• Title/Summary/Keyword: Western Blot

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Study on Applicability of Pinus koraiensis Siebold et Zucc Leaf Extract as a Cosmetic Ingredient (잣나무(Pinus koraiensis Siebold et Zucc)잎 추출물의 화장품 소재로서의 활용 가능성에 관한 연구)

  • Jeon, Myung-Ok;Moon, Ji-sun
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.3
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    • pp.602-612
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    • 2017
  • This study was performed to verify various effects of 70% ethanol extract and distilled water extract of pine leaf such as antioxidant, anti-inflammatory, skin-lightening effects analysis in cells. The result of antioxidant experiment showed that polyphenol content increased concentration dependently in polyphenol and flavonoid effects. The total content of polyphenols and flavonoids was much higher in ethanol extract than water extract. In all the cells of B16F10, RAW264.7 cytotoxicity at each concentration level didn't appear. The result of measuring NO production inhibition showed that it inhibited NO production led to LPS effectively, so anti-inflammatory activity was confirmed. The result of measuring melanin biosynthesis inhibition showed that there was considerably reduction effect, but it performed western blot, and as a result of MITF, Tyrosinase protein revelation, the inhibition of MITF, Tyrosinase revelation was confirmed concentration dependently, Therefore, in this study pine leaf extract was expected to be used as a cosmetic ingredient.

The inhibition of inflammatory molecule expression on 3T3-L1 adipocytes by berberine is not mediated by leptin signaling

  • Choi, Bong-Hyuk;Kim, Yu-Hee;Ahn, In-Sook;Ha, Jung-Heun;Byun, Jae-Min;Do, Myoung-Sool
    • Nutrition Research and Practice
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    • v.3 no.2
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    • pp.84-88
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    • 2009
  • In our previous study, we have shown that berberine has both anti-adipogenic and anti-inflammatory effects on 3T3-L1 adipocytes, and the anti-adipogenic effect is due to the down-regulation of adipogenic enzymes and transcription factors. Here we focused more on anti-inflammatory effect of berberine using real time RT-PCR and found it changes expressions of adipokines. We hypothesized that anti-adipogenicity of berberine mediates anti-inflammtory effect and explored leptin as a candidate mediator of this signaling. We studied this hypothesis by western blot analysis, but our results showed that berberine has no effect on the phosphorylations of STAT-3 and ERK which have important roles on leptin signaling. These results led us to conclude that the anti-inflammatory effect of berberine is not mediated by the inhibition of leptin signal transduction. Moreover, we have found that berberine down-regulates NF-${\kappa}B$ signaling, one of the inflammation-related signaling pathway, through western blot analysis. Taken together, the anti-inflammatory effect of berberine is not mediated by leptin, and berberine induces anti-inflammatory effect independent of leptin signaling.

Anti-inflammatory Effects of Korean red ginseng Extract in formalin-induced Orofacial Pain in Rats (포르말린으로 유도된 안면통증에 대한 홍삼추출물의 항염증 효과)

  • Jin, Byung-Moon;Lee, Min-Kyung;Lee, Jun-Seon;Hyun, Kyung-Yae
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.15 no.9
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    • pp.5708-5715
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    • 2014
  • The effects of korean red ginseng (KRG) extracts on orifacial pain control in terms of the systemic inflammatory response and pharmacological effects as health supplements were investigated. The experimental group were divided into three groups, the control group (n=6), formalin (5%, $50{\mu}{\ell}$) injection group (n=6), and formalin (5%, $50{\mu}{\ell}$) injection added KRG administrated group (4.5 ml/kg, n=6). The KRG administrated group prior to the formalin injection significantly attenuated the behavioral response compared to that of the control group. Pain reduction was suppressed mainly from 15 min to 30 min. The KRG administrated rats showed significantly reduced p38 MAPK, iNOS and Nrf2 expression in the brain and medulla oblongata according to Western blot analysis. These findings suggest that KRE may have a useful effect on orificial pain control functions by preventing the p38 MAPK pathway.

The Study on Apoptosis and Expression of Fas, Fas-ligand, Bax, and Bcl-2 in Human Fragmented Embryos (분절화된 인간 배아에서 세포자연사와 Fas, Fas-ligand, Bax, Bcl-2 발현에 관한 연구)

  • Kim, Jong-Sik;Kim, Myoung-Shin;Yang, Hyun-Won;Yu, Chai-Hyeock;Yoon, Yong-Dal;Bae, In-Ha;Jung, Byeong-Jun;Song, Hyun-Jin
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.3
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    • pp.167-178
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    • 2002
  • Objective : The present study was performed to investigate whether apoptosis occur in human embryos by annexin staining and detect the expression of Fas, Fas-ligand (FasL), Bax, and Bcl-2 in human fragmented embryos derived from IVF-ET by immunofluorescence and Western blot analysis. Materials and Methods: Using annexin staining, immunofluorescence and Western blot analysis on normal and fragmented embryos, we were able to detect apoptotsis and apoptotic gene products in fragmented embryos. Result: Phosphatidylserine (PS) translocation, the marker for apoptosis, were detected frequently in fragmented embryos. Bcl-2 and Bax protein were detected in both fragmented and non-fragmented embryos. When fragmented embryos compared to normal embryos, immunofluorescent intensity of Bcl-2 tended to be lower in fragmented embryos. Bax gene expression increased in the fragmented embryos compared to the normal embryos. This result supports a model in which the molar ratio of Bcl-2 to Bax determines whether apoptosis induced or inhibited in human embryo. Fas was highly expressed in human preimplantation embryos but not FasL. It suggests that embryo may undergo apoptosis by binding with FasL produced by follicular or immune cells. Conclusion: The over expression of Bax and Fas will trigger apoptosis to lead embryo fragmentation and change embryo to be nonviable.

Screening and Development of DNA Aptamers Specific to Several Oral Pathogens

  • Park, Jung-Pyo;Shin, Hye Joo;Park, Suk-Gyun;Oh, Hee-Kyun;Choi, Choong-Ho;Park, Hong-Ju;Kook, Min-Suk;Ohk, Seung-Ho
    • Journal of Microbiology and Biotechnology
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    • v.25 no.3
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    • pp.393-398
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    • 2015
  • Aptamers are composed of single-stranded oilgonucleotides that can selectively bind desired molecules. It has been reported that RNA or DNA could act as not only a genetic messenger but also a catalyst in metabolic pathways. RNA aptamers (average sizes 40-50 bp) are smaller than antibodies and have strong binding capacities to target molecules, similar to antigenantibody interactions. Once an aptamer was selected, it can be readily produced in large quantities at low cost. The objectives of this study are to screen and develop aptamers specific to oral pathogens such as Porphyromonas gingivalis, Treponema denticola, and Streptococcus mutans. The bacterial cell pellet was fixed with formaldehyde as a target molecule for the screening of aptamers. The SELEX method was used for the screening of aptamers and a modified western blot analysis was used to verify their specificities. Through SELEX, 40 kinds of aptamers were selected and the specificity of the aptamers to the bacterial cells was confirmed by modified western blot analysis. Through the SELEX method, 40 aptamers that specifically bind to oral pathogens were screened and isolated. The aptamers showed possibility as effective candidates for the detection agents of oral infections.

Protective Effects of Celastrol, the Triterpenoid Component of Celastrus Orbiculatus, on Dopaminergic Neuronal Cells in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-lesioned Parkinson's Disease Rats (노박덩굴에 함유된 celastrol 성분의 파킨슨병을 유발시킨 쥐에서의 도파민 신경세포 보호효과)

  • Lee, Kap-Duk;Kim, Kwang-Jin;Park, Yong-Ki
    • The Journal of Korean Medicine
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    • v.29 no.4
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    • pp.94-103
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    • 2008
  • Objectives: The aim of this study was to determine whether celastrol, the triterpenoid component of Celastrus orbiculatus, offers neuroprotection against Parkinson's disease (PD) in mice administered 1-methyl-4-phenyl-1,2,3,6- tetrahydropyridine(MPTP). Methods: We examined how celastrol affected MPTP-induced neuronal loss of tyrosine hydroxylase (TH)-positive dopaminergic neurons in substantia nigra pars compacta (SNpc) in the midbrain of mice. C57BL/6J mice were divided into four groups: (1) saline-saline, (2) saline-celastrol, (3) MPTP-saline, and (4) MPTP-celastrol. The mice were injected intraperitoneally (i.p.) with four administrations of MPTP (18mg/kg) at 2 h intervals and then i.p. administered celastrol (3mg/kg) two times at 12 h after last celastrol administration. Expression of TH on the SNpc of brain tissues were analyzed at 7 days after the treatments by immunohistochemistry and Western blot. Results: Immunohistochemical analysis using TH antibody showed that celastrol provided significantly protective effects against MPTP-induced loss of TH-positive dopaminergic neurons in the SNpc region of the midbrain of mice. Our Western blot study also showed that celastrol significantly inhibits the MPTP-induced neuronal damage via the up-regulation of TH protein levels in MPTP mice. Conclusions: The present results suggest that it may be possible to use celastrol for the prevention of nigral degenerative disorders including PD, caused by exposure to toxic substances.

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Gecko Proteins Exert Anti-Tumor Effect against Cervical Cancer Cells Via PI3-Kinase/Akt Pathway

  • Jeong, Ae-Jin;Chung, Chung-Nam;Kim, Hye-Jin;Bae, Kil-Soo;Choi, Song;Jun, Woo-Jin;Shim, Sang-In;Kang, Tae-Hong;Leem, Sun-Hee;Chung, Jin-Woong
    • The Korean Journal of Physiology and Pharmacology
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    • v.16 no.5
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    • pp.361-365
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    • 2012
  • Anti-tumor activity of the proteins from Gecko (GP) on cervical cancer cells, and its signaling mechanisms were assessed by viable cell counting, propidium iodide (PI) staining, and Western blot analysis. GP induced the cell death of HeLa cells in a dose-dependent manner while it did not affect the viability of normal cells. Western blot analysis showed that GP decreased the activation of Akt, and co-administration of GP and Akt inhibitors synergistically exerted anti-tumor activities on HeLa cells, suggesting the involvement of PI3-kinase/Akt pathway in GP-induced cell death of the cancer cells. Indeed, the cytotoxic effect of GP against HeLa cells was inhibited by overexpression of constituvely active form of Akt in HeLa cells. The candidates of the functional proteins in GP were analyzed by Mass-spectrum. Taken together, our results suggest that GP elicits anti-tumor activity against HeLa cells by inhibition of PI3-kinase/Akt pathway.

Cisplatin-induced Alterations of $Na^+$-dependent Phosphate Uptake in Renal Epithelial Cells

  • Lee, Sung-Ju;Kwon, Chae-Hwa;Kim, Yong-Keun
    • The Korean Journal of Physiology and Pharmacology
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    • v.11 no.2
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    • pp.71-77
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    • 2007
  • Cisplatin treatment increases the excretion of inorganic phosphate in vivo. However, the mechanism by which cisplatin reduces phosphate uptake through renal proximal tubular cells has not yet been elucidated. We examined the effect of cisplatin on $Na^+$-dependent phosphate uptake in opossum kidney (OK) cells, an established proximal tubular cell line. Cells were exposed to cisplatin for an appropriate time period and phosphate uptake was measured using $[^{32}P]$-phosphate. Changes in the number of phosphate transporter in membranes were evaluated by kinetic analysis, $[^{14}C]$phosphonoformic acid binding, and Western blot analysis. Cisplatin inhibited phosphate uptake in a time- and dose-dependent manner, and also the $Na^+$-dependent uptake without altering $Na^+$-independent uptake. The cisplatin inhibition was not affected by the hydrogen peroxide scavenger catalase, but completely prevented by the hydroxyl radical scavenger dimethylthiourea. Antioxidants were ineffective in preventing the cisplatin-induced inhibition of phosphate uptake. Kinetic analysis indicated that cisplatin decreased Vmax of $Na^+$-dependent phosphate uptake without any change in the Km value. $Na^+$-dependent phosphonoformic acid binding was decreased by cisplatin treatment. Western blot analysis showed that cisplatin caused degradation of $Na^+$-dependent phosphate transporter protein. Taken together, these data suggest that cisplatin inhibits phosphate transport in renal proximal tubular cells through the reduction in the number of functional phosphate transport units. Such effects of cisplatin are mediated by production of hydroxyl radicals.

Eupatilin downregulates phorbol 12-myristate 13-acetate-induced MUC5AC expression via inhibition of p38/ERK/JNK MAPKs signal pathway in human airway epithelial cells

  • Cheon, Yoon-Hee;Kim, Min Seob;Kim, Ju-Young;Kim, Dong Hyun;Han, Seung Yoon;Lee, Jae-Hoon
    • The Korean Journal of Physiology and Pharmacology
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    • v.24 no.2
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    • pp.157-163
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    • 2020
  • Chronic inflammatory airway diseases, such as chronic rhinosinusitis, chronic obstructive pulmonary disease, and asthma, are associated with excessive mucus production. Hence, the regulation of mucus production is important for the treatment of upper and lower airway diseases. Eupatilin is a pharmacologically active ingredient obtained from Artemisia asiatica Nakai (Asteraceae) and exerts potent anti-inflammatory, anti-allergic, and anti-tumor activities. In the present study, we investigated the effect of eupatilin on phorbol 12-myristate 13-acetate (PMA)-induced MUC5AC and MUC5B expression in human airway epithelial cells. We found that eupatilin treatment significantly inhibited PMA-induced mucus secretion in PAS staining. In addition, qRT-PCR results showed that eupatilin dose-dependently decreased the mRNA expression of MUC5AC in human airway epithelial cells. Western blot and immunofluorescence assay also showed that PMA-induced protein expression of MUC5AC was inhibited by eupatilin treatment. Finally, we investigated MAPKs activity after stimulation with PMA using western blot analysis in human airway epithelial cells. The results showed that eupatilin downregulated the levels of phosphorylated p38, ERK, and JNK. In summary, the anti-inflammatory activities of eupatilin, characterized as the suppression of MUC5AC expression and secretion in human airway epithelial cells, were found to be associated with the inhibition of p38/ERK/JNK MAPKs signaling pathway of MUC5AC secretion.

Effect of Red Ginseng Extract on the Inflammatory Response of LPS-Induced RAW 264.7 cell (홍삼추출물이 LPS로 유도된 RAW 264.7 cell의 염증반응에 미치는 효과)

  • Jang, Young-Ah;Kim, Han-Na;Kim, Bo-Ae
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.4
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    • pp.1434-1442
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    • 2019
  • We conducted this study to investigate anti-inflammatory possibilities of applying cosmetic material about extracts from red ginseng. For this we carried out biological active evaluation about anti-inflammatory by using extracts of red ginseng. In order to evaluate the anti-inflammatory effects of the samples in macrophages (RAW 264.7 cells), MTT assay was used to evaluate the toxicity of red ginseng extracts and the inhibitory activity of nitric oxide production and the expression levels of inflammation-related proteins and genes. The inhibitory activity of nitric oxide in the LPS-induced RAW 264.7 cells was 71.2% at 25 ㎍/ml concentration and western blot analysis showed that the expression of iNOS and COX-2 protein decreased in a concentration-dependent manner. These results suggest that extracts from red ginseng may have value as the potential cosmetic materials.