• Title/Summary/Keyword: RAW264.7

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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.

Studies on Efficient Extraction of Limonene from Citron and Immune-modulation Activity for Development of Environmentally Friendly Material (친환경 소재 개발을 위한 유자에서의 효율적 Limonene 추출 및 면역기능 조절활성에 관한 연구)

  • Ahn, Jong-Ho;Lim, Hyun-Hee;Hwang, Seong-Gu;Nam, In-Sik
    • Korean Journal of Organic Agriculture
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    • v.28 no.4
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    • pp.591-604
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    • 2020
  • The objectives of this study were to find out the best condition of extracting methods of limonene from citron and to determine effects of limonene on immune modulation activity by measuring cytokine secretion using RAW 264.7 mouse macrophage cells. When distilled water was used as a solvent instead of organic solvents to extract limonene from citron, addition of refluxing process to simultaneous steam distillation extraction method was found to be much effective in extracting limonene. However, it required longer extraction time than using other organic solvents. Limonene extracts showed increased IL-β and IL-6 but decreased the TNF-α gene expression in limonene concentration dependant manner. However oral administration of limonene extracts to mice did not influence significantly compared to control in in vivo experiment. It might be due to that the mice were kept in well controlled and complete environment. Limonene, a natural material from citron has been approved to have a immune-modulation activity in the present study and have a potential as a feed additive that is environmentally friendly and no harmful. Further study with protected limonene, for example, for the protection of limonene from oxidation or bypass the ruminal degradation in order consequently to increase immune-modulation activity might be useful as a further research.

An Experimental Study on the Anti-inflammatory, Anti-pruritic and Anti-microbial Effects of the Three Herbal Prescription: Eunhwasagan-tang (EST), Sobokchukeo-tang (SCT), Wandae-tang (WDT) (대하(帶下) 처방 3종의 항염, 항소양, 항균 효능에 관한 실험 연구 : 은화사간탕(銀花瀉肝湯), 소복축어탕(少腹逐瘀湯), 완대탕(完帶湯))

  • Lee, Eun-Kyu;Park, Chan-Wook;Kim, Soo-Hyeon;Choi, Yoo-Jin;Park, Kyung-Mi;Yang, Seung-Jeong;Cho, Seong-Hee
    • The Journal of Korean Obstetrics and Gynecology
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    • v.32 no.3
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    • pp.32-56
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    • 2019
  • Objectives: The purpose of this study was to investigate the in vitro anti-inflammatory, anti-pruritic and antimicrobial effects of the three herbal prescription (EST, SCT, WDT), which has been traditionally used for treating leukorrhea induced by various infections in the female genital tract. Methods: In this experiment, the anti-inflammatory effects were evaluated by Nitric oxide (NO), $Interlukine-1{\beta}$ ($IL-1{\beta}$), Interlukine-2 (IL-2), Interlukine-6 (IL-6), Tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), Prostaglandin $E_2$ ($PGE_2$), Leukotriene $B_4$ ($LTB_4$) production amount and Inducible nitric oxide synthase (iNOS), Nuclear factor kappa B ($NF-{\kappa}B$), Cyclooxygenase-2 (COX-2) gene expression levels in RAW264.7 cells. And the anti-pruritic effects were evaluated by Histamine, Acetylcholine (ACh), Acetylcholinesterase (AChE), Substance P production amount in Mast cell/9 (MC/9) and Pheochromocytoma 12 (PC12) cells. The anti-microbial effect was measured by inhibition zone diameter on Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans and Aspergillus niger. Results: As a result of measuring anti-inflammatory efficacy, $IL-1{\beta}$, IL-2, IL-6, $TNF-{\alpha}$, $PGE_2$, and $LTB_4$ production amounts were significantly reduced in the EST, SCT, WDT extraction groups compared with the control group, and significantly decreased the amount of $NF-{\kappa}B$, iNOS, and COX-2 gene expression and the amount of Phospho-Inhibitor kappa B alpha ($p-I{\kappa}B-{\alpha}$)/Inhibitor kappa B alpha ($I{\kappa}B-{\alpha}$) and $NF-{\kappa}B$ p65 protein expression. In addition, As a result of measuring the anti-pruritic effect, the amounts of histamine, ACh and Substance P were significantly decreased, and AChE production was slightly decreased, but it's significance did not appear. Finally the anti-microbial effects of EST, SCT, WDT extraction groups against Pseudomonas aeruginosa, Candida albicans and Aspergillus niger was inhibited, however the growth of Escherichia coli and Staphylococcus aureus was not inhibited. Conclusions: These data suggest that EST, SCT, WDT can be used to treat patients with leukorrhea.

The Content of Amino Acids and Anti-inflammatory Effect of Lentinula edodes Syrup by Different Malt Concentrations (맥아 첨가비율을 달리한 표고당화액의 세포독성, 항염증효과 및 아미노산 함량)

  • Ha, Neul-I;Bak, Ok Ran;Kim, Kyung Je;Jin, Seong Woo;Koh, Young Woo;Im, Seung Bin;Jeong, Hee Gyeong;Seo, Kyoungsun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.104-104
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    • 2019
  • 표고(Lentinula edodes)에 다량 함유되어 있는 아미노산은 면역효과와 밀접한 관련이 있는 것으로 알려져 있다. 표고 특유의 향미에 대한 소비자들의 선호도가 상이하여, 표고를 맥아로 당화하여 소비자들의 섭취 편이성을 향상시키고, 아미노산을 포함한 당화액을 제조하여 면역효과가 증강된 제품을 개발하고자 본 연구를 수행하였다. 또한 버섯류를 비롯한 담자균류에는 다양한 약효성분이 함유되어 있어 민간에서도 자주 사용된 기록이 있다. 최근 고등담자균류인 버섯의 성분 및 유용성에 관심이 크게 증가하고 있는 추세이며, 이에 따라 버섯류에 대한 식품학적 및 약리학적 측면에서 연구가 활발히 진행되고 있다. 표고(L.edodes)와 당화효소로 사용한 맥아의 비율에 따라 A (표고 1 kg : 맥아 50 g), B (표고 1 kg : 맥아 100 g), C (표고 1 kg : 맥아 200 g), D (표고 1 kg : 맥아 300 g), E (표고 1 kg : 맥아 300 g) 5개의 시험구를 설정하였으며, 제조된 표고당화액을 농축 후 동결건조 하여 분말로 분쇄 후 시료로 사용하였다. 표고당화액의 면역효과를 검증하고자, Raw 264.7에 표고당화추출분말 A, B, C, D, E를 각각 10, 50, 100 및 $500{\mu}g/mL$의 농도로 처리하여 각 시료에 대한 세포 독성과 항염증효과를 확인하였다. 세포독성 시험결과, E 시험구의 경우 $100{\mu}g/mL$의 농도에서 75.3%, $500{\mu}g/mL$의 농도에서 66.3%의 세포생존율을 나타내 독성을 보였다. 또한 A, B, D를 각각 $500{\mu}g/mL$의 농도로 처리 했을 때, 94.1%, 83.1%, 80.2%의 NO 생성율을 나타내어 E 시험구를 제외하고는 세포독성이 없는 것으로 확인되었다. 표고당화추출분말의 구성아미노산 함량은 C 시험구에서 19,699.01 mg%로 가장 높게 나타났으며, A 시험구에서 17,231.14 mg%, B 시험구에서 17,152.38 mg%순으로 높게 나타났다. E 시험구에서 13,367.98 mg%로 가장 낮게 나타났다. 모든 표고당화추출분말에서 총 16종의 유리아미노산 이 검출 되었으며, C 시험구에서 2,562.06 mg%로 가장 높게 나타났고, B 시험구에서 2,243.16 mg%, D 시험구에서 1,895.29 mg%, A 시험구에서 1,672.82 mg%순으로 높게 나타났다.

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Evaluation of in vitro anti-oxidant and anti-inflammatory activities of Korean and Chinese Lonicera caerulea

  • Lee, You-Suk;Cho, Il Je;Kim, Joo Wan;Lee, Sun-Kyoung;Ku, Sae Kwang;Lee, Hae-Jeung
    • Nutrition Research and Practice
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    • v.12 no.6
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    • pp.486-493
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    • 2018
  • BACKGROUND/OBJECTIVE: The honeysuckle berry (HB) contains ascorbic acid and phenolic components, especially anthocyanins, flavonoids, and low-molecular-weight phenolic acids. In order to examine the potential of HB as a hepatoprotective medicinal food, we evaluated the in vitro anti-oxidant and anti-inflammatory activities of Korean HB (HBK) and Chinese HB (HBC). MATERIALS/METHODS: Antioxidant and anti-inflammatory effects of the extracts were examined in HepG2 and RAW 264.7 cells, respectively. The anti-oxidant capacity was determined by DPPH, SOD, CAT, and ARE luciferase activities. The production of nitric oxide (NO) as an inflammatory marker was also evaluated. The Nrf2-mediated mRNA levels of heme oxygenase-1 (HO-1), NAD(P)H dehydrogenase [quinone] 1 (Nqo1), and glutamate-cysteine ligase catalytic subunit (Gclc) were measured. The concentrations of HB extracts used were 3, 10, 30, 100, and $300{\mu}g/mL$. RESULTS: The radical scavenging activity of all HB extracts increased in a concentration-dependent manner (P < 0.01 or P < 0.05). SOD (P < 0.05) and CAT (P < 0.01) activities were increased by treatment with $300{\mu}g/mL$ of each HB extract, when compared to those in the control. NO production was observed in cells pretreated with 100 or $300{\mu}g/mL$ of HBC and HBK (P < 0.01). Treatment with $300{\mu}g/mL$ of HBC significantly increased Nqo1 (P < 0.01) and Gclc (P < 0.05) mRNA levels compared to those in the control. Treatment with $300{\mu}g/mL$ of HBK (P < 0.05) and HBC (P < 0.01) also significantly increased the HO-1 mRNA level compared to that in the control. CONCLUSIONS: Thus, the Korean and Chinese HBs were found to possess favorable in vitro anti-oxidant and anti-inflammatory activities. Nrf2 and its related anti-oxidant genes were associated with both anti-oxidant and anti-inflammatory activities in HB-treated cells. Further studies are needed to confirm these in vivo effects.

Non-saponin fraction of red ginseng inhibits monocyte-to-macrophage differentiation and inflammatory responses in vitro (홍삼 비사포닌 분획의 단핵세포 분화와 염증반응에 대한 억제효과)

  • Kang, Bobin;Kim, Chae Young;Hwang, Jisu;Choi, Hyeon-Son
    • Korean Journal of Food Science and Technology
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    • v.51 no.1
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    • pp.70-80
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    • 2019
  • The aim of this study was to investigate the effects of red ginseng-derived non-saponin fraction (NSF) on inflammatory responses and monocyte-to-macrophage differentiation in RAW264.7 and THP-1. NSF effectively inhibited inflammatory responses by downregulating nitric oxide (NO) production and protein levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). NSF ($2000{\mu}g/mL$) decreased the levels of NO, iNOS, and COX-2 by 33, 83, and 64%, respectively. NSF inhibited the differentiation of monocyte-to-macrophage by decreasing cell adherence along with downregulation of the cluster of differentiation molecule $11{\beta}$ ($CD11{\beta}$) and CD36. In addition, pro-inflammatory cytokines, such as tumor necrosis factor-alpha, interleukin 6, and monocyte chemoattractant protein 1 (MCP-1), were significantly reduced with NSF treatment. The NSF-mediated inhibition of inflammatory responses was due to the regulation of nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$) and nuclear factor (erythroid-derived 2)-like 2 (Nrf2). NSF effectively suppressed the translocation of $NF-{\kappa}B$ into the nucleus, while nuclear Nrf2 and its target protein, heme oxygenase-1, levels were significantly increased.

Fermented ginseng, GBCK25, ameliorates steatosis and inflammation in nonalcoholic steatohepatitis model

  • Choi, Naeun;Kim, Jong Won;Jeong, Hyeneui;Shin, Dong Gue;Seo, Jeong Hun;Kim, Jong Hoon;Lim, Chae Woong;Han, Kang Min;Kim, Bumseok
    • Journal of Ginseng Research
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    • v.43 no.2
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    • pp.196-208
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    • 2019
  • Background: Nonalcoholic steatohepatitis (NASH) is one of the chronic inflammatory liver diseases and a leading cause of advanced liver fibrosis, cirrhosis, and hepatocellular carcinoma. The main purpose of this study was to clarify the effects of GBCK25 fermented by Saccharomyces servazzii GB-07 and pectinase, on NASH severity in mice. Methods: Six-wk-old male mice were fed either a normal diet (ND) or a Western diet (WD) for 12 wks to induce NASH. Each group was orally administered with vehicle or GBCK25 once daily at a dose of 10 mg/kg, 20 mg/kg, 100 mg/kg, 200 mg/kg, or 400 mg/kg during that time. The effects of GBCK25 on cellular damage and inflammation were determined by in vitro experiments. Results: Histopathologic analysis and hepatic/serum biochemical levels revealed that WD-fed mice showed severe steatosis and liver injury compared to ND-fed mice. Such lesions were significantly decreased in the livers of WD-fed mice with GBCK25 administration. Consistently, mRNA expression levels of NASH-related inflammatory-, fibrogenic-, and lipid metabolism-related genes were decreased in the livers of WD-fed mice administered with GBCK25 compared to WD-fed mice. Western blot analysis revealed decreased protein levels of cytochrome P450 2E1 (CYP2E1) with concomitantly reduced activation of c-Jun N-terminal kinase (JNK) in the livers of WD-fed mice administered with GBCK25. Also, decreased cellular damage and inflammation were observed in alpha mouse liver 12 (AML12) cells and RAW264.7 cells, respectively. Conclusion: Administration of GBCK25 ameliorates NASH severity through the modulation of CYP2E1 and its associated JNK-mediated cellular damage. GBCK25 could be a potentially effective prophylactic strategy to prevent metabolic diseases including NASH.

Antioxidant and anti-inflammatory activities of different extraction solvents of Cryptoporus volvatus (한입버섯의 추출 용매별 항산화 및 항염증 활성)

  • An, Gi-Hong;Han, Jae-Gu;Cho, Jae-Han
    • Journal of Mushroom
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    • v.17 no.3
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    • pp.136-143
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    • 2019
  • In this study, we analyze the antioxidant and anti-inflammatory activities of various extracts of Cryptoporus volvatus. Three extracts were prepared using hot-water, 70% ethanol, and 70% methanol. Compared to the hot-water and 70% methanol extracts, the 70% C. volvatus ethanol extracts showed significantly higher DPPH radical scavenging activities (76.1%), nitrite scavenging activities (29.5%), and total polyphenol content (9.17 mg GAE/g). The DPPH radical and nitrite scavenging activities of C. volvatus both correlated significantly with the total polyphenol content. The Pearson's correlation coefficient of DPPH radical and nitrite scavenging activities with total polyphenol content were r = 0.744 (p<0.05) and r = 0.921 (p<0.01), respectively. To test their anti-inflammatory effects, nitric oxide production and cell viability were measured by performing nitric oxide (NO) and MTT assays on lipopolysaccharide-treated RAW 264.7 cells. The NO production levels of the 70% ethanol extract-treated cells were slightly lower than those of the others. In our MTT assay, the hot-water extracts showed higher cell viabilities (81.4%) than those of 70% ethanol and 70% methanol extracts. The ${\beta}$-glucan content of C. volvatus was 37.8%. From the results of this study, it is suggested that C. volvatus, an easily accessible wild mushroom, has potential to be used in the development of medicines, cosmetics, and functional health foods.

Dammarane-type triterpene oligoglycosides from the leaves and stems of Panax notoginseng and their antiinflammatory activities

  • Li, Juan;Wang, Ru-Feng;Zhou, Yue;Hu, Hai-Jun;Yang, Ying-Bo;Yang, Li;Wang, Zheng-Tao
    • Journal of Ginseng Research
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    • v.43 no.3
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    • pp.377-384
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    • 2019
  • Background: Inflammation is widespread in the clinical pathology and closely associated to the progress of many diseases. Triterpenoid saponins as a key group of active ingredients in Panax notoginseng (Burk.) F.H. Chen were demonstrated to show antiinflammatory effects. However, the chemical structures of saponins in the leaves and stems of Panax notoginseng (PNLS) are still not fully clear. Herein, the isolation, purification and further evaluation of the antiinflammatory activity of dammarane-type triterpenoid saponins from PNLS were conducted. Methods: Silica gel and reversed-phase C8 column chromatography were used. Furthermore, preparative HPLC was used as a final purification technique to obtain minor saponins with high purities. MS, NMR experiments, and chemical methods were used in the structural identifications. The antiinflammatory activities of the isolated saponins were assessed by measuring the nitric oxide production in RAW 264.7 cells stimulated by lipopolysaccharides. Real-time reverse transcription polymerase chain reaction was used to measure the gene expressions of inflammation-related gene. Results: Eight new minor dammarane-type triterpene oligoglycosides, namely notoginsenosides LK1-LK8 (1-8) were obtained from PNLS, along with seven known ones. Among the isolated saponins, gypenoside IX significantly suppressed the nitric oxide production and inflammatory cytokines including tumor necrosis $factor-{\alpha}$, interleukin 10, interferon-inducible protein 10 and $interleukin-1{\beta}$. Conclusion: The eight saponins may enrich and expand the chemical library of saponins in Panax genus. Moreover, it is reported for the first time that gypenoside IX showed moderate antiinflammatory activity.

Comparative Study of the Antioxidant Activity Effects of Gami-Sumiwon Extract using Water and 80% Ethanol (가미수미원의 열수 추출물과 80% 에탄올 추출물의 항산화 효능 비교)

  • Lim, Hyun-Ji
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.5
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    • pp.1302-1313
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    • 2021
  • In this study, Gami-Sumiwon (GS; Cynanchi wilfordii R., Angelica gigantis R., Lycium chinense M., Betula platyphylla S., Cryptotympana atrata F. and Carthamus tinctorius L.) and/or Sinomenium acutum R. (SC) was extracted with 70% ethanol or water. And We investigated the antioxidant activity effect of GS±SC. The following experimental techniques were used to evaluate the antioxidant efficacy of GS±SC. HPLC chromatogram, heavy metal content, ABTS/DPPH radical scavenging analysis, SOD-like activity assay, FACS, and NO assay. As a result of the experiment, the sinomenine content was found to be higher in DW extracts, and decursin was found to be higher in 80% ethanol extracts. And, the amount of heavy metals in all extracts was below the standard value. ABTS, and DPPH radical scavenging activity was identified that GS±SC(EtOH) was found to have a higher scavenging activity than GS±SC(DW). But, SOD showed the opposite result. No cytotoxicity of GS was observed on Raw 264.7 cells at concentration of 1~100 ㎍/㎖. The ROS production was significantly decreased that GS±SC(DW) was found to have a higher scavenging activity than GS±SC(EtOH). However, NO production showed the opposite result. Looking at the results of SOD and ROS analysis, SC does not seem to have a function of prevention. SC is thought to have an effect on the removal of free radicals generated after oxidative stress. This result objectively confirmed the antioxidant effect of GS±SCs. We will continue to conduct in-depth research. Therefore, it is believed that the possibility of using GS±SCs as a functional material can be established. The more diverse the objectives, the higher the value of GS utilization is thought to be.