• Title/Summary/Keyword: MTT assay

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Inhibitory Effect of Saengangeonbitang-gasamchilgn on Liver Fibrosis in Rat (생간건비탕가삼칠근(生肝健脾湯加三七根)이 흰쥐의 간섬유화 억제에 미치는 영향)

  • Lee, Eun;Kim, Young-Chul;Ko, Heung
    • The Journal of Internal Korean Medicine
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    • v.29 no.2
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    • pp.500-511
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    • 2008
  • The aim of this study was to investigate the inhibitory effect of Saengangeonbitang-gasamchilgn(SGGBTGSCG) on collagen production in rat hepatic stellate cells(HSC) and on the TAA-induced chronic liver injury model in rats. Methods : 1) HSCs were treated with SGGBTGSCG extract powder(50% EtOH SGGBTGSCG, dw SGGBTGSCG). After the treatment, MTT assay, BrdU assay and procollagen assay were done. In addition, gene expressions of collagen type $1{\alpha}2$, ASMA, TIMP1, and TIMP2, all of which are known to be associated with liver fibrosis, were analyzed by RT-PCR. 2) Liver fibrosis was developed in rats by injection of TAA 3 times a week for 5 weeks. After the SGGBTGSCG-treatment, body weight, liver & spleen weight, liver function test, the complete blood cell count and the change of portal pressure were studied. Results : In MTT assay, SGGBTGSCG significantly decreased the viability of HSCs in a dose-dependent manner. In BrdU assay, SGGBTGSCG significantly inhibited the HSC proliferation in a dose-dependant manner. In procollagen assay, SGGBTGSCG decreased procollagen production by HSC. In the change of rats' liver and spleen weight, TAA+SGGBTGSCG groups showed little difference compared with TAA-only group. In the liver function test, SGGBTGSCG decreased the serum level of ALT, AST, and Alp elevated by TAA. In the complete blood cell count, SGGBTGSCG significantly decreased WBC elevated by TAA and increased RBC and Hct lowered by TAA. In the change of portal pressure, SGGBTGSCG decreased portal pressure elevated by TAA. Conclusions : These results suggest that SGGBTGSCG is beneficial in the treatment of cirrhotic patients as well as for patients with chronic hepatitis.

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Efficacy of Safflower on the Acne Skin and Its Application for Facial Cleansing Biomedical Material (홍화잇꽃의 여드름피부 개선효과 및 세안용 생약식물소재 응용)

  • Park, Young-Ho;Lee, Chang-Seop
    • Journal of the Korean Chemical Society
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    • v.55 no.3
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    • pp.400-404
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    • 2011
  • Safflower is called as the 'beneficial flower' because 'it helps human health', and it was introduced as red flower in Tonguibogam due to the red color of floral leaf. From old times, it has been used for the material of cloth and rouge. Recently, polyphenol compound, the main ingredient of safflower, known as anti-aging and anti-oxidizing material in the healthy food industry becomes the emerging hot topic. This study aims to confirm by DDT (Disk Diffusion Test) assay, MTT assay, and NF-${\kappa}$B Luciferase activity inhibition assay in vitro that polyphenol compound, which is the main ingredient of safflower, has the anti-microbial efficacy to inhibit the growth of acne germs that make troubles for the teenagers or middle aged. Also it aims to evaluate its clinical efficacy on the acne skin, utilizing the facial cleansing cosmetic form of soap sample. This study can contribute to take a major step forward to the development of cosmetic soap for acne in the cosmeceutical industry.

The Effects of 5 kinds of Injinsaryung-San fractions on Cell Viability, Cell Cycle Progression and Fas-mediated Apoptosis of HepG2 Cells (인진사령산 분획물이 간세포활성, 세포주기 및 Fas-Mediated Apoptosis에 미치는 영향)

  • 고흥;이장훈;우홍정
    • The Journal of Korean Medicine
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    • v.21 no.3
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    • pp.174-185
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    • 2000
  • Objectives : This study was carried out to evaluate the effects of five fractions on cell viability, cell cycle progression and apoptosis. Methods : This study employed MTT assay, Cell cycle analysis, Cpp32 protease assay, DNA fragmentation assay and Quantitative RT-PCR analysis. Results : In MTT assay, the butanol fraction of Injinsaryung-San has showed magnificent viability, while the $H_2O$ fraction and ethylacetate fraction also showed higher viability than the control group. The $H_2O$ fraction of Injinsaryung-San has showed magnificent viability, and butanol fraction and ethylacetate fraction of Injinsaryung-San with etoposide have also showed higher viability than the only etoposide group. Cell cycle analysis showed that each fraction of Injinsaryung-San had no significant effect on the cell cycle. DNA fragmentation assay showed that the butanol fraction, $H_2O$ fraction and ethylacetate fraction carried inhibitory effects on apoptosis induction. Cpp32 protease activity assay showed that the butanol fraction, $H_2O$ fraction and ethylacetate fraction decreased Cpp32 protease activity, with the butanol fraction displaying greater effects. Quantitative RT-PCR showed that the butanol fraction, $H_2O$ fraction and ethylacetate fraction suppressed Fas and Bax genes, the butanol fraction increased BcI-2 gene, however no effect on Cpp32. Conclusions : The data shows that the butanol fraction of Injinsaryung-San increases the hepatocyte viability and has the heptocelluar protective effect by the suppression of apoptosis through gene regulation.

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In-Vitro Anticancer and Free Radical Scavenging Potential of Compound Formulation Used in Unani System of Medicine

  • Mannan, Mohd Nazir;Kazmi, Munawwar Husain;Chakraborty, Alokananda;Zakir, Mohammad;Ahmad, Tasleem;Lahari, K.
    • CELLMED
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    • v.10 no.4
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    • pp.27.1-27.6
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    • 2020
  • Cancer is one of the leading cause of mortality in India as well as worldwide. The management of cancer by conventional therapy has shown life threatening adverse effects. The researchers are now exploring the natural way of treatment. Unani system of medicine have rich literature for cancer and many compound formulations have been described in this system. Unani system of medicine is based on holistic approach and treat human being as a unit with natural herbs, mineral and animal origin drugs. An important compound Unani formulation (CUF) from the literature has been chosen to explore the Unani claim of its anticancer activity. The phytochemical constituents were assessed using standard phytochemical screening method. Antioxidant property of this formulation was assessed by DPPH assay. The DPPH free radical scavenging assay was carried out by colorimetric method and ascorbic acid was taken as a positive control. Three different extracts of CUF on different concentrations were used to screening on human breast cancer (BCC) MCF-7 cell line. For the estimation of in-vitro cytotoxic potency of the investigated extracts was assessed on MTT assay by using trypan blue method and paclitaxel was used as the standard. Hydro-ethanolic (HE) extract showed highest free radical scavenging activity among all extracts. DPPH Assay showed substantial antioxidant activity of these extracts in hydro-ethanol extract at 1㎍ concentration of CUF. The CUF showed antioxidant and anticancer activity. The claim made by Unani physician has been proved.

Protective Effect of Green Tea Extract and EGCG on Ethanol-induced Cytotoxicity and DNA Damage in NIH/3T3 and HepG2 Cells

  • Kim, Nam Yee;Kim, Hyun Pyo;Heo, Moon Young
    • Journal of Food Hygiene and Safety
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    • v.31 no.1
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    • pp.1-7
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    • 2016
  • In the present study, our aim was to determine whether green tea extract (GTE) and its major constituent, epigallocatechin-3-gallate (EGCG) have a protective effect on ethanol-induced cytotoxicity and DNA damage in NIH/3T3 and HepG2 cells. The cell viability and DNA single strand breaks were examined by MTT assay and alkaline single cell gel electrophoresis (Comet assay), respectively. Ethanol decreased the cell viability and also increased DNA single strand breaks in a concentration-dependent manner. On the other hand, GTE showed the protective effect of cytotoxicity and DNA damage induced by ethanol in both cell lines. GTE and EGCG, were found to possess the anti-oxidative and anti-genotoxic activities by evaluation with DPPH test, LDL oxidation assay, oxidative DNA damage assay and 8OH-2'dG generation test. These results were also verified by the experimental results demonstrating the lower cytotoxicity and genotoxicity of commercial green tea liqueur compared to pure ethanol in same concentration. Thus it is concluded that the supplementation of GTE or EGCG may mitigate the ethanol-induced cytotoxicity and DNA damage.

The Growth Inhibitory Effects of Epigallocatechin Gallate Against Human Skin Melanoma Cells and Human Oral Epitheloid Carcinoma Cells (Epigallocatechin gallate의 인체 피부흑색종세포와 인체 구강유상피암종세포에 대한 성장억제효과)

  • 한두석;박승택;백승화
    • Environmental Mutagens and Carcinogens
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    • v.18 no.2
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    • pp.98-103
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    • 1998
  • Epigallocatechin gallate (EGCG) was reported to exert weak cytotoxicity against normal healthy cells such as C3H10T1/2 cells, but profound inhibitory effects on the initiation or promotion stage of chemical carcinogenesis in mammary gland, blood and mouse skin. This study was carried out to develop antitumor agents with weak side effects and strong antitumor activity. Human skin melanoma cells (HBT 69) and human oral epitheloid carcinoma cells (OCL 17) were cultured in RPMI-1640 media containing 10% fetal bovine serum, antibiotic, and fungizone. After incubation for 24 hrs, the cells were treated with various amounts of (EGCG) for 48 hrs. The growth inhibitory effects of EGCG in human oral epitheloid carcinoma cells were evaluated by the 3- (4,5-djmethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), neutral red (NR), and sulforhodamine B protein (SRB) assays of colorimetric methods. The light microscopic study was also carried out to observe morphological changes of the treated cells. These results obtained were as follows; 1. Significantly inhibitory effects of EGCG against cultured human oral epithelioid carcinoma cells. 2. Significantly inhibitory effects against cultured human skin melanoma cells treated with 50 $\mu$M EGCG, but decreased inhibitory effects in 100 $\mu$M EGCG. 3. Degenerative changes against cultured human oral epitheloid carcinoma cells. 4. Degenerative changes against human skin melanoma cells treated with 50 UM EGCG, but recovered degenerative changes in 100 $\mu$M EGCG.

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Antiviral Effects of Fermented Lonicerae Flos on A Type Influenza Virus (발효 금은화의 인플루엔자 바이러스 A형에 대한 저해효과)

  • Suhr, Sung-Sook;Jung, Sung-Ki
    • The Journal of Internal Korean Medicine
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    • v.30 no.3
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    • pp.465-480
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    • 2009
  • Objective : Lonicerae Flos has detoxifying properties and been used as antipyretic, antibacterial and antitumor. Fermentation of herbal medicine is known to increase the absorption, enhance effectiveness, decrease herbal toxicity and reduce side-effects. This study was performed to measure the effects of fermented Lonicerae Flos on influenza A/WSN (H1N1) virus replication. Material and Methods : Lonicerae Flos was fermented by Lactobacillus casei PM1. Fermented Lonicerae Flos was treated for 12 hours to MDCK (Mardin Darby canine kidney) cells, then cell-virulence was observed by MTT assay for 12 hours, 24 hours, and 36 hours after treatment. Following cases were conducted for 0, 10, 100, and $1000{\mu}g/ml$ concentrations of fermented Lonicerae Flos under the same time-frame; the fermented Lonicerae Flos was treated to MDCK cells before and after contamination by A-type influenza virus. The fermented Lonicerae Flos and the virus were mixed directly. The influence was observed by MTT assay and plaque assay. Results : These findings suggest that the fermented Lonicerae Flos inhibited the virulence of influenza A virus in MDCK cells and suppressed the plaque forming colonies induced by influenza A virus. Furthermore, pretreatment with fermented Lonicerae Flos was more effective than post-treatment. The titer of influenza virus was reduced for all before and after influenza A virus inoculation.

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Antimutagenic and Antileukemic Activities Aloe vera L.

  • Lee, Keyong-Ho;Kang, Hee-Gon;Cho, Choa-Hyoung;Lee, Moon-Joon;Lee, Jae-Hyun;Kim, Chang-Han
    • Natural Product Sciences
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    • v.6 no.2
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    • pp.56-60
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    • 2000
  • We investigated that the extract of Aloe vera L. and its fractions exert antimutagenic activity against Salmonella typhimurium TA98 and TA100, and antileukemic effect against K562 human leukemia cell line. The aqueous ethanolic extract of A. vera L. was revealed to have antimutagenic effect on the AF-2 (2-(2-furyl)-3-(5-nitro-2-furyl)-acrylamide) in Salmonella mutation assay. Among the three fractions (fractions A, B and C) separated by silica gel chromatography, fraction C $(50\;{\mu}g/plate)$ exhibited the greatest antimutagenic effect on the AF-2 with inhibition rate of 84 and 90% in Salmonella typhimurium TA98 and TA100, respectively. The fraction C $(500\;{\mu}g/ml)$ inhibited the growth of K562 human leukemia cell line by 93% in MTT assay. However, the components of A. vera L. did not exhibit cytotoxic effect against MDBK bovine normal kidney in MTT assay.

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Selective Cytotoxicities of Phenolic Acids in Cancer Cells (페놀산의 구조가 암세포에 대한 세포독성에 미치는 영향)

  • 한두석;오상걸;오은상
    • Toxicological Research
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    • v.19 no.1
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    • pp.45-50
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    • 2003
  • The purpose of this study was to determine the role of substituted groups in phenolic compounds to develop an anticancer agent having strong cytotoxicity against cancer cells but weak against normal cells. The phenolic compounds used in this study were gallic acid and ferulic acid with hydroxyl and carboxyl groups, syringic acid with hydroxyl, carboxyl and methoxy groups, and pyre-gallol with hydroxyl groups. Cytotoxicities of these compounds were evaluated by MTT assay for cell viability and XTT assay for cell adhesion activity in normal human skin fibroblast (Detroit 551) and human skin melanoma (SK-MEL-3) cells. Syringic acid, gallic acid and ferulic acid decreased the cell viability and cell adhesion activity in SK-MEL-3 cells but not in Detroit 551 cells while pyrogallol decreased in both cells. The susceptibility of cell viability based on the $IC_{50}$ values of MTT assay in Detroit 551 cells was in the following order: pyrogallol > gallic acid > ferulic acid > syringic acid, while it was in SK-MEL-3 cells: Syringic acid > progallol > ferulic acid > gallic acid. These results suggest that carboxyl and methoxy groups of these compounds play an important role in selectivity of cytotoxicity in normal and cancer cells.

Cytotoxicity of Folkloric Medicine in Murine and Human Cancer Cells (천연물로부터 항암물질의 분리)

  • Lee, Ihn-Rhan;Song, Ji-Young;Lee, Yun-Sl
    • Korean Journal of Pharmacognosy
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    • v.23 no.3
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    • pp.132-136
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    • 1992
  • The whole plants of Selaginella tamariscina, Orostachycis japonicus, the cortex of Ulmus mandshurica, and the wood of Alnus japonica have been used as folk medicine for treating cancer. The cytotoxic activity of these plants were tested using a calorimetric tetrazolium assay (MTT assay). S. tamariscina and A. japonica showed mild $IC_{50}$ value, comparing with O. japonicus and U. mandshurica. So, MeOH extracts of S. tamariscina and A. japonica were partitioned into $CHCl_3$, EtOAc and n-BuOH, successively. The $CHCl_3$, EtOAc and BuOH fractions of S. tamariscina and A. japonica showed low percent of survival against $P_{388}$ and $MKN_{45}$ cells respectively. To isolate active components, they were subjected to silica gel column chromatography. Compound I was obtained from EtOAc extracts of S. tamariscina and identified as amentoflavone by chemical and spectral analysis. Amentoflavone inhibited the survival of P388 cells dose dependently, while not clearly inhibited that of $MKN_{45}$ cells.

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