• Title/Summary/Keyword: MTT method.

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Effects of Scutellariae Radix-acupuncture solution On Anti-inflammatory in RAW 264.7 macrophages (황금약침액(黃芩藥針液)이 LPS로 유도(誘導)된 RAW 264.7 대식세포(大食細胞)에서의 항염증효과(抗炎症效果))

  • Choi, Young-Kwang;Park, Sung-Ho;Seo, Il-Bok;Kim, Ho-Hyun;Kim, Jeong-Seon;Kim, Ee-Hwa
    • Journal of Pharmacopuncture
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    • v.6 no.3
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    • pp.5-14
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    • 2003
  • Objective : Recently, Herbal-acupuncture therapeutics has been used for the treatment of inflammatory diseases such as rheumatoid arthritis. Especially, we have been interested in chemical mediators concerned with inflammation such as prostaglandin, cytokine, nitric oxide. The purpose of this study is investigated that the effect of Scutellariae Radix-acupuncture solution in lipopolysaccharide-stimulated RAW 264.7 macrophages, performed several expeimental items : those are prostaglandin E. nitric oxide and cyclooxygenase-2. Method : The cytotoxicity of Scutellariae Radix-acupuncture solution in RAW 264.7 macrophages were measured by MTT-based cytotoxicity assay. In order to observe cyclooxygenase-2 mRNA expression in lipopolysaccharide-stimulated RAW 264.7 macrophages, RT-PCR was used. Prostaglandin $E_2$ formation and nitric oxide production was measured by competitive enzyme immunoassay and Griess assay. Results : 1. The MTT assay demonstrated that cytotoxic effect of Scutellariae Radix-acupuncture solution in RAW 264.7 macrophages were not appeared before concentration of 5mg/mL 2. Scutellariae Radix-acupuncture solution inhibited cyclooxygenase-2 mRNA expression in lipopolysaccharide-treated RAW 264.7 macrophages. 3. Scutellariae Radix-acupuncture solution inhibited nitric oxide production in lipopolysaccharide-treated RAW 264.7 macrophages. 4. Scutellariae Radix-acupuncture solution inhibited prostaglandin $E_2$ formation in lipopolysaccharide-treated RAW 264.7 macrophages. Conclusion : On the basis of these results, It was shown that Scutellariae Radix-acupuncture solution is significantly able to inhibit the production of $PGE_2$ and NO, as well as COX-2 mRNA expression. Our results may provide new mechanism by which Scutellariae Radix-acupuncture solution accounts for its beneficial effect on accelerating wound healing and anti-inflammation.

Quality Properties and Anti-allergic Effect of Makgeolli Added with Garlic (마늘 첨가 막걸리의 품질특성 및 항알레르기 효과)

  • Ko, Yu-Jin;Kang, Sang-Dong;Kang, Sang-Tae;Ryu, Chung-Ho
    • Journal of Life Science
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    • v.21 no.11
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    • pp.1592-1598
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    • 2011
  • Allium sativum, commonly known as garlic, is well known for its antibacterial, antioxidant, and hypotensive activities, as well as its antiaging bioactive properties. This study investigated the physicochemical properties and anti-allergic effects of garlic in order to develop a method to produce Makgeolli using garlic. Levels of total sugar, reducing sugar contents, total acid and amino acid contents of garlic Makgeolli were higher than those of the control (normal Makgeolli without garlic), and its alcohol content was 7.0%. Polyphenol contents, DPPH (1, 1-diphenyl-2-picryl-hydrazyl) free radical scavenging activity and total thiosulfate contents of garlic Makgeolli were a bit higher than those of the control. The main organic acids of garlic Makgeolli were lactic, citric, malic, oxalic and succinic acids. Especially, lactic acid content was 16 fold higher than the control. According to MTT assay, garlic Makgeolli had no significant effect on the viability of human mast cells, and secretion of histamine was more effectively inhibited by garlic Makgeolli than by the control. Garlic Makgeolli was superior to the control in all results of sensory characteristics except in color. Therefore, the anti-oxidant activity and anti-allergic effect of garlic Makgeolli could improve the quality of Korean Makgeolli.

Anti-oxidation and Anti-atopic Dermatitis Effect of Herbal Wood Vinegar (한방목초액의 항산화 및 항 아토피 효과)

  • Kim, Tagon;Nho, Hwa Jung;Jun, Sang Hui;Kim, Kang Bae;Kim, Donguk
    • Korean Chemical Engineering Research
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    • v.48 no.6
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    • pp.690-694
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    • 2010
  • In this study, herbal wood vinegar including Bambusoideae, Cinnamomi Cortex, Zingiberis Rhizoma was tested to see possibility for cosmetic or skin related medicine. Anti-oxidation effect of herbal wood vinegar was tested by DPPH free radical scavenging activity, and showed 97% inhibition rate at $50{\mu}g/ml$. Anti-bacterial effect was tested by disc diffusion method, and it indicated strong anti-bacterial activity against normal skin flora Staphylococcus aureus. Whitening effect was measured by tyrosinase inhibition assay, and it was lower compared with vitamin C. Stability test was done by MTT assay, and cell toxicity was relatively high. Stability was also checked, and there was not significant change in color, aroma, appearance and pH during storage. Anti-atopic dermatitis test was done by hairless mouse and herbal wood vinegar recovered damaged skin to almost normal condition after 9 days of application. IgE concentration in herbal wood vinegar treated mouse was also reduced 30% compared with control. From the study, herbal wood vinegar showed good anti-oxidation, anti-bacterial and anti-atopic dermatitis effect, and had promising application in cosmetic or skin related medicine.

Antitumoral, Antioxidant and Antimicrobial Activities of Solvent Ftactions from Grifola umbllatus (저령추출물의 항암, 항산화 및 항균효과)

  • 하영득
    • Food Science and Preservation
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    • v.8 no.4
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    • pp.481-487
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    • 2001
  • Grifola umbellatus was extracted using methanol, and the extract was further fractionated by water and ethyl acetate. Assay of each fraction with MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium-bromide] revealed significant cytotoxicity effect of the methanol extract of Grifola umbellatus against human gastric cancer cell but not normal human lymphocytes. The methanol extract showed the highest antioxidant activity as well. Antimicrobial activity of Grifola umbellants against Helicobacter pylori was higher in method extract than in other fractions. Grifola umbellatus had a significant inhibitory effect on Helicobacter pylori reducing both its growth and urease activity. These results show that the methanol extract of Grifola umbellatus possesses therapeutic potential on gastric diseases.

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Anticancer Effect of Eriobotrya japonica Lindl by Specificity Test with Several Cancer Cell Lines (비파(枇杷)의 항암효과에 대한 암세포특이성 검정)

  • Whang, Tay-Eak;Lim, Hyun-Ock;Lee, Jae-Wa
    • Korean Journal of Medicinal Crop Science
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    • v.4 no.4
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    • pp.314-320
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    • 1996
  • Antitumouric effect of loquat was investigated treating the extract from leaves, stems, fruits or seeds to normal and cancer cell lines to check by MTT method wheather the cancer cells are spec­ifially attacked. The results are summarized as follows. Water of MeOH extract from each organ were applied to human normal and cancer cell lines, SNU-1 and SNU-C4. The water extract from fruit flesh gave no dffect to normal cell lines by killed all the cancer cell lines. The water extract from fruit flesh was purified by Sephadex LH-20 and separated into 9 fractions which were than applied to 8 cancer cell lines. The eighth fraction out of the 9 fractions gave no effect to normal cells but exerted specific cytotoxicity to breast cancer, stomach and liver cancer cells. The eighth fraction was orally administed and injected to 10 mice each suffering from the abdominal cancer induced by myeloma cells, SP2/0-Ag14. In the groups received the treatment, only one mouse each died in 2 months but the rest survived until the end of the experimental period, which those in the control plot died in 10 to 13 days. The present results confirmed that loquat contained some substance that had specific cytotoxicity to human cancer cells.

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Antioxidative, Antimutagenic, and Cytotoxic Activities of Ethanol Extracts from Cornus officianalis (산수유(Cornus officianalis) 에탄올 추출물의 항산화, 항돌연변이 활성 및 암세포 성장 억제 효과)

  • Jeon, Yeon-Hee;Kim, Mi-Hyun;Kim, Mee-Ra
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.1
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    • pp.1-7
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    • 2008
  • The antioxidative, antimutagenic and cytotoxic activities of ethanol extract from Cornus officianalis have been studied. The antioxidant activity of the ethanol extract was measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) method. The inhibition effects on the mutagenicity in Salmonella Typhimurium TA100 were evaluated by Ames test and cancer cell inhibitory effects in Hep3B cell and HeLa cell were tested by MTT assay. Cornus officianalis had an important free radical-scavenging activity towards the DPPH radical. At a concentration of 500 ppm, the DPPH radical-scavenging activity of Cornus officianalis was similar to that of L-ascorbic acid. None of the extracts produced a mutagenic effect on S. Typhimurium TA100. The ethanol extract from Cornus officianalis showed about 77% of inhibition at 500 ppm on the mutagenicity induced by 4-nitroquinoline-1-oxide. The extract from Cornus officianalis showed strong cytotoxicity against Hep3B and HeLa cells, with inhibition of 83 and 78% at a dose of $700{\mu}g$/plate, respectively. Moreover, the ethanol extracts had 34.33 mg H.E/g of polyphenols and 5.67 mg Q.E/g of flavonoids, respectively. Therefore, the present study showed antioxidative, antimutagenic and anticancer potential of the ethanol extract from Cornus officianalis.

In Vitro Study of Fluorescence Detection for Protoporphyrin IX Induced from 5-Aminolevulinic Acid in Incubated Lung Cancer Cells (폐암 세포주에서 5-Aminolevulinic Acid에 의해 유도된 Protoporphyrin IX의 형광 진단을 위한 In Vitro 연구)

  • Kim, Myung-Hwa;Kim, Hyun-Jeong;Lee, In-Seon;Kim, Kyung-Chan;Lee, Chang-Seop
    • KSBB Journal
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    • v.22 no.2
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    • pp.67-72
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    • 2007
  • This study investigates the optimal method of administrating 5-aminolevulinic acid (5-ALA) in the context of fluorescence detection by analyzing protoporphyrin IX (PpIX) fluorescence in the cultured normal and cancer cells. 5-ALA was injected as a photosensitizer to the lung cancer cells (A549, NCI-H460) and normal lung cells (HeI299). Hel299, A549, and NCI-H460 cells were incubated with various concentrations of 5-ALA ($0\sim800{\mu}g/mL$). The accumulation of PpIX induced by 5-ALA was observed in A549, NCI-H460 and Hel299 cells. The cell viability was estimated by means of the MTT assay. Formation of PpIX was measured by fluorescence spectroscopy. Especially, formation of PpIX in cancer cells was higher than normal cells. This study suggests that the difference of PpIX induced in normal and cancer cells treated with 5-ALA may use by means of fluorescence diagnosis for cancer.

Studies on Antitumor Effect and Synergistic Action of Natural Products with Anticancer drugs against Hepatic Tumors (생약의 간암세포에 대한 항종양효과와 항암제와의 상승작용)

  • Park, Gyeong-Sik;kim, Sung-Hoon;kim, Byung-tak
    • Journal of Haehwa Medicine
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    • v.4 no.1
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    • pp.211-223
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    • 1995
  • The antitumor effect of 柴胡(Bupleuri Radix : BP), 茵陳(Artemisiae capillaris Herba; ACH) 및 蒲公英(Taraxaci Herba; TH) and 蒲公英 EE層(Ethyl ether layer of TH; EETH) on human hepatocytes such as Hep G2, PLC and Hep 3B, and synergistic action with the anticancer drugs, that is, mitomycin(MMC), cisplatin(CPT) and 5-fluorouracil(5-FU) were studied by the method of MTT. The results were obtained as follows: 1. $IC_{50}$ against Hep G2, PLC and Hep 3B was $15.5{\mu}g/ml$, $25.4{\mu}g/ml$ and 31.25 in MMC, $92.5{\mu}g/ml$, $50.2{\mu}g/ml$ and $62.5{\mu}g/ml$ in CPT and $125{\mu}g/ml$ in 5-FU respectively. 2. Cytotoxic effect on Hep G2 was obvious in BP-treated group, synergistic action was most effective in TH-treated group or with MMC. 3. Cytotoxic effect on Hep 3B was obvious in ACH-treated group, synergistic action was most effective in ACH-treated group or with MMC. 4. Cytotoxic effect on PLC was obvious in ACH-treated group, synergistic action was most effective in TH-treated group or with MMC. From above results it was concluded that ACH showed the best antitumor effect against PLC and Hep 3B, BP aganst Hep G2 and also synergistic effect was most effective with MMC, which indicates that it is necessary to seperate the antitumor substances in ACH.

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산삼의 배양 및 그 응용에 관한 연구

  • 신미희
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.27 no.2
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    • pp.45-56
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    • 2001
  • Korea mountain ginseng known as oriental miracle drug is an important medicinal plant. The effect of mountain ginseng adventitious roots extract has been described. The valuable root of mountain ginseng contained several kinds of ginsenosides that have been confirmed to have many active functions for the human body. However, the study of mountain ginseng has a limit because the price of wild ginseng is very expensive and rare. The mountain ginseng adventitious roots were derived from mountain ginseng callus that were induced from mountain ginseng roots. Adventitious roots were separated from callus and grown in solid media(Murachige and stoog media). It was cultured in a 20L bioreactor. After culturing for 40days, adventitious roots were harvested. Afterwards the harvested mountain ginseng adventitious roots were dryed and extracted. We examined the effect on melanogenesis of mountain ginseng adventitious roots extract. Here, we report the inhibitory effect of melanin biosynthesis on the adventitious roots extract of In vitro test. Also, we assessed the safety of adventitious roots extract. In vitro, cytotoxicity of adventitious roots extract was assessed in mouse fibroblast using two method: The neutral red uptake assay and the MTT assay. In vivo, the allergic and irritant were Patch teated in 30 patients. Consequently, extract of mountain ginseng adventitious roots have inhibitory effect on melanin biosynthesis in B-16 melanoma cell test, tyrosinase inhibitory test and DOPA auto-oxidation test. There were decreased 86%(0.5% concentration), 45%(1% concentration) and 61%(1% concentration), respectively.

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In Vitro Study of Fluorescence Detection for Protoporphyrin IX Induced from 5-Aminolevulinic Acid in Cancerous and Normal Cells (정상 및 암 세포주에서의 5-Aminolevulinic Acid에 의해 유도된 Protoporphyrin IX의 형광 검출을 위한 In Vitro 연구)

  • Kim, Myung-Hwa;Kim, Hyun-Jeong;Lee, In-Seon;Kim, Kyung-Chan;Lee, Chang-Seop
    • KSBB Journal
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    • v.21 no.3
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    • pp.171-174
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    • 2006
  • To clarify the usefulness of fluorescent diagnosis for cancer, We investigated the optimal method of administrating 5-aminolevulinic acid(5-ALA) by analyzing fluorescence signal of Protoporphyrin IX(PpIX) in the cultured normal and cancer cells. 5-ALA was injected as a photosensitizer to the cervico-uterine cancer cell line(HeLa) and in normal liver cells(Chang). Chang and HeLa cells were incubated with various concentrations of 5-ALA($0-800{\mu}g/ml$). The accumulation of PpIX induced by 5-ALA was in HeLa and Chang cells. The cell viability was measured by MTT assay. The optimal concentration of ALA that induced maximum levels of PpIX was $50{\mu}g/ml$ in HeLa cell cultured for 24 hours after 5-ALA injection. Fluorescence of PpIX in HeLa cell was excited at a wavelength(${\lambda}$=410 nm) and showed an emission spectrum at 602.3 nm, 659.9 nm which could be related to the PpIX generation induced by the applied 5-ALA. The experimental results showed that fluorescence signal of PpIX was proportional to the concentration of 5-ALA in cancer cells, but measured with low concentration in normal cells.