• Title/Summary/Keyword: MTT reduction

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EFFECT OF METHANOL EXTRACT OF CNIDII RHIZOMA ON THE FUNCTION OF RECEPTORS FOR GABA AND GLYCINE (천궁(Cnidii Rhizoma)의 메탄올 추출물이 GABA 및 Glycine 수용체에 미치는 영향)

  • Lee, Jong-Tae;Lee, Keung-Ho;Park, Jae-Hong
    • Journal of the korean academy of Pediatric Dentistry
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    • v.32 no.1
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    • pp.55-66
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    • 2005
  • Cnidii Rhizoma (CR) was subjected to extraction with 70% methanol and tested to determine whether it has anxiolytic activity in mouse by employing staircase and rotarod tests. In addition, to understand the mechanism of anxiolytic action, CR, picrotoxin, yohimbine, isoniazid and strychnine were utilized to deliniate the potential involvement of GABA and glycine receptors in the action of Cnidii Rhizoma. To gain insights into the safety of Cnidii Rhizoma extract, behavioral and MTT tests were carried out. The results were obtained as follows: 1. CR extract had little effect on climbing numbers in the stair case test. 2. CR extract had considerable anti-anxiety effects as evidenced by the reduction of rearing numbers in the stair case test. 3. CR extract had little effect on muscle relaxation. 4. Anxiolytic actions of CR extract appeared to be mediated by glycine receptor activation. 5. Cytotoxicity in the neuronal cell was not observed and no strange behaviors were found. In short, these results indicate that CR extract has the ability to exert anxiolytic activity, possibly by activating glycine receptor with little side effects in mouse.

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The Effects of Pulsatilla Koreana for Anti - Inflammatory and Cellular Activity of Periodontal Tissue (백두옹 추출물의 치주조직 세포에 활성도 및 항염 효과에 관한 연구)

  • Jung, Jin-Gwang;Chung, Chin-Hyung;Lim, Sung-Bin;Kim, Jung-Keun;So, Eun-Hee
    • Journal of Periodontal and Implant Science
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    • v.31 no.1
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    • pp.149-165
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    • 2001
  • This study was performed to define the cytotoxicity and the anti-inflammatory action of Pulsatilla koreana extracts. To analyze cytotoxic effects, gingival and periodontal ligament fibroblasts were used, and anti-inflammatory actions related to reduction of $IL-1{\beta}$ and $PGE_2$ production were performed in vitro, for the suggestion of efficacy and safety on periodontal therapeutic use of Pulsatilla koreana extracts. We extracted ethylacetate and butylalcohol from well-dried and ground Pulsatilla koreana throughout multiple processing, then used different concentration solution(0.1 %, 0.2 %, 0.4 %, 0.01 %, 0.02 %, 0.04 %, 1 %, 2 %) of ethylacetate and butylalcohol extracts to examine eytotoxic effects and anti-inflammatory actions Cytotoxic effects were examined by ELISA reader using MTT(Methyl Thiazol-2-YL-2, 5-diphenyl Tetrazolium bromide)solution following culture of human gingival and periodontal ligament fibroblasts. Synthesis of $IL-1{\beta}$was examined by $IL-1{\beta}$ enzyme-immunoassay(EIA)system after separation and culture of monocyte, and $PGE_2$ was examined by $PGE_2EIA$ system after culture of gingival fibroblasts. The results were as follows: 1. In the MTT test of gingival fibroblasts, the change of optical density was decreased significantly at 2 % of butylalcohol extracts and 0.04 %, 0.1 %, 0.2 %, 0.4 %, 1 %, 2 % of ethylacetate extracts.(p<0.05) 2. In the MTT test of periodontal ligament cells, the change of optical density were not differ significantly. but butylalcohol and ethylacetate extracts except from butylalcohol 0.01 % showed high cell cytotoxity. 3. Both ethylacetate and butylalcohol extracts from Pulsatilla koreana inhibited the synthesis of $IL-1{\beta}$and inhibition effect of ethylacetate extracts were higher than butylalcohol extracts. 4. Both ethylacetate and butylalcohol extracts from Pulsatilla koreana inhibited the synthesis of $PGE_2$, and ethylacetate extracts were higher than butylalcohol extracts. In conclusion, ethylacetate and butylalcohol extracts from Pulsatilla koreana showed little cell cytotoxity for gingival and periodontal ligament fibroblasts, and the inhibition of $IL-1{\beta}$ and $PGE_2$ sysnthesis, therefore it is considered that these extracts can be developed as the therapeutics of the periodontal disease.

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The Anti-Cancer Effect of β-Gluconsan Calcium on Oral Cancer Cell (β-Gluconsan Calcium이 구강암 세포의 성장에 미치는 영향)

  • Kim, Ji-Hye;Jung, Yun-Sook;Kim, Hye-Young;Lee, Young-kyun;Kim, Jae-Young;Choi, Youn-Hee;Song, Keun-Bae
    • Journal of dental hygiene science
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    • v.15 no.6
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    • pp.794-799
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    • 2015
  • In recent years, there has been a global trend toward the importance of natural extracts for the prevention and treatment of human diseases. ${\beta}$-glucan is known to have anti-inflammatory activity, anti-cancer, and improvement of immune system. Polycan is purified ${\beta}$-glucan from Aureobasidum pullulans SM-2001. The anti-cancer effects of ${\beta}$-gluconsan calcium, polycan and calcium gluconate complex, were evaluated in human oral cancer YD-10B cells. YD-10B cells were cultured in the presence of 0, 0.5, 0.75, 1 mg/ml ${\beta}$-gluconsan calcium for 48 hours. MTT assay, cell counting, and observation of cell morphology were conducted. The number of cells decreased and cell morphology changed in the 0.5 mg/ml of ${\beta}$-gluconsan calcium. Almost all cells were dead in the 0.75 and 1 mg/ml. MTT assay showed a dose-dependent reduction in cell proliferation (p<0.05). These results indicate that ${\beta}$-gluconsan calcium exhibiting anti-cancer effects in YD-10B cells through changes in cell morphology and cell death.

Effect of Aceton Extract from Styela Clava on Oxidative DNA Damage and Anticancer Activity (미더덕 아세톤 추출물이 산화적 DNA 손상억제 및 암세포 독성에 미치는 영향)

  • Seo, Bo-Young;Jung, Eun-Sil;Kim, Ju-Young;Park, Hae-Ryong;Lee, Seung-Cheol;Park, Eun-Ju
    • Applied Biological Chemistry
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    • v.49 no.3
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    • pp.227-232
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    • 2006
  • Styela clava (also called as rough sea squirt or leathery tunicate) is regarded as native to the northwest Pacific region including Korea and widely distributed in parts of northwestern Europe, North America and Australia. To evaluate Styela clava as a potential bioactive agent, the antioxidant activity of aceton extracts from Styela clava (whole, substance and tunic) was tested by measuring inhibitory effect of $H_2O_2$ induced DNA damage using comet assay. Also, anticancer activity on human colon cancer cell (HT-29) was investigated by MTT reduction assay. The $200\;{\mu}M$ $H_2O_2$ induced DNA damage was inhibited with Styela clava aceton extract in dose dependent manner in human leukocytes. The maximum inhibition was by 62.8, 62.1 and 78.3% at the concentration of $50\;{\mu}g/ml$ of whole, substance and tunic extracts, respectively. The aceton extracts from S. clava were also found to inhibit the growth of human colon cancer cell. The cell proliferation rates decreased to 26.9, 30.6 and 12.0% at the concentration of $500\;{\mu}g/ml$ of whole, substance and tunic extracts, respectively. These results support that aceton extracts from S. clava may be a potential candidate as a possible antimutagenic and chemotherapeutic agent.

Neuroprotective Effects of Methanolic Extracts from Peanut Sprouts (땅콩나물 추출물의 신경세포 보호 효과)

  • Kim, Hyun-Jung;Kang, Jum-Soon;Park, Hae-Ryong;Hwang, Yong-Il
    • Journal of Life Science
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    • v.20 no.2
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    • pp.253-259
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    • 2010
  • The neuroprotective effects of extracts from various parts of peanut sprouts on glutamate-induced neurotoxicity in N18-RE-105 cells were investigated. This study was performed to evaluate the neuroprotective activity of methanolic extracts from the whole (WME), heads (HME), and stems (SME) of peanut sprouts. The neuroprotective effects of these extracts were measured by MTT reduction assay, LDH release assay, phase-contrast microscopy, and flow cytometric analysis on the N18-RE-105 cells. Among these extracts, the HME showed the greatest neuroprotective effects, and was further fractionated with hexane, diethyl ether, ethyl acetate, and water, according to degree of polarity. Out of the fractionated extracts, the diethyl ether layer showed the highest activity on glutamate-induced cytotoxicity in N18-RE-105 cells. The sub-G1 DNA contents of the glutamate-induced severely apoptotic N18-RE-105s were measured by flow cytometric analysis to confirm the HME's anti-apoptotic activity. Interestingly, after incubation with 100 mg/ml of the HME, the proportion of sub-G1 cells of the glutamate-stressed N18-RE-105s had been greatly reduced, from 58.5% to 9.1%. These results imply that HME may have strong potential as a chemotherapeutic agent against neuronal diseases.

Cytotoxic Effect of Extracts from Styela clava against Human Cancer Cell Lines (암세포주에 대한 미더덕 추출물의 세포독성 효과)

  • Jung, Eun-Sil;Kim, Ju-Young;Park, Eun-Ju;Park, Hae-Ryong;Lee, Seung-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.7
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    • pp.823-827
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    • 2006
  • The present study describes the preliminary evaluation of the cytotoxic effect from Styela clava (Korean name : miduduk) extracts. S. clava was treated with methanol, ethanol, acetone, and water, then cytotoxic effect of the extracts were evaluated by the MTT reduction assay. The ethanol extracts from S. clava showed the cytotoxic activity on the HT-29 human colon cancer cells. The ethanol extracts was further fractionated with n-hexane, diethyl ether, ethyl acetate, and water according to the degree of polarity. The diethyl ether fraction showed high cytotoxic activity of HT-29 cells. however, the other fractions showed low cytotoxicity. The diethyl ether layer also showed the cytotoxic activity against SW620, HeLa, and MCF-7 cells. These studies support that extracts from S. clava may be a potential candidate as a possible chemotherapeutic agent against human cancer cells.

Antioxidant and Neuronal Cell Protective Effect of Purple Sweet Potato Extract (자색고구마 추출물의 항산화 효과 및 신경세포 보호효과)

  • Kwak, Ji-Hyun;Choi, Gwi-Nam;Park, Ju-Hee;Kim, Ji-Hye;Jeong, Hee-Rok;Jeong, Chang-Ho;Heo, Ho-Jin
    • Journal of agriculture & life science
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    • v.44 no.2
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    • pp.57-66
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    • 2010
  • The antioxidant and neuronal cell protective effects of water extract from purple sweet potato were investigated. The total phenolics and monomeric anthocyanin contents of purple sweet potato extract were 44.25 mg/g and 2,394 mg/L, respectively. The antioxidant activities of purple sweet potato extract were evaluated using various antioxidant tests, including 1,1-diphenyl- 2-picrylhydrazyl (DPPH), 2,2'-azino- bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging activities, ferric reducing/antioxidant power (FRAP) and reducing power. In these assays, the extract of purple sweet potato presented significant radical scavenging activities, FRAP, and reducing power in a dose-dependent manner. MTT {3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl- tetrazoliumbromide} reduction assay showed significantly increase in cell viability when PC12 cells were pretreated with purple sweet potato extract. Because oxidative stress is also known to increase neuronal cell membrane breakdown, we further investigated by lactate dehydrogenase (LDH) and neutral red uptake assay. Purple sweet potato extract inhibited oxidative stress-induced membrane damage in neuronal cells. Therefore, these data results demonstrated that water extract of purple sweet potato have antioxidant activity and neuronal cell protective effect thus it has great potential as a natural source for human health.

Whitening Effect and Antioxidant Activity of Horseradish Subcritical Water Extracts (겨자무 아임계수 추출물의 항산화활성 및 미백효과)

  • In-Jae Kim;Su-Bhin Eun;Won-Hee Kim;Seon-Bhin Park;Hee-Bin Ku;Gyo-Nam Kim;Seung-Cheol Lee;Youngim Choi;Sanggeun Park;Hae-Ryong Park
    • Journal of Life Science
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    • v.34 no.4
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    • pp.236-244
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    • 2024
  • Melanin is a natural pigment found in most plants and animals, and it is involved in determining the color of the skin and hair. Melanogenesis is a reactive occurrence in melanocytes aiming to protect the skin from external stimuli, such as ultraviolet rays. Tyrosine is involved in the biosynthesis of the substrate tyrosine into melanin. However, melanin overproduction can lead to skin diseases, such as melasma, blotching, hyperpigmentation, and skin cancer. Although many studies have been conducted on whitening substances, such as kojic acid and arbutin, some countries have banned or refrained from using them due to their side effects. Therefore, this study assessed the potential of horseradish (HR) as a new whitening agent in cosmetic products. For efficient extraction, subcritical water extraction was conducted. The results showed that the horseradish subcritical water 200℃ (SW 200) extract showed high DPPH radical scavenging ability, total phenolic contents (TPC), inhibiting tyrosinase activity and inhibiting melanin production of B16-F10 melanoma cell lines. To investigate its cytotoxicity to the B16-F10 melanoma cell lines, MTT reduction assay and morphological changes were observed. No cytotoxicity was found in horseradish methanol extract and SW 200. In conclusion, this research suggests the possibility of horseradish subcritical water may be useful as a natural whitening ingredient to be used in cosmetic products.

Studies on the Anticancer Effect of Broussonetia kazinoki Extracts (닥나무(Broussonetia kazinoki) 추출물의 항암효과에 관한 연구)

  • 민경진;정승희;구성자
    • Korean journal of food and cookery science
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    • v.15 no.3
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    • pp.231-237
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    • 1999
  • The anticancer effect of the bark of Broussonetia kazinoki root extracts (hexane. chloroform, ethylacetate, butanol, aqueous) were studied. The cytotoxicity by MTT assay and inhibitory effect on the growth of sarcoma 180 cells were tested in vitro. The reduction rate of the tumor formation and spleen/body weight rate on BALB/c mouse were tested in vivo. From the tests, each fraction showed the cytotoxic effect against the sarcoma 180 cells. In addition, as the concentration of the fractions increased, cytotoxic effect tendency increased as well. The cytotoxic rate of the hexane, chloroform, ethylacetate, butanol and aqueous fractions showed by 58.7%, 40.1%, 75.7%, 52.6% and 62.7% respectively after testing by MTT assay system. And sarcoma 180 cells were incubated for 6 days at 37$^{\circ}C$ with various concentrations of each fraction. As the incubation days go on, the number of cells increased, while the inhibition rate on the growth of sarcoma 180 cells were decreased. Especially the ethylacetate fraction at the concentration of 1.0 mg/ml strongly inhibited the growth of sarcoma 180 cells by 74% compared with the control for a day 37$^{\circ}C$ The hexane, chloroform, ethylacetate, butanol and aqueous fractions inhibited on the growth of sarcoma 180 cells by 31%, 19%, 60%, 30% and 42% respectively, when sarcoma 180 cells has been incubated for 6 days at 37$^{\circ}C$. The each fraction exhibited the antitumor effect in vivo. The ethylacetate fraction reduced the tumor formation by 41% compared with the control, when sarcoma 180 cells were injected subcutaneously into the left groin of BALB/c mice. Also spleen/body weight rate of ethylacetate fraction was increased by 2.10% compared with the control (1.08%). And it is considered that there would be no toxic effect caused by each fraction of body weight and organ as there was on more changes in mouse' weight compared with the control.

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Antioxidant Activity of Glycyrrhiza uralensis Fisch Extracts on Hydrogen Peroxide-induced DNA Damage in Human Leucocytes and Cell Death in PC12 Cells

  • Lee, Hyun-Jin;Yoon, Mi-Young;Kim, Ju-Young;Kim, Yong-Seong;Park, Hae-Ryong;Park, Eun-Ju
    • Food Science and Biotechnology
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    • v.17 no.2
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    • pp.343-348
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    • 2008
  • In this study, antioxidant activity of methanol extract of Glycyrrhiza uralensis Fisch (GUE) against $H_2O_2$-induced DNA damage in human leucocytcs and cell death in PC12 cells was determined. The effect of GUE on $H_2O_2$-induced DNA damage in human leucocytcs was evaluated by the comet assay, where GUE ($1-50\;{\mu}g/mL$) was a dose dependent inhibitor of DNA damage induced by $H_2O_2$. The protective effect of GUE against $H_2O_2$-induced damage on PC12 cells was investigated by MTT reduction assay and lactate dehydrogenase release assay. A marked reduction in cell survival induced by $H_2O_2$ was significantly prevented by $1-50\;{\mu}g/mL$ of GUE. The enzyme activity of caspase-3 was elevated in $H_2O_2$-treated PC12 cells, while preincubation with GUE for 30 min inhibited $H_2O_2$-induced caspase-3 activation in a dose-dependent manner. In conclusion, GUE ameliorates $H_2O_2$-induced DNA damage in human leucocytes and has neuroprotective effect by preventing cell death in PC12 cell, suggesting that GU may be a potential candidate for novel therapeutic agents for neuronal diseases associated with oxidative stress.