• Title/Summary/Keyword: Western Blot

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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.

The Effect of Jeju Wild Ginseng Extracts on Skin Barrier via Serine-Palmitoyltransferase (제주산양산삼이 세린-팔미토일 전이효소(Serine-Palmitoyltransferase)를 통해 피부 장벽에 미치는 효과에 대한 연구)

  • Kim, Hyo Min;Lee, Jung No;Kim, Jae Moon;Kim, Sung Kyu;Park, Sung-Min
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.2
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    • pp.119-126
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    • 2016
  • Skin is the largest organ that protects the body from the external environmental factors such as smog, cigarette smoke, UV. Protective skin barrier is composed with keratinizational keratinocytes and intercellular lipids such as ceramides, cholesterols and fatty acids combined by the lamellar liquid crystal structure. In this research, we confirmed that the Jeju wild ginseng (JWG) extracts dose-dependently increased the expression of serine-palmitoyltransferase (SPT) protein which is associated with ceramide biosynthesis. In addition, emulsion containing 5% JWG extract was applied on skin of human volunteers for 2 weeks and then significantly reduced transepidermal water loss (TEWL) compared to that of control group. As a results, JWG extract increased the biosynthesis of ceramides that is the key components of the skin lipid through enhancing expression of SPT. In addition, JWG extract reduced TEWL resulting in improvement of skin barrier function. In this context, we suggest that JWG extract could be used as a skin barrier enhancer and moisturing agents in cosmetic fileds.

Effects of Boheotang-gagam on Milk Production and Factors Related Lactation in Postpartum Mice (보허탕가감 투여가 산후 생쥐의 유즙분비량 및 유즙분비 관련 인자에 미치는 영향)

  • Lee, Ah-Yeong;Lee, Eun-Hee;Im, Ji-Yeong;Kim, Hong-Jun;Lee, Chang-Hyun
    • The Journal of Korean Obstetrics and Gynecology
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    • v.29 no.1
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    • pp.35-52
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    • 2016
  • Objectives :The purpose of this study was to investigate the effect on factors related the expression of aquaporins (AQP) and milk production after administration of Boheotang-gagam in lactating mice. Methods: The SKH-1 mice were randomly allocated to the control group which was administered with distilled water for two weeks after the parturition and the experimental groups such as, lactating+400G group (L400G) which was administered with Boheotang-gagam 400 mg/day, lactating+600G group (L600G) which was administered with 600 mg/day for two weeks after the parturition, and 400G+lactating+400G group (400G-L400G) which was administered with 400 mg/day for 3 weeks starting one week prior to parturition for experiment (n=6 per group). Results: 1. With regard to the immunohistochemical staining reaction for AQP1, AQP3, and AQP5, stronger immune response was also showed in mammary gland in all experimental groups as compared to the control group. AQP1 showed stronger immune response in the capillaries and venules which were located around the interlobular duct, while stronger immune response of AQP3 and AQP5 showed in the secretory alveolar epithelia and intralobular and interlobular ductal epithelial cells. 2. In the western blot, L400G group showed the most increased expression followed by L600G and then 400G-L400G group in AQP1. In AQP3, the order of expression density was observed as L600G, 400G-L400G and L400G group. Lastly, in AQP5, L400G group presented the most increased expression followed by L600G and 400G-L400G group. Conclusions: Boheotang-gagam would have the effect of increasing the lactation of mice after the birth by increasing the prolactin level and adjusting the expression of AQPs and prolactin receptor in the mammary glands.

The Effect of Hwangryunhaedok-tang on the c-fos Level in Mice Exposed to Heat Stress (황련해독탕(黃連解毒湯)이 외열 스트레스(화사(火邪))에 의한 동물모델에서 c-fos 발현에 미치는 영향 연구)

  • Hwang, Deok-Sang;Kim, Hyo-Geun;Kim, Tae-Mi;Lee, Tae-Hee;Oh, Myung-Sook
    • The Journal of Korean Obstetrics and Gynecology
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    • v.27 no.3
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    • pp.1-11
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    • 2014
  • Objectives: Hwangryunhaedok-tang (HH) is a representative heat clearing multiherb prescription. We evaluated the protective effect of HH against heat stress exposure in mice. Methods: Six weeks old ICR mice were used for this study. After $43^{\circ}C$ heat stress for 15 minutes, we evaluated the changes of motor activity and c-fos expression level to determine the proper heat stress and evaluation time. The subjects were divided into 4 groups (1. control group, 2. heat stress group with normal saline, 3. heat stress group with administration of 100 mg/kg, and 4. heat stress group with administration of 1000 mg/kg). After oral administration of HH once a day for 3 days, 2, 3, 4 groups were exposed to $43^{\circ}C$ heat stress for 15 minutes. Then, we evaluated the motor activity for 120 minutes and analyzed the c-fos expression using western blot. We investigated the effect of HH and its ingredients on c-fos expressions after heat stress. The mice were administrated HH, Scutellariae Radix, Coptidis Rhizoma, Phellodendri Cortex, and Gardeniae Fructus for 3 days. After one hour of last treatment, the mice were exposed to heat stress at $43^{\circ}C$ for 15 min. After two hours, the hypothalamus was dissociated and lysed to measure c-fos expression. Results: After oral administration of HH for 3 days, motor activity was recovered insignificantly. 100 mg/kg HH treatment reduced the c-fos expression after heat stress but insignificantly. Among the ingredients of HH, Coptidis Rhizoma and Scutellariae Radix treatment groups significantly reduced the c-fos after stress. Conclusions: These results show that Hwangryunhaedok-tang may be effective to reduce the heat stress response.

Effect of Astragali Radix Pharmacocupuncture for Wound Healing of Full-Thickness Skin Wound in Mouse (황기약침이 전층피부손상 마우스의 치유에 미치는 영향)

  • Hong, Chang-Ho;Kang, Kyung-Hwa;Yoon, Hwa-Jung;Yoon, Hyoun-Min;Song, Choon-Ho;Kim, Cheol-Hong
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.31 no.1
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    • pp.22-31
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    • 2018
  • Objectives : In this study, we investigated whether Astragali Radix Pharmacocupuncture (ARP) has an effective on the full thickness defect wound healing process of mouse. Methods : A total of 50 mice (ICR mouse, 7 week-old male) were divided into control group and ARP group. A single full thickness skin defect was made on the dorsal side of the each mouse using an 8mm diameter biopsy punch. Control group were treated with 0.2㎖ saline and ARP group were treat with 0.2ml ARP at 8 points around the wound every three days total 4 times during the experimental period. The change in wound size, contraction rate, healing rate, and epithelization rate was measured by digital images taken on days 3, 6, 9, and 13, and evaluated using a digital image analysis program. Tissues were collected for histological analysis, RT-PCR, and Western blot on days 3, 6, 9, and 15. Results : The results are as follows. ARP group accelerated the rate of wound contraction, wound healing and epithelization compared to the control group. ARP group showed the decrease of inflammatory cells in early inflammatory phase compared to the control group. ARP group upregulated PECAM-1 mRNA and protein expression compared to the control group. ARP group inhibited the scar width and area compared to the control group. Conclusions: ARP showed positive effects on wound healing through the inhibition of inflammatory reaction and increase of PECAM-1 expression related to the wound healing process.

Antioxidant activities and anti-inflammatory effects of fresh and air-dried Abeliophyllum distichum Nakai leaves (건조방법에 따른 미선나무 잎의 항산화 및 항염증 효과)

  • Chang, Seong Jun;Jeon, Nam Bae;Park, Joo Won;Jang, Tae Won;Jeong, Jin Boo;Park, Jae Ho
    • Food Science and Preservation
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    • v.25 no.1
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    • pp.27-35
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    • 2018
  • In this study, we evaluated the antioxidant activity and anti-inflammatory effects of Abeliophyllum distichum (A. distichum) leaves that were prepared via air-drying. Fresh and air-dried A. distichum leaves were examined via 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical, and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging assay and measurements of the reducing power. The suppression effects on inflammation of the leaves were analyzed by a western blot and RT-PCR on LPS-induced RAW 264.7 cells. As a result, the antioxidant activity of the fresh leaves was found to be more effective than that of the air-dried leaves. Also, the fresh leaves were more effective in suppressing the protein and mRNA levels of iNOS and COX-2 than the air-dried leaves, thereby indicating the better anti-inflammatory effects. In addition, the contents of phenolic compounds and acteoside were analyzed by high-performance liquid chromatography (HPLC). The results showed that the acteoside content decreased with the use of the air-drying method, while there was no change in the content of phenolic compounds. Therefore, this study indicated that fresh A. distichum leaves potential antioxidant and suppression activities of various factors that are involved in the production of NO, which were found to be better than those of air-dried A. distichum leaves. These biological activities were also found to be independent of the content of phonolic compounds and were assumed to be directly or indirectly related to the content of acteoside.

Dietary glucosinolates inhibit splenic inflammation in high fat/cholesterol diet-fed C57BL/6 mice

  • Gu, HyunJi;Gwon, Min-Hee;Kim, Sang-Min;Yun, Jung-Mi
    • Nutrition Research and Practice
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    • v.15 no.6
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    • pp.798-806
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    • 2021
  • BACKGROUND/OBJECTIVES: Obesity is associated with chronic inflammation. The spleen is the largest organ of the lymphatic system and has an important role in immunity. Obesity-induced inflammatory responses are triggered by Toll-like receptor (TLR)-myeloid differentiation primary response 88 (MyD88) pathway signaling. Phenethyl isothiocyanate (PEITC) and 3,3'-diindolylmethane (DIM), major dietary glucosinolates present in cruciferous vegetables, have been reported to produce anti-inflammatory effects on various diseases. However, the effects of PEITC and DIM on the obesity-induced inflammatory response in the spleen are unclear. The purpose of this study was to examine the anti-inflammatory effects of PEITC and DIM on the spleen and their mechanism in high fat/cholesterol diet (HFCD)-fed C57BL/6 mice. MATERIALS/METHODS: We established an animal model of HFCD-induced obesity using C57BL/6 mice. The mice were divided into six groups: normal diet with AIN-93G diet (CON), high fat diet (60% calories from fat) with 1% cholesterol (HFCD), HFCD with PEITC 30 mg/kg/day or 75 mg/kg/day (HFCD+P30, HFCD+P75), and HFCD with DIM 1.5 mg/kg/day or 7.5 mg/kg/day (HFCD+D1.5, HFCD+D7.5). Enzyme-linked immunosorbent assay was used to evaluate pro-inflammatory cytokine secretion. Western blot and quantitative polymerase chain reaction were used to analyze protein and mRNA levels of nuclear factor kappa B (NF-κB) p65, interleukin 6 (IL-6), cyclooxygenase 2 (COX-2), TLR2, TLR4, and MyD88 in spleen tissue. RESULTS: Serum IL-6 levels were significantly higher in the HFCD group than in groups fed a HFCD with PEITC or DIM. Levels of NF-κB p65 protein and TLR2/4, MyD88, NF-κB p65, IL-6, and COX-2 mRNA were significantly higher in the HFCD group than in the CON group and were reduced by the PEITC and DIM supplements. CONCLUSIONS: PEITC- and DIM-supplemented diets improved splenic inflammation by modulating the TLR2/4-MyD88 pathway in HFCD-fed mice. We suggest that dietary glucosinolates may at least partially improve obesity-induced inflammation of the spleen.

Effect of Paeoniae Radix Alba on a thioacetamide induced liver fibrosis mice model (Thioacetamide로 유발된 간섬유증 동물 모델에서 백작약이 미치는 효능)

  • Lee, Se Hui;Lee, Jin A;Shin, Mi-Rae;Seo, Bu-Il;Roh, Seong-Soo
    • Korean Journal of Food Science and Technology
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    • v.53 no.5
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    • pp.544-552
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    • 2021
  • This study investigated the anti-fibrotic and antioxidant effects of Paeonia Radix Alba water extract (PR) on thioacetamide (TAA)-induced liver fibrosis in a mouse model and its underlying mechanisms. Liver fibrosis was induced by intraperitoneal injection of TAA (three times a week) for 8 weeks. Furthermore, silymarin (50 mg/kg body weight) and PR (200 mg/kg body weight) were administered for 8 weeks. PR treatment downregulated aspartate aminotransferase (AST), alanine aminotransferase (ALT), ammonia, and myeloperoxidase levels. Moreover, PR treatment downregulated NOX2 and p47phox and upregulated antioxidant enzymes by activating the Nrf2/Keap1 signaling pathway. Furthermore, PR inhibited the factors associated with fibrosis, such as α-SMA and collagen I. AMPK/SIRT1 was upregulated by PR treatment. Overall, these results suggest that PR attenuates liver fibrosis by regulating the Nrf2/Keap1 and AMPK/SIRT1/NF-κB signaling pathways through the inhibition of oxidative stress. Hence, PR has potential as a remedy for preventing and treating liver fibrosis.

Psoraleae Semen Ethanol Extract Inhibits RANKL-Induced Osteoclast Differentiation and Osteoclast Specific Genes Expression (보골지 추출물이 파골세포 분화 및 골흡수 관련 유전자 발현에 미치는 영향)

  • Ryu, Gwang-hyun;Kim, Eom Ji;Kim, Minsun;Kim, Jae-Hyun;Lee, Yujin;Jin, Dae-hwan;Sohn, Youngjoo;Jung, Hyuk-Sang
    • Korean Journal of Acupuncture
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    • v.38 no.3
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    • pp.140-150
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    • 2021
  • Objectives : The increase of osteoclasts could cause osteoporosis and bone-related diseases. Also, the inhibition of osteoclast differentiation is important in treating bone-related diseases. Traditionally, Psoraleae Semen has been used for geriatric diseases, aging and musculoskeletal diseases. The purpose of this study is to investigate the effect of Psoraleae Semen ethanol extract (PS) on osteoclast differentiation and its function. Methods : To confirm the effect of PS on osteoclastogenesis and bone resorption activity, various levels of concentrations of PS (5, 10, 20 and 40 ㎍/ml) were tested on RAW 264.7 cells cultured with RANKL. We measured tartarate-resistant acid phosphatase (TRAP)-positive cells, TRAP activity, pit formation and F-actin ring formation. The expressions of nuclear factor of activated T-cells (NFATc1) and c-Fos were confirmed through western blot and reverse transcription- polymerase chain reaction (RT-PCR). Also, the expression of bone resorption and fusion-related genes in osteoclast was confirmed by RT-PCR. Results : PS decreased the number of TRAP-positive cells and the TRAP activity. In addition, PS significantly inhibited the formation of pit and F-actin ring. Furthermore, PS decreased the expression of osteoclast related genes. Conclusions : PS inhibits osteoclast differentiation and bone resorption ability through inhibition of the expression of osteoclast-related genes. This indicates that PS may be a potential therapeutic agent to osteoporosis by suppressing osteoclastogenesis.

A Study on the Anti-Pollution Effect of Kombucha Fraction through AhR Expression (AhR 발현을 통한 콤부차 분획물의 안티폴루션 효과 연구)

  • Kang, Eun-Bin;Hyun, Jin-A;Kwon, Hyun-Ji;Beom, Seok-Hyun;Han, Dong-Geun;Park, Ha-Eun;Kim, Hyun-Jeong;Kwak, Ki-Sung;An, Bong-Jeon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.47 no.3
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    • pp.213-218
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    • 2021
  • In this study, Kombucha, a fermented drink by adding beneficial bacteria to green tea or black tea with sugar, was classified using column chromatography, and the presence or absence of quercetin glycoside, a type of flavonoids, was confirmed through TLC. In addition, the anti-pollution effect of protecting and improving the skin from pollution was confirmed using the fraction. Keratinocytes were treated with K-QG to confirm the cell viability, showing a survival rate of 90% or more up to a concentration of 100 ㎍/mL, and the cell viability by benzo[e]pyrene and fine dust stimulation to see the anti-pollution effect was measured. At 100 ㎍/mL, the improvement rates were about 68.79% and 50.68%, respectively. In addition, as a result of confirming the expression of AhR activated by benzo[a]pyrene through Western blot, it showed an inhibition rate of about 31.08% at 100 ㎍/mL compared to the control group. Through the results of this study, K-QG is thought to be valuable as an anti-pollution functional material, protecting and improving skin irritated from benzopyrene and fine dust.