{\beta},\;TNF-{\alpha}$). We picked the alveolar macrophage out of mice and cultured it. We analyzed the cytokine gene expression by reverse transcription-PCR. The results obtained were as follows : 1 . Sochungyong-tang showed inhibitory effects on $IL-1{\beta}$ in time and concentration. 2. Sochungyong-tang showed inhibitory effects on IL-6 in time and concentration. 3. Sochungyong-tang showed inhibitory effects on IL-10 in concentration. 4. Sochungyong-tang showed inhibitory effects on iNOS. 5. Sochungyong-tang showed inhibitory effects on $TGF-{\beta}$ in time and concentration. 6. Sochungyong-tang showed on inhibitory effects on $MIP-1{\alpha},\;MIP-1{\beta},\;MIP-1{\gamma}$, $TCF-{\beta}$, $TNF-{\alpha}$. According to above results, it is supposed that Sochungyong-tang has the inhibitory effects on cytokine gene expression in mouse alveolar macrophage and can be usefully applied for curing inflammatory process of lung disease. Advanced studies are required to investigate the cure mechanism of Sochungyong-tang in the future.

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An Experimental Study on the Antioxidant and Anti-inflammatory Effects of Goryeon-hwa (고련환(苦練丸)의 항산화 및 항염증 효과에 관한 실험 연구)

  • Ye-Seul Yun;Seung-Jeong Yang;Seong-Hee Cho
    • The Journal of Korean Obstetrics and Gynecology
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    • v.37 no.2
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    • pp.1-16
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    • 2024
  • Objectives: The purpose of this study is to evaluate the antioxidant and anti-inflammatory effects of Goryeon-hwan (GRH), which is mentioned in ≪Donguibogam≫ that treats leukorrhea. Methods: In this study, the antioxidant efficacy of GRH was evaluated by measuring the total polyphenol and flavonoid content, DPPH radical scavenging activity, ABTS radical scavenging activity, and ROS production through RAW264.7 cells. The concentration of GRH cytotoxicity was confirmed through the cell viability of RAW264.7 cells, and the production of NO, the production of Cytokine through ELISA assay, and the expression of genes through Real-time PCR were measured to evaluate anti-inflammatory efficacy. Protein phosphorylation and protein expression were measured through Western blot analysis. Results: As a result of the experiment, GRH contained polyphenol and flavonoid, and concentration-dependent increased DPPH radical scavenging activity and ABTS radical scavenging activity and decreased ROS production. The anti-inflammatory efficacy measurement results showed a significant decrease in NO and Cytokine production in the GRH administration group compared to the control group. In terms of gene expression and protein expression, there was a significant decrease in iNOS, COX-2, IL-1β, IL-6, and TNF-α depending on the concentration, and a significant increase in HO-1 and NQO1. Protein phosphorylation measurements showed a concentration-dependent significant decrease in the GRH group at ERK and p38. Conclusions: As a result, the study experimentally confirmed the antioxidant and anti-inflammatory effects of GRH, suggesting that it may be used as a treatment for various gynecological inflammatory diseases including vaginitis.

Regulation of Actin Gene Expression During the Differentiation of Naegleria gruberi

  • Kim, Misook;Lee, Joo-Hun
    • Journal of Microbiology
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    • v.39 no.1
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    • pp.42-48
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    • 2001
  • The regulation of actin gene expression during the differentiation of Naegleria gruberi was examined. Actin mRNA concentration was maximal in amoebae and decreased rapidly after the initiation of differentiation. At 20 min after initiation, the concentration of actin mRNA decreased to 55% of the maximal value. The actin mRNA concentration decreased to the minimum at 80 min (15% of the maximum), and then began to increase slightly at the end of differentiation. This decrease of actin mRNA concentration was regulated by the repression of actin gene transcription based on nuclear run-on transcription experiments. The rates of transcription of actin gene in nuclei prepared at 40 and 80 min after the initiation of differentiation were 50 and 28% of that of nuclei prepared at the beginning of differentiation, respectively. The addition of cycloheximide at the initiation of differentiation inhibited both the rapid decrease in the concentration of actin mRNA and the repression of actin gene transcription. These results suggest that the rapid decrease in the concentration of actin mRNA during the differentiation of N. gruberi is accomplished by the repression of actin gene transcription and this transcriptional regulation requires continuous protein synthesis during the differentiation.

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Antioxidant Effect of Nelumbo nucifera G. Leaf Extract and Inhibition of MITF, TRP-1, TRP-2, and Tyrosinase Expression in a B16F10 Melanoma Cell Line (연잎 추출물의 항산화 활성 및 멜라노마 세포(B16F10)에서 MITF, TRP-1, TRP-2, tyrosinase의 발현 저해 효과)

  • Yoo, Dan-Hee;Joo, Da-Hye;Lee, Soo-Yeon;Lee, Jin-Young
    • Journal of Life Science
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    • v.25 no.10
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    • pp.1115-1123
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    • 2015
  • The purpose of this study was to investigate the potential of Nelumbo nucifera G. leaf (NNL) extract as a cosmetic additive. The electron-donating ability of the NNL extract at a concentration of 1,000 μg/ml was 67.83%. In xanthine oxidase, the inhibition effect of the NNL extract was 92.7% at the same concentration. For whitening effects, tyrosinase inhibition effect of NNL extract was 42.7% at a 1,000 μg/ml concentration. The cell toxicity of the NNL extract was examined in melanoma cells (B16F10) using a 3-[4, 5–dimethyl–thiazol–2–yl]-2, 5-diphenyl-tetrazoliumbromide (MTT) assay. The cell toxicity assay revealed that the NNL extract had a toxicity of 81.61% at a concentration of 1,000 μg/ml The microphthalmia-associated transcription factor (MITF), tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2), and tyrosinase protein expression inhibitory effect by Western blot of NNL extract were measured by a Western blot at concentrations of 25, 50, and 100 μg/ml. At a 100 μg/ml concentration of the NNL extract, the expression of the MITF, TRP-1, TRP-2, and tyrosinase protein was decreased by 69.59%, 27.74%, 67.33%, and 67.78% respectively. The MITF, TRP-1, TRP-2 and tyrosinase mRNA expression inhibitory effect were measured by reverse transcription- polymerase chain reaction (PCR) at concentrations of 25, 50, and 100 μg/ml. GAPDH was used as a positive control. At a concentration of 100 μg/ml of the NNL extract, the expression of MITF, TRP-1, TRP-2, and tyrosinase mRNA was decreased by 67.51%, 71.36%, 85.74%, and 83.64%, respectively. These findings suggest that the NNL extract has antioxidant and whitening effects and that it has great potential as a cosmetic ingredient.

Effects of Korean Red Ginseng extract on tissue plasminogen activator and plasminogen activator inhibitor-1 expression in cultured rat primary astrocytes

  • Ko, Hyun Myung;Joo, So Hyun;Kim, Pitna;Park, Jin Hee;Kim, Hee Jin;Bahn, Geon Ho;Kim, Hahn Young;Lee, Jongmin;Han, Seol-Heui;Shin, Chan Young;Park, Seung Hwa
    • Journal of Ginseng Research
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    • v.37 no.4
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    • pp.401-412
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    • 2013
  • Korean Red Ginseng (KRG) is an oriental herbal preparation obtained from Panax ginseng Meyer (Araliaceae). To expand our understanding of the action of KRG on central nervous system (CNS) function, we examined the effects of KRG on tissue plasminogen activator (tPA)/plasminogen activator inhibitor-1 (PAI-1) expression in rat primary astrocytes. KRG extract was treated in cultured rat primary astrocytes and neuron in a concentration range of 0.1 to 1.0 mg/mL and the expression of functional tPA/PAI-1 was examined by casein zymography, Western blot and reverse transcription-polymerase chain reaction. KRG extracts increased PAI-1 expression in rat primary astrocytes in a concentration dependent manner (0.1 to 1.0 mg/mL) without affecting the expression of tPA itself. Treatment of 1.0 mg/mL KRG increased PAI-1 protein expression in rat primary astrocytes to $319.3{\pm}65.9%$ as compared with control. The increased PAI-1 expression mediated the overall decrease in tPA activity in rat primary astrocytes. Due to the lack of PAI-1 expression in neuron, KRG did not affect tPA activity in neuron. KRG treatment induced a concentration dependent activation of PI3K, p38, ERK1/2, and JNK in rat primary astrocytes and treatment of PI3K or MAPK inhibitors such as LY294002, U0126, SB203580, and SP600125 (10 ${\mu}M$ each), significantly inhibited 1.0 mg/mL KRG-induced expression of PAI-1 and down-regulation of tPA activity in rat primary astrocytes. Furthermore, compound K but not other ginsenosides such as Rb1 and Rg1 induced PAI-1 expression. KRG-induced up-regulation of PAI-1 in astrocytes may play important role in the regulation of overall tPA activity in brain, which might underlie some of the beneficial effects of KRG on CNS such as neuroprotection in ischemia and brain damaging condition as well as prevention or recovery from addiction.

Resveratrol inhibits the protein expression of transcription factors related adipocyte differentiation and the activity of matrix metalloproteinase in mouse fibroblast 3T3-L1 preadipocytes

  • Kang, Nam E;Ha, Ae Wha;Kim, Ji Young;Kim, Woo Kyoung
    • Nutrition Research and Practice
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    • v.6 no.6
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    • pp.499-504
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    • 2012
  • This study attempted to investigate the effects of resveratrol on the differentiation of adipocytes. After cells were treated with various concentrations of resveratrol (0, 10, 20, and 40 ${\mu}mol/L$), adipocyte proliferation, the protein expression of transcription factors, and MMPs' activities were determined. Cell proliferation was inhibited more within 4 days of incubation (P<0.05), and lipid accumulation in adipocyte was significantly inhibited by 93.8%, 92.4% and 91.5%, respectively, after two days of 10, 20, and 40 ${\mu}mol/L$ resveratrol treatment (P<0.05). Six days of incubation with the three resveratrol concentrations caused a significantly decreases of 63%, 59.9%, and 25.1% GPDH activity as a dose-dependent response. The triglyceride concentration also decreased significantly with the increase of resveratrol concentration (P<0.05). The protein expression of CCAAT/enhancer-binding protein (C/$EBP{\beta}$) was decreased significantly by 56% and 30% while $PPAR{\gamma}$ was significantly reduced by 57% and 15% with resveratrol treatments of 20 and 40 ${\mu}mol/L$, respectively (P<0.05). The protein expression of C/$EBP{\alpha}$ was decreased by 83%, 74%, and 38% to increased dosage levels, with significance determined for this decrease from 20 ${\mu}mol/L$ of resveratrol. The protein expression of fatty acid binding protein (FABP4) was decreased significantly by 88%, 72%, and 46% with the increase of resveratrol concentration. The activity of MMP-2 was decreased significantly by 84%, 70%, and 63% while MMP-9 activity was decreased significantly by 74%, 62%, and 39% with the increased resveratrol concentrations of 10, 20, and 40 ${\mu}mol/L$, respectively (P<0.05).

Whitening effects of fermented Trigonotis radicans var. sericea with Lactobacillus brevis in α-MSH-stimulated B16F10 melanoma cells

  • Da-Eun Jeong;Byung-Oh Kim;Young-Je Cho
    • Food Science and Preservation
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    • v.31 no.2
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    • pp.227-234
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    • 2024
  • This study was designed to compare the whitening effects of 60% ethanol extracts of Trigonotis radicans var. sericea (TR) and Lactobacillus brevis-fermented T. radicans var. sericea (FTR). Measurement of cytotoxicity in B16-F10 melanoma cells to confirm the whitening effect, FTR showed higher cell viability than TR. FTR showed inhibitory activity on melanin contents similar to the normal group at concentrations of 50 and 100 ㎍/mL. MITF expression was used to confirm the effect on melanogenesis-related protein expression. TR and FTR showed significant concentration-dependent decrease, and FTR showed lower expressions than the normal group at concentrations of 25, 50, and 100 ㎍/mL. Additionally, the mRNA expression of melanogenesis-related genes (MC1R, Rab27a, TGF-β1 and Myo5a) were measured by RT-qPCR to confirm the whitening effect. In MC1R expression at a concentration of 100 ㎍/mL in FTR showed effective inhibitory activities, and in TGF-β1 expression, TR and FTR both showed effective activities compared to normal groups even at low concentrations. Results of myo5a and Rab27a, a similar pattern was shown, and FTR showed effective inhibitory activities at a concentration of 100 ㎍/mL. As a result, FTR had higher whitening effects through bioconversion and is expected to be a good material for whitening functional cosmetics.