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Relationship between the Regulator of Calcineurin 1-4 Isoform and In Vitro Osteoclast Differentiation (Regulator of calcineurin 1-4과 파골세포 분화의 관련성)

  • Park, Kyeong-Lok
    • Journal of Life Science
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    • v.25 no.2
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    • pp.223-230
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    • 2015
  • Regulator of calcineurin 1 (RCAN1) is an endogenous calcineurin inhibitor that plays an important role in the pathogenesis of diseases related to the calcineurin-NFATc1 signaling pathway. The RCAN1-4 isoform is subject to NFATc1-dependent regulation. During receptor activator of nuclear factor kappa-B ligand (RANKL)-stimulated osteoclastogenesis, the calcineurin-NFATc1 pathway is critical. Because there is little information available on the role of RCAN1 in osteoclast differentiation, this study investigated whether changes in RCAN1 expression are related to the calcineurin-NFATc1 pathway and osteoclast differentiation. Mouse bone marrow monocytes (BMMs) were treated with 50 ng/ml of RANKL and M-CSF. Expression levels of NFATc1, calcineurin, and RCAN1 isoforms were determined using RT-PCR and Western blotting. Osteoclast differentiation was examined using tartrate-resistent acid phosphatase (TRAP) staining. To evaluate the effect of RCAN1 overexpression on osteoclastogenesis, cells were transfected with a mouse RCAN1-4 cDNA plasmid. After RANKL stimulation of BMMs, expression of NFATc1 and RCAN1 was increased at the mRNA and protein level, while calcineurin expression was unchanged. When the RCAN1-4 gene construct was transfected, the expression of RCAN1 protein was not increased despite several-fold increases in RCAN1-4 mRNA expression. Regardless of RANKL stimulation, over-expression of RCAN1-4 tended to reduce NFATc1 expression and knock-down of RCAN1 increase it. While BMMs transfected with the RCAN1-4 vector were differentiated into distinct osteoclasts, their phenotypes did not vary from those of mock controls. These results suggest that RCAN1 has a limited effect on the calcineurin-NFATc1 pathway during RANKL-stimulated osteoclast differentiation.

Antioxidant and Anti-Inflammatory Activities of Extracts of Traditional Medicinal Plants Mixtures 1 and 2 (약용식물 복합 추출물 TMP1과 TMP2의 항산화능과 염증 억제 효과)

  • Cho, Eun-Kyung;Choi, Young-Ju
    • Journal of Life Science
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    • v.21 no.4
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    • pp.596-603
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    • 2011
  • This study was conducted to investigate traditional medicinal plants (TMP) 1 and 2, two different multi-herbal mixtures consisting of 24 herbs. Regarding the contents of flavonoid compounds, the ethanol extract (EE) of TMP2 yielded the highest content of flavonoid compounds (40.981 mg/g), followed by EE of TMP1 (28.23 mg/g), hot water extract of TMP2 (WE, 10.046 mg/g), and WE of TMP1 (6.59 mg/g). Antioxidant activities of EE and WE of TMP1 and TMP2 were measured based on DPPH radical scavenging activity. At 1 mg/ml, the highest DPPH radical scavenging activity was shown in EE of TMP2 (96%), followed by EE of TMP1 (94%). Nitric oxide (NO) production by RAW264.7 macrophage cells stimulated with lipopolysaccharide (LPS) was reduced to 82, 77, 86, and 47% by addition of $500\;{\mu}g/ml$ of EE and WE of TMP1 and TMP2, respectively. These results were not due to the cytotoxicity of the extracts. NO synthesis was increased by 2.3, 3.6, 3.0, and 1.8 fold by addition of $500\;{\mu}g/ml$ of EE and WE of TMP1 and TMP2, respectively. These results show that medicinal plants play a significant role in antioxidative activity and activation of the immune system in the pathogenesis of inflammatory diseases, and different mixtures of the medicinal plants showed different effects.

Antioxidant, Antidiabetic, and Anti-Inflammatory Effects of Extracts and Fractions from Parthenocissus tricuspidata Stems (담쟁이덩굴 추출물과 분획물의 항산화, 항당뇨 및 항염증 효과)

  • Cho, Eun Kyung;Choi, Young Ju
    • Journal of Life Science
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    • v.23 no.3
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    • pp.399-405
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    • 2013
  • This study was carried out to analyze the effects on antioxidative, antidiabetic, and anti-inflammatory activities of Parthenocissus tricuspidata (PT) stem extracts. The total phenolic contents of hot water and ethanol extracts from PT stems were 61.5 mg TAE/g and 122.1 mg TAE/g, respectively. The antioxidative activities of hot water and ethanol extracts from PT stem were measured by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and superoxide dismutase (SOD) assay. The DPPH radical scavenging activities of ethanol extract and butanol fraction were approximately 95% and 92% at 1 mg/ml, respectively, and the SOD activities of ethanol extract and butanol fraction were about 91% and 97% at 1 mg/ml, respectively. The DPPH radical scavenging and SOD activities of ethanol extract and butanol fraction from PT stem increased remarkably increased in a dose-dependent manner and were higher than in the hot water extracts. Compared to the acarbose, a known anti-diabetic drug, which was used as a positive control, the ${\alpha}$-glucosidase inhibitory capacity of PT stem showed a strong inhibitory rate in ethanol extract and in butanol and hexane fractions. We investigated the effect of hot water extract from PT stem on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 cells. Hot water extract from PT stem inhibited LPS-induced NO production up to 40% at a treatment of 1 mg/ml. These results suggest that PT stem extracts have an effect on antioxidative, antidiabetic, and anti-inflammatory activities and thus have great potential as antidiabetic materials and a source for natural health products.

Anti-inflammatory Effect of Onion (Allium cepa) Peel Hot Water Extract in vitro and in vivo (양파껍질 열수추출물의 in vitro 및 in vivo 항염증 효과)

  • Kang, Bo-Kyeong;Kim, Koth-Bong-Woo-Ri;Ahn, Na-Kyung;Choi, Yeon-Uk;Kim, Min-ji;Bark, Si-Woo;Pak, Won-Min;Kim, Bo-Ram;Park, Ji-Hye;Bae, Nan-Young;Ahn, Dong-Hyun
    • KSBB Journal
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    • v.30 no.4
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    • pp.148-154
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    • 2015
  • Onion (Allium cepa) is one of the flavonoids-rich materials in human diet and onion peel, which is the onion by-products, contains over 20 times more quercetin than the flesh. In this study, to examine the anti-inflammatory effects of onion peel hot water extract (OPHWE), the cell viability, nitric oxide (NO), pro-inflammatory cytokines, such as interluekin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and IL-$1{\beta}$, were measured using the murine macrophage cell line RAW 264.7 cells. The Balb/c mice were used for an in vivo acute toxicity test and ICR mice were used for measurement of inhibition effects of croton oil-induced mouse ear edema. As a result, NO levels decreased in a dose-dependent manner. The production of IL-6, TNF-${\alpha}$, and IL-$1{\beta}$ was suppressed by 38%, 41%, and 34% respectively, compared with that of the LPS only group, without any cytotoxicity. The edema formation in the ICR mouse ear was also reduced compared to that in control. Moreover, there were no mortalities occurred in mice administered 5,000 mg/kg body weight of OPHWE. These results suggest that OPHWE has considerable anti-inflammatory activities and can be regarded as a potent candidate material to treat inflammatory diseases.

Analysis of Immunomodulating Activities in Methanol Extracts from Several Kinds of Grapes (포도 품종별 메탄올 추출물로부터 면역활성 분석)

  • Heo, Jin-Chul;Woo, Sang-Uk;Kweon, Mi-Ae;Kim, Bo-Bae;Lee, Sook-Hee;Lee, Jin-Man;Choi, Jong-Uck;Chung, Shin-Kyo;Lee, Sang-Han
    • Food Science and Preservation
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    • v.14 no.4
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    • pp.419-424
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    • 2007
  • Fruits and vegetable extracts are well-known as healthy foods. Such foods have been used as herbal medicines or traditional therapies for centuries. To assess biological activities in grapes, we examined the immunomodulating activities of water extracts from four kinds of grapes (cultivars Kyoho, Delaware, Campbell, and Niagara). We explored possible antioxidant and anticancer activities using antioxidant assays such as the 1,1-diphenyl-2-picrylhydrazyl (DPPH) reduction assay, the ferric iron reducing ability of plasma (FRAP) assay, a cell proliferation assay, an NO inhibition assay, a wound healing assay, and an IL-4/IL-13 elicitation assay. Methanol extracts of grapes were tested. The results showed that each grape extract had potent antioxidant activities. The grape extracts increased cell proliferation and NO production activities in tumor cell lines. IL-4 and IL-13 cytokine levels were decreased in mouse primary spleen cells by treatment with any extract. These results suggest that grape extracts can be used as biomaterials with immunomodulating activities.

Comparison of Antioxidant, Anticancer and Immunomodulating Activities of Extracts from DongChongXiaCao (동충하초의 균사 및 배양액의 항산화, 항암, 면역활성의 비교)

  • Heo, Jin-Chul;Nam, Sung-Hee;Kang, Seok-Woo;Hong, In-Pyo;Lee, Kwang-Kil;Park, Ja-Young;Kim, Kyung-Hae;Han, Song-Yi;Lee, Sang-Han
    • Food Science and Preservation
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    • v.14 no.6
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    • pp.681-687
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    • 2007
  • Biological properties of DongChongXiaCao extracts and culture supernatants were evaluated using DPPH and FRAP (antioxidants), Raw 264.7 (NO production), B16-F1 cells (cell migration activity) and HUVECs (angiogenesis activity). We found that antioxidant activity was higher in mycelium culture supernatants than in mycelial extracts. Mycelial extracts and culture supernatants inhibited or increased cyclooxygenase-2 transcription activity and NO production. Various extracts and culture supernatants inhibited B16 cell migration and motility, and inhibited HUVEC tube formation. These findings indicate that DongChongXiaCao extracts and products of mycelium could be a useful biological resource for anti-oxidant and anti-cancer purposes.

Anti-atopic Activity of Sargassum micracanthum Ethanol Extracts (잔가시 모자반 에탄올 추출물의 항아토피 효과)

  • Jeong, Da-Hyun;Kim, Koth-Bong-Woo-Ri;Kim, Min-Ji;Kang, Bo-Kyeong;Bark, Si-Woo;Pak, Won-Min;Kim, Bo-Ram;Park, Hong-Min;Im, Moo-Hyeog;Ahn, Dong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.42 no.1
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    • pp.82-88
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    • 2014
  • Atopic dermatitis (AD) is a common chronic inflammatory disease preceding the development of allergic disorders. The aim of this study was to evaluate the effect of Sargassum micracanthum ethanol extract (SMEE) on AD. AD was induced by spreading 2,4-dinitrochlorobenzen (DNCB) on the back sides of BALB/c mice. The efficacy of SMEE was tested by observing the skin clinical severity score, proliferations of Raw 264.7 cells and the secretion of cytokines and IgE. The secretion of IL-4, and IgE was significantly decreased by SMEE in a dose dependent manner, while IFN-${\gamma}$ was increased. In addition, SMEE alleviated the AD symptoms better when compared to the positive controls. In conclusion, these results suggest that SMEE has an inhibitory effect on AD, and may serve as a useful biomaterial for the development of cosmeceuticals.

Hypolipidemic and Anti-oxidant Effects of Chunghyl Plus in Type II Diabetic Mice Model (제2형 당뇨 마우스 모델에서 청혈플러스의 항고지혈 및 항산화효과)

  • Choi, Koh Eun;Seol, In Chan;Kim, Yoon Sik;Cho, Hyun Kyoung;Yoo, Ho Ryong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.30 no.3
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    • pp.164-176
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    • 2016
  • This study was perfomed to investigate the effects of Chunghyul-plus(CHP) on oxidative damage and hyperlipidemia in db/db mouse. After treatment with CHP, safety in cytotoxicity, heavy metal toxicity, production of reactive oxygen species(ROS), nitric oxide (N0) and proinflammatory cytokine IL-Ib, TNF-a, IL-6 in RAW 264.7 cells. Serum total cholesterol, LDL cholesterol, HDL cholesterol, triglyceride, insulin, GLP-1, glucose, food intake, body weight, organ weight, AST, ALT, ALP, BUN, creatine and histologic change of liver and aorta were measured in db/db mouse after oral administration of CHP. CHP showed safety in cytotoxicity and toxicity of liver and kidney for logn time administration. CHP increased the DPPH and ABTS radical scavenging activity. CHP showed significant inhibitory effect on reactive oxygen species (ROS), and showed inhibitory effect on nitiric oxide(NO) compared to control group. CHP decreased cytokine IL-6 production significantly, and decreased IL-1β and TNF-α compared to control group. CHP decreased body and organ weitht, intake food, and glucose levels compared to control group. CHP decreased total cholesterol and triglyceride significantly, and decreased LDL-cholesterol levels and increased HDL-cholesterol levels compared to control group. CHP decreased atherogenic index and cardiac risk factor significantly. CHP increased serum insulin and GLP-1 compared to control group. In histologic examination, lipophagy in the liver and aorta decreased in CHP treated mice and the cell was regular and boundary of vessel wall was clear compared to control group. These results suggest that CHP is effective in antioxidation activity and treatment and prevention of hyperlipidemia, atherosclerosis, diabetes, ischemic heart disease, stroke and other cardiocerebrovascular disease.

Composition Comprising the Extract of Anethi Fructus for the Treatment and Protection of Immune Activity (시라자 추출물을 함유하는 면역질환의 치료 및 예방을 위한 면역증강용 조성물)

  • Park, Gil-Soon;Chang, In-Ae;Kim, Youn-Chul;Lee, Moo-Hyung;Shin, Hye-Young;Choi, Du-Young;Yun, Yong-Gab;Park, Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.3
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    • pp.700-704
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    • 2007
  • In the recent, increased concern has been focused on the pharmacology and clinical utility of herbal extracts and derivatives as a drug or adjunct to chemotherapy and immunotherapy. Here we investigated the role of the extract of Anethi Fructus in the expression of inflammatory mediators, surface molecule, and related receptors in vitro. In murine macrophage RAW 264.7 cells and peritoneal macrophages of C57BL/6N mice, water extract of Anethi Fructus increased the production of secretary tumor necrosis factor (TNF)-a and Nitric oxide (NO), and the expression level of CD14, LPS co-receptor and CD86, co-stimulatory molecule compared to negative natural extract ex vivo. The water extract of Anethi Fructus increased the production of interferon (IFN)-g from splenocytes. Also, water extract of Anethi Fructus increased ConA-induced cell proliferation. These results suggest that water extract of Anethi Fructus may enhance the immune response through immune modulation of macrophage and lymphocytes.

A Comparison between Water and Ethanol Extracts of Rumex acetosa for Protective Effects on Gastric Ulcers in Mice

  • Bae, Ji-Yeong;Lee, Yoon-Seok;Han, Sun-Young;Jeong, Eun-Ju;Lee, Mi-Kyeong;Kong, Jae-Yang;Lee, Dong-Hoon;Cho, Kyeong-Jae;Lee, Haeng-Soon;Ahn, Mi-Jeong
    • Biomolecules & Therapeutics
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    • v.20 no.4
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    • pp.425-430
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    • 2012
  • Rumex acetosa is a perennial herb that is widely distributed across eastern Asia. Although the hot water extract of R. acetosa has been used to treat gastritis or gastric ulcers as a folk medicine, no scientific report exists for the use of this plant to treat gastric ulcers. Hence, the present study was undertaken to assess the anti-ulcer activity of water and 70% ethanol extracts obtained from R. acetosa, using an HCl/ethanol-induced gastric ulcer model in mice. Anti-inflammatory and free radical-scavenging activities of these two extracts were also evaluated and compared. As a result, the administration of R. acetosa extracts significantly reduced the occurrence of gastric ulcers. However, significant differences in protective activity against gastric ulcers were observed between the two samples. In the case of the group pretreated with an ethanol extract dosage of 100 mg/kg, the protective effect (90.9%) was higher than that of water extract (41.2%). Under histological evaluation, pretreatment with R. acetosa extracts reversed negative effects, such as inflammation, edema, moderate hemorrhaging and loss of epithelial cells, presented by HCl/ethanol-treated stomachs. Meanwhile, R. acetosa extracts showed potent DPPH radical-scavenging activity and decreased NO production in a murine macrophage cell line, RAW 264.7, in a dose-dependent manner without affecting cellular viability. The greater anti-ulcer and NO production inhibitory activities exhibited by ethanol extracts compared to water extracts could be ascribed to the higher emodin levels, a major anthraquinone component of this plant.