• Title/Summary/Keyword: In vivo와 In vitro

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Primary Culture and Characteristics of Blood-Brain Barrier Endothelial Cells from Rat Brain Microvessel (쥐의 뇌 미세혈관 내피세포를 이용한 뇌혈관장벽 내피세포의 배양과 특성)

  • Lee, Hee-Sang;Kim, Seok-Jung;Kim, Dae-Jin;Chung, Yoon-Hee;Kim, Sung-Su;Lee, Won-Bok;Kim, Kyung-Yong
    • Applied Microscopy
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    • v.35 no.4
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    • pp.55-63
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    • 2005
  • The characteristics of primary cultured rat brain microvessel endothelial cells (RBMECs) were studied using microscopy, immunohistochemistry and measuring of transendothelial electrical resistance (TER). The RBMECs formed a monolayer by $5{\sim}6$ days after plating and showed characteristics of whirling appearance. The TER increased until day 5 and decreased then. There was few immunoreaction with anti-GFAP, anti-GalC, anti-neurofilament 160/200 kD antibodies. So the contamination of astrocyte, oligodendrocyte, and neuron. could be ruled out.. Immunoreaction to vWF antigen was widespread througout the cytoplasm as Weibel-Palade granule. Immunoreaction to tight junction proteins, i.e. occludin, ZO-1, and ZO-2 was seen at cell contact. In summary, RBMECs isolated and cultured showed morphological, immunohistochemical and electrical characteristics of blood-brain barrier (BBB). The in vitro BBB model can be used in studying characteristics of in vivo BBB.

Various physiological effects from fruiting body extracts of Phellinus baumii (장수진흙버섯 자실체 추출물의 다양한 생리활성 효과)

  • Yoon, Ki Nam;Lee, Tae Soo
    • Journal of Mushroom
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    • v.18 no.3
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    • pp.260-267
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    • 2020
  • Phellinus baumii, a white-rot fungus, has been used for centuries as folk medicine in China, Japan, and Korea. This study aimed to evaluate the in vitro anti-diabetic, and anti-cholinesterase, and in vivo anti-inflammatory effects of the fruiting bodies of P. baumii. The methanol (ME) and hot water (HE) extracts (2.0 mg/mL) of P. baumii fruiting bodies suppressed α-amylase activity, exactly 61.33%, and 65.00%, respectively; of note, acarbose, the positive control, inhibited 93.33% of the α-amylase activity. Moreover, the ME and HE (2.0 mg/mL) inhibited 89.67% and 91.00%, respectively, of the activity of α-glucosidase activity, whereas the same concentration of acarbose suppressed 84.67% of the α-glucosidase activity. The ME and HE (1.0 mg/mL) also inhibited 96.05% and 94.58%, respectively, of the acetylcholinesterase (AChE) activity; galanthamine, the positive control, led to an inhibition of 81.12%. The butyrylcholinesterase (BChE) activity was also inhibited by ME and HE (1.0 mg/mL; 91.05% and 82.27%, respectively); of note, the same concentration of galanthamine suppressed 81.12% of the BChE activity. The production of NO in LPS-induced RAW 264.7 macrophages was significantly suppressed by both ME and HE treatments. Importantly, the carrageenan-activated rat hind-paw edema was significantly reduced 2-6 h after ME administration (50 mg/mL). Taken together, the results suggest that the fruiting bodies of P. baumii have α-amylase, α-glucosidase, α-cholinesterase, and anti-inflammatory activities, and, therefore, may be good natural sources for the promotion of human health.

Inhibitory Effects of Several Korean Traditional Herbs on Anaphylactic Reaction and Mast Cell Activation (아나필락시스 반응과 비만세포 활성화에 대한 한국전통약제의 억제효과)

  • Chai, Ok-Hee
    • Applied Microscopy
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    • v.40 no.4
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    • pp.201-209
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    • 2010
  • Korean traditional herbs, especially Anemarrhena asphodeloides (A. asphodeloides), Salvia miltiorrhiza (S. miltiorrhiza), and Terminalia chebula (T. chebula) have been known to have the immunomodulatory, anti-tumor, and anti-inflammatory effects. However, direct cellular mechanism underlying the mast cell-mediated anaphylaxis-like reaction has poorly been understood. In the present study, the effects of the methanol extracts of A. asphodeloides (MEAA), S. miltiorrhiza (MESM), and T. chebula (METC) on anaphylactic reaction were investigated. Using in vitro and in vivo experiments, the influences of MEAA, MESM, and METC on the compound 48/80-induced anaphylaxis-like reaction and mast cell activation, and IgEmediated PCA were examined. Results are below; 1) MEAA, MESM, and METC significantly inhibited systemic anaphylaxis-like reaction, ear swelling response, and IgE-mediated PCA. 2) the compound 48/80-induced mast cell degranulation, histamine release of rat peritoneal mast cells (RPMC) were significantly inhibited by the pretreatment with MEAA, MESM, and METC, and 3) the compound 48/80-induced calcium influx in RPMC was significantly inhibited by the pretreatment with MEAA, MESM, and METC. These results suggest that MEAA, MESM, and METC may be an activity to inhibit the compound 48/80-induced or anti-DNP IgE-mediated anaphylactic reactions by blocking histamine release and calcium uptake into RPMC. MEAA, MESM, and METC potentially may serve as an effective therapeutic tool for allergic diseases.

Establishment and Characterization of Canine Mammary Gland Cancer Cell Lines (개 자연발생 유선종양 2종의 세포주 확립 및 특성분석)

  • Lee, Sun-Tae;Kweon, Oh-Kyeong;Kim, Wan-Hee
    • Journal of Veterinary Clinics
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    • v.27 no.3
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    • pp.232-239
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    • 2010
  • Two cell lines derived from spontaneous canine mammary gland tumors were established and characterized. Mammary gland tumors from 9 years old pug and 9 years old toy-poodle dogs were collected by aseptic surgical resection and primary culture was performed. The histopathologic examination of tumors revealed adenocarcinoma and complex carcinoma and two dogs died from metastasis of the tumors. The tumor cells were subcultured over 60 times for more than 1 year and morphological consistency maintained. Light microscopic examination, growth curve, doubling time calculation, xenotransplantation to female nude mice, immunohistochemistry for wide spectrum keratin, vimentin, $\alpha$-smooth muscle actin and cytokeratin 8 was performed for characterization. The cell lines exhibited polygonal, elongated cell shape and cytoplasmic bridge and doubling time of 47.1 hrs and 18.6 hrs, respectively. Subcutaneous xenotransplantation to nude mice of the cells produced localized palpable mass within 4 weeks in 4 of 5 and 5 of 5 nude mice, respectively. In immunohistochemical examination one cell line showed strong positive against wide spectrum keratin and cytokeratin 8 and the other cell line showed strong positive against smooth muscle actin and cytokeratin 8. Additional characterization would be possible by investigator's needs and the cell lines may be useful for in vivo and in vitro studies of canine mammary tumor and adjuvant therapies.

Interleukin-5 Inhibition Assay of the Oriental Materia Medica Treatment by High Through Screening on the Splenocyte of Mouse (High through screening(HTS)를 이용한 백서(白鼠)의 비장세포(脾臟細胞)에서의 IL-5 분비 조절 한약물(韓藥物) Screening)

  • Park, Dong-Hee;Rhee, Hyung-Koo;Jung, Sung-Ki;Jung, Hee-Jae
    • The Journal of Internal Korean Medicine
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    • v.27 no.2
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    • pp.394-406
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    • 2006
  • Backgrounds and Objectives: Asthma is considered to be an inflammatory disease characterized by airway hyperresponsiveness and Pulmonary eosinophilia. And it is known the structure and function of IL-5, its receptor and the mechnism IL-5 triggered eosinophil accumulation and inflammaion of the airways. At this point of view, we assume which oriental materia medics can the splenocyte inhibit from secreting the IL-S in vitro. Material and Methosds: We used the splenocyte of mouse 8 weeks after its birth, and then cnltivated those into the 2 experimental groups and control group for 48 hours. The culture medium of experimental groups were made of $1{\mu}g/ml,\;10{\mu}g/ml$, oriental materia medics, representative. And the culture media of control group was given no oriental materia medica. Then, we assayed the quantity of cytokine-expression by the Sandwich ELISA. The quantifies of cytokine-expression of the experimental groups were compared with that of the control group which was standardized These method were used for the all of oriental materia medica treated. Results: In this study, we demonstrated that 12 oriental materia medica that inhibit the splenocyte from secreting the IL-5 in both $1{\mu}g/ml,\;and\;10{\mu}g/ml$ culture media. Those were Equiseti Herbs, Sophorae Subprostratae Radix, Moutan Radicis Cortex. Trichosanthis Radix, Buddleiae Flos. Cyperi Rhizoma. Benincasae Semen, Armeniacae Semen. Zedoariae Rhizoma, Astragali Semen, Dolichoris Semen. Lilii Bulbus, Asparagi Radix, Atractylodes Rhizoma White, Polygonati Officinallis Rhizoma. Conculusions: These findinga indicate that some oriental materia medica, specially Antipyretics, Herbs for Resolving Phlegm, Relieving Cough and Calming Wheezing and Herbs for Tonifring and Invigorating effects inhibit the splenocyte from secreting the IL-5. And further study experimented in vivo is needed for treating IL-5-driven inflammatory disease including asthma.

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Anti-oxidative Activity and the Protective Effect of Donkey's Bone and Skin Extracts on SK-N-SH Cells (당나귀 사골과 껍질의 항산화기능 및 SK-N-SH세포 보호효과)

  • Kim, Dongwook;Chae, Hyun-Seok;Kim, Nam-Young;Jang, Aera
    • Journal of Life Science
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    • v.23 no.8
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    • pp.1019-1024
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    • 2013
  • The aims of this study were to determine antioxidation effect and neuroblastoma cell protection effect of donkey's bone and skin extracts (DBSE). DBSE was extracted by a pressure-cooker for 48 h and lyophilized. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity was significantly increased with increased doses of DBSE and 40 mg/ml of DBSE showed 95.43% of the DPPH scavenging effect, which was equivalent to 1 mg/ml of vitamin C. The 2,2'-azino-bis (3-ethylbenzothiazoline-6- sulphonic acid) (ABTS) radical scavenging activity was also increased in a dose-dependent manner, and 20 mg/ml of DBSE showed 88.73% of the ABTS scavenging effect. The oxygen radical absorbance capacity (${\mu}M$ Trolox equivalent) of DBSE was significantly increased at a concentration of 10 mg/ml, which showed $132.53{\mu}M$ TE. The viability of oxidatively stressed brain cells induced by $500{\mu}M\;H_2O_2$ was protected by DBSE at concentrations greater than $50{\mu}M$. Cell viability after DBSE treatment at 50 and $100{\mu}g/ml$ was 53.78 and $54.34{\mu}M$ TE, respectively. There was no significant difference between both doses; however, 200 and $500{\mu}g/ml$ of DBSE showed 59.74 and 66.08% of cell viability, respectively indicating that DBSE protected SK-N-SH from oxidation stress. These results suggest that DBSE may have potential to be used as natural antioxidants in food industry, while in vivo evidence is necessary to support DBSE's in vitro-based antioxidative efficiency.

Immunostimulating and Anticancer Activities of Hot Water Extract from Capsosiphon fulvescens (매생이 열수추출물의 면역 및 항암 활성)

  • Park, Hee-Yeon;Lim, Chi-Won;Kim, Yeon-Kye;Yeon, Ho-Dong;Lee, Ka-Jung
    • Applied Biological Chemistry
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    • v.49 no.4
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    • pp.343-348
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    • 2006
  • To investigate the immunostimulating and anticancer activities from hot water extract of Capsosiphon fulvescens, tumor cell toxicity, sarcoma-180 growth inhibition activity, complement system-activity, intestinal immune system and oral toxicity were performed. The extract of Capsosiphon fulvescens was prepared by hot water and precipitated by using ethanol. Partially purified extract (CFE) was obtained after dialysis and ultrafiltration. The polysaccharide compositions consisted of xylose(19.1%), fucose(15.3%), mannose(4.2%) and galactose(7.9%). The tumor cell toxicity of CFE slightly showed at high concentrations of 10-30 ${\mu}g/ml$, but inhibition ratio against mouse solid tumor was more increased for CFE of 40.1-59.4% than the control. Blood leukocyte counts increased to a maximum of 83% including liver, spleen and thymic of mouse. Immunoglobulin A binding amounts showed a high level of CFE of $2,454{\pm}113.8-2,670{\pm}133.1{\mu}g/mg$ in comparison with the control of $2,092{\pm}123.0{\mu}g/mg$. Acute toxicity of CFE was not detected at the concentration of 2,000 mg/kg in normal mouse.

Antioxidative and Anti-diabetic Activity of C3GHi, Novel Black Rice Breed (Cyanidine-3-glucoside (C3G) 색소의 함유량이 증대된 쌀 신품종(C3GHi)의 항산화 및 항당뇨 활성)

  • Kim, Hwa-Young;Kim, Joong-Hark;Lee, Sung-Ae;Ryu, Su-Noh;Han, Sang-Jun;Hong, Seong-Gil
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.55 no.1
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    • pp.38-46
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    • 2010
  • C3GHi, a novel black rice variety developed by conventional breeding has more high contents of cyanidine-3-glucoside (C3G) and a more strong antioxidant than normal black rice. In this study, we investigate the antioxidative potential and anti-diabetic activity of C3GHi and extract of it for the purpose of development of functional materials or replacement meal for diabetic patients. Using in vitro antioxidative activity assay system such as DPPH radical quenching assay, superoxide anion radical scavenging assay, inhibition of lipid peroxidation and DNA breakage assay, C3GHi extract was observed to have more high antioxidative activity than normal black rice breed. The glycemic index of a freeze dried powder of C3GHi brown rice showed $43.7\pm18.8$, which belonged to low GI food (GI of 55 or less). Using in vivo diabetic model such as db/db mice model and streptozotocininduced diabetic model, C3GHi rice extract decrease blood glucose level and inhibit oxidative stress in blood. From the results, we think that the C3GHi varieties have a potential for the functional materials or ingredient of meal replacement for diabetic patients.

Microgrooves on titanium surface affect peri-implant cell adhesion and soft tissue sealing; an in vitro and in vivo study

  • Lee, Hyo-Jung;Lee, Jaden;Lee, Jung-Tae;Hong, Ji-Soo;Lim, Bum-Soon;Park, Hee-Jung;Kim, Young-Kwang;Kim, Tae-Il
    • Journal of Periodontal and Implant Science
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    • v.45 no.3
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    • pp.120-126
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    • 2015
  • Purpose: With the significance of stable adhesion of alveolar bone and peri-implant soft tissue on the surface of titanium for successful dental implantation procedure, the purpose of this study was to apply microgrooves on the titanium surface and investigate their effects on peri-implant cells and tissues. Methods: Three types of commercially pure titanium discs were prepared; machined-surface discs (A), sandblasted, large-grit, acid-etched (SLA)-treated discs (B), SLA and microgroove-formed discs (C). After surface topography of the discs was examined by confocal laser scanning electron microscopy, water contact angle and surface energy were measured. Human gingival fibroblasts (hGFs) and murine osteoblastic cells (MC3T3-E1) were seeded onto the titanium discs for immunofluorescence assay of adhesion proteins. Commercially pure titanium implants with microgrooves on the coronal microthreads design were inserted into the edentulous mandible of beagle dogs. After 2 weeks and 6 weeks of implant insertion, the animal subjects were euthanized to confirm peri-implant tissue healing pattern in histologic specimens. Results: Group C presented the lowest water contact angle ($62.89{\pm}5.66{\theta}$), highest surface energy ($45{\pm}1.2mN/m$), and highest surface roughness ($Ra=22.351{\pm}2.766{\mu}m$). The expression of adhesion molecules of hGFs and MC3T30E1 cells was prominent in group C. Titanium implants with microgrooves on the coronal portion showed firm adhesion to peri-implant soft tissue. Conclusions: Microgrooves on the titanium surface promoted the adhesion of gingival fibroblasts and osteoblastic cells, as well as favorable peri-implant soft tissue sealing.

Anti-wrinkle Effect of Cosmetics Containing Duchesnea indica Extract (사매추출물을 함유하는 화장품의 주름 개선 효과)

  • Yang, Woong-Suk;Kim, Young-Min;Kim, Ee-Hwa;Seu, Young-Bae;Yang, Yoon-Jung;Kim, Hyun-Woo;Kang, Se Chan
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.36 no.4
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    • pp.281-288
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    • 2010
  • In this study, we investigated anti-oxidative effects of Duchesnea indica extracts by using Oxygen Radical Absorbance Capacity (ORAC). The extracts were prepared with 0 %, 30 %, 50 %, 70 % and 100 % aqueous ethanol respectively. The 30 % EtOH D. indica extract showed higher ORAC activity than the other extracts. Therefore, we performed in vitro studies on cytotoxicity of NIH-3T3 cells and MMP-8 collagenase inhibition using by the 30 % EtOH extract. The 30 % EtOH extract showed no cytotoxicity and significant inhibition on MMP-8 collagenase. And we performed clinical studies for the anti-wrinkle effect of the Di-Wrinkle Free Cream. The cream formula was prepared with 2 % arbutin and 1 % D. indica extract. Twenty one healthy women volunteers, ages of 35 and 50, applied the cream on their faces twice a day for 8 weeks. The skin was evaluated with PRIMOS (phaseshift rapid in vivo measuring of human skin) system and analyzed by the student's paired t-test. The wrinkles on the eye region were reduced by 13 % based on the PRIMOS system after 8 weeks. In the safety study of the Di-Wrinkle Free Cream, no symptoms were observed such as erythema, edema, scaling, itching, stinging, burning, tightness and prickling by visual observation and medical examination of volunteers for 8 weeks. Moreover, there was no noticeable skin disorder during experience period. These results suggested that D. indica extracts could be applied as cosmeceuticals effective for anti-wrinkle.