• Title/Summary/Keyword: MTT analysis

Search Result 746, Processing Time 0.028 seconds

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
    • /
    • v.21 no.3
    • /
    • pp.174-185
    • /
    • 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.

  • PDF

Design Optimization of a Paper Feeding Mechanism using Numerical Analysis Program (수치해석 프로그램을 이용한 미디어 이송 장치의 기구학적 최적설계)

  • Lee S.G.;Choi J.H.;Bae D.S.;Cho H.J.;Song I.H.;Kim M.S.
    • Proceedings of the Korean Society of Precision Engineering Conference
    • /
    • 2006.05a
    • /
    • pp.107-108
    • /
    • 2006
  • This paper shows the design optimization of the paper feeding mechanism under dynamic behavior by using commercial codes of RecurDyn/MTT2D and RecurDyn/AutoDesign which are developed by functionBay, Inc. A virtual mockup for dynamics analysis of the paper feeding mechanism is build on RecurDyn/MTT2D and is simulated. Flexible paper is represented as a series of rigid bars connected by revolute joints and rotational spring dampers. Paper is fed by a contact and friction mechanism on rollers or guides. The slip of the paper and nip force of rollers are measured to estimate the system performance. After a simulation, these performances are automatically send to RecurDyn/AutoDesign which is a sequential approximate optimization tool based on the response surface modeling. RecurDyn/AutoDesign makes the approximate objective function and computes the optimized design points of the design variables and gives them to analysis tool. And then the simulation is repeated with the updated design variables. These processes are repeated until finding a tolerable design optimization. In this paper, a paper feeding mechanism is introduced and it is optimized with the proposed algorithms.

  • PDF

AN EXPERIMENTAL STUDY OF THE DEGREE OF CONVERSION AND CYTOTOXICITY OF DUAL CURE RESIN CEMENTS (수종 이원중합 레진 씨멘트의 중합률 및 세포 독성에 관한 실험적 연구)

  • Roh, Byoung-Duck;Park, Seong-Ho;Lee, Chung-Suck
    • Restorative Dentistry and Endodontics
    • /
    • v.20 no.1
    • /
    • pp.33-54
    • /
    • 1995
  • The degree of conversion of composite resin was known to have influence on the mechanical properties of composite materials such as hardness, strength, wear resisitance, dimensional and color stability. Also unreacted monomer was reported to be harmful to the pulp. So the degree of conversion was a very important factor in the success of composite resin restorations. In recent, the dual cure resin cement was developed with the advocations that it could increase the curing rates in the sites where the curing ligt could not reach. Moreover many manufactors added some adhesive components in the resin cement. This study was undertaken to observe the effects of curing depth and light curing times on the degree of conversion of dual cure resin cements. CR INLAY CEMENT, DUAL CEMENT and OPTEC BOND, by the Fourier transform Infrared analysis, changing the curing depth 1mm, 2mm and 3mm, and varying the light curing time 20 seconds, 40 seconds and 80 seconds at each depth. The cytotoxicity of dual cure resin cements was tested by the in vitro MTT method using L929 cell. The results was evaluated and compared statistically. The results were obtained as follows : 1. The dual cure resin cements reavealed various degree of conversion, CR INLAY CEMENT and DUAL CEMENT had a tendency to be more reactive to the light cure and OPTEC BOND was a more chemical one. 2. CR INLAY CEMENT and DUAL CEMENT showed the lowest degree of conversion in 2 mm depth, and in 3mm depth the degree of conversion increased, which were due to the chemical cure of dual cures, but OPTEC BOND showed decreasing degree of conversion with increasing curing dept h and all experimental groups showed lower degree of conversion than CHEMICAL group which cured in dark room with no light, so the weak light-curing of dual cure resin cement prevented the chemical cure. (P<0.05) 3. CR INLAY CEMENT and DUAL CEMENT showed increasing degree of conversion in 1 mm and 3 mm, according to the increasing cure times, but in 2 mm depth the degree of conversion decreased with increasing light-curing times and OPTEC BOND showed contrary tendency, but there was no ststistical importance in the differences among the experimental group.(P>0.05) 4. The optical density by MTT assay of extractions of CR INLAY CEMENT, DUAL CEMENT and OPTEC BOND revealed no statitically important differences comparing with optical density of negative control.(P>0.05) 5. CR INLAY CEMENT showed a tendency of increaing cytotoxicity with days and DUAL CEMENT and OPTEC BOND showed higher cytotoxicity in 2 days than in 4 days, but there was no statistical importance in the differences.(P>0.05).

  • PDF

Anti-inflammation and Anti-aging Effect of Lactic Acid Bacteria Fermented Lycium chinense Extracts (젖산균 발효 구기자(Lycium chinense ) 추출물의 항염증 및 항노화 효능 연구)

  • Han, Dong-Geun;Lee, Ju-Seong;Gal, Gyeok;Park, Nam-Wan;Kim, Hyun-Jeong;An, Bong-Jeon
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • v.48 no.2
    • /
    • pp.113-121
    • /
    • 2022
  • In this study, fermented Lycium chinense (L. chinense) extract was used to analyze the efficacy of nitric oxide (NO) and anti-aging inhibition to verify its usefulness as a functional cosmetic material. L. chinense was extracted with hydrothermal and 70% ethanol as a solvent, fermented, concentrated, and freeze-dried to prepare a sample, followed by MTT assay, NO inhibition assay, western blot assay, and UPLC analysis.As a result of the analysis of NO inhibitory efficacy, fermented L. chinense water extract (FLW) and fermented L. chinense 70% ethanol extract (FLE) showed excellent NO inhibitory efficacy of 47.96% and 56.71%, respectively. As a result of measuring the expression patterns of the wrinkle-related proteins MMP-1, and TRPV-1 through Western blot, both FLW and FLE confirmed the inhibitory efficacy of MMP-1 and TRPV-1. Based on the results of the experiment, the fermented L. chinense extract is expected to have a high application value as a functional cosmetic material related anti-inflammatory, and anti-aging.

Paclitaxel Induced Caspase-Independent Mitotic Catastrophe in Rabbit Articular Chondrocyte (Paclitaxel에 의한 관절연골 세포의 capase-비의존적 mitotic catastrophe 유도)

  • Im, Jeong-Hee;Kim, Song-Ja
    • Journal of Life Science
    • /
    • v.20 no.4
    • /
    • pp.519-527
    • /
    • 2010
  • Paclitaxel is known as a potent inhibitor of microtubule depolymerization. It leads to mitotic arrest and cell death by stabilizing the spindle in various cell types. Here, we investigated the effects of paclitaxel on the proliferation and cell death of rabbit articular chondrocytes. Paclitaxel inhibited proliferation in a dose- and time- dependent manner, determined by MTT assay in rabbit articular chondrocytes. We also established paclitaxel-induced G2/M arrest by fluorescent activated cell sorter (FACS) analysis. Paclitaxel increased expression of cyclin B, p53 and p21, while reducing expression of cdc2 and cdc25C in chondrocytes, as detected by Western blot analysis. Interestingly, paclitaxel showed the mitotic catastrophe that leads to abnormal nucleus division and cell death without DNA fragmentation through activation of caspase. Cell death by mitotic catastrophe in cells treated with paclitaxel was suppressed by inhibiting G1/S arrest with 2 mM thymidine. These results demonstrate that paclitaxel induces cell death via mitotic catastrophe without activation of casepase in rabbit articular chondrocytes.

Effects of Sophorae Radix on Human Colorectal Adenocarcinoma Cells (고삼의 인체 대장암세포에 미치는 효과)

  • Kim, Min-Chul;Lee, Hee-Jung;Lim, Bo-Ra;Kim, Hyung-Woo;Kim, Byung-Joo
    • Journal of Physiology & Pathology in Korean Medicine
    • /
    • v.26 no.2
    • /
    • pp.155-159
    • /
    • 2012
  • The purpose of this study was to investigate the anti-cancer effects of Sophorae Radix and the effects of 5-Fluorouracil (5-FU) in human colorectal adenocarcinoma cells (HT-29). We used human colorectal adenocarcinoma cell line, HT-29 cells. We examined cell death by MTT assay and caspase 3 assay with Sophorae Radix. To examine the inhibitory effects of Sophorae Radix, cell cycle (sub G1) analysis was done the HT-29 cells after three days with Sophorae Radix. The reversibility of Sophorae Radix was examined on one day to five days treatment with $150{\mu}g$ Sophorae Radix. Sophorae Radix inhibited the growth of HT-29 cells in a dose-dependent fashion. Also we showed that Sophorae Radix induced apoptosis in HT-29 cells by MTT assay, caspase 3 assay and sub-G1 analysis. Sophorae Radix combined with 5-FU markedly inhibited the growth of HT-29 cells compared to Sophorae Radix or 5-FU alone. After 3 days treatment of HT-29 cells with Sophorae Radix, the fraction of cells in sub-G1 phase was much higher than that of the control group. Our findings provide insight into unraveling the effects of Sophorae Radix in human colorectal adenocarcinoma cells and developing therapeutic agents against colorectal cancer.

Anti-cancer Effects of Scutellaria barbata in AGS Human Gastric Adenocarcinoma Cells (인체 위암세포주에서 반지련(半枝蓮)의 항암 효능에 관한 연구)

  • Shim, Ji Hwan;Lee, Soojin;Gim, Huijin;Park, Hyun Soo;Kim, Byung Joo
    • Journal of Physiology & Pathology in Korean Medicine
    • /
    • v.29 no.2
    • /
    • pp.195-201
    • /
    • 2015
  • The aim of the study is to investigate the anti-cancer effects of Scutellaria barbata in AGS human gastric adenocarcinoma cells. MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) assay and caspase 3 or 9 activity assay were carried out to examine cell death with Scutellaria barbata. To elucidate the inhibitory effects of Scutellaria barbata, cell cycle (sub-G1) analysis and mitochondrial membrane potential were performed in AGS cells after 24 h incubation with Scutellaria barbata. Scutellaria barbata induced apoptosis in AGS cells by using the MTT assay, the sub-G1 analysis and mitochondrial membrane potential assay. The stronger inhibition effects of AGS cell growth was observed by application of Scutellaria barbata combined with several anti-cancer drugs (paclitaxel, 5-fluorouracil, cisplatin, ectoposide, doxorubicin and docetaxel) in comparison to the application of Scutellaria barbata or anti-cancer drugs. Our findings provide insight into unraveling the effects of Scutellaria barbata in human gastric cancer cells and developing therapeutic agents against gastric cancer.

The Effect of Bee Venom & Melittin Solution on Cell Death in Synovial Cell Line (봉독(蜂毒) 및 Melittin 약침액(藥鍼液)이 관절염(關節炎) 활액세포(滑液細胞)에 미치는 영향(影響))

  • Han, Sang-won;Park, Ki-hyeon;Jung, Tae-young;Seo, Jung-chul
    • Journal of Acupuncture Research
    • /
    • v.19 no.4
    • /
    • pp.74-88
    • /
    • 2002
  • Objectives : This study is aimed to investigate the effects of bee venom and melittin on cell death in synovial cell line. Methods : It was evaluated by using MTT assay, morphologic method, DNA fragmenation, NO generation, flow cytometry, immunocytochemistry analysis, RT-PCR, Western blot. Results : The obtained results are summarized as follows: 1. The MTT assay demonstrated that synovial cell viability was significantly inhibitted dose-dependently by treatment with bee venom and melittin in comparison with control. 2. The morphologic study demonstrated that synovial cell showed apoptosis after treatment with bee venom and melittin for 6 hours using microscope. 3. In case of NO generation bee venom group and melittin group showed significant inhibition in comparison with control. 4. The Flow cytometry demonstrated that apoptosis of synovial cell treated with bee venom and melittin was related with stop of cell cycle in stage of $G_0/G_1$. 5. DNA fragmenation demonstrated that synovial cell treated with bee venom and melittin showed DNA ladder below l Kbp. 6. Immunocytochemistry assay demonstrated that COX-II and PLA2 were strongly down-regulated by treatment with bee venom and melittin whereas iNOS was almostly not expressed by bee venom treatment and slightly expressed by melittin treatment. 7. RT-PCR analysis demonstrated that iNOS were strongly down-regulated by treatment with bee venom and melittin whereas COX-II was almostly not expressed by bee venom treatment and slightly expressed by melittin treatment. 8. Western blot demonstrated that iNOS were strongly down-regulated by treatment with $15{\mu}g/ml$ bee venom whereas COX-II was strongly down-regulated from $5{\mu}g/ml$ bee venom. Conclusions : These results suggest that bee venom and melittin have significant effect on cell death in synovial cell line and further study is needed in vivo.

  • PDF

Anti-inflammatory Effect of LFR on LPS-stimulated THP-1 Cells

  • Sul, Mu-Chang;Kim, Hong-Yeoul;Roh, Young-Lae;Jung, Hee-Jae;Jung, Sung-Ki
    • The Journal of Internal Korean Medicine
    • /
    • v.30 no.2
    • /
    • pp.388-398
    • /
    • 2009
  • Background and Objective: Luffae Fructus Retinervus (LFR) is used for investigating symptoms of inflammation. We have evaluated the anti-inflammatory effect of LFR by analyzing the expression of pro-inflammatory cytokines. Materials and Methods : We differentiated THP-l cells into macrophage-like cells by treatment with PMA. Inflammation was induced by treatment with LPS and PMA. We determined the safe concentration of LFR by using the MTS and MTT assays and using PD 98059 as a negative control for comparison of the anti-inflammatory effect of LFR. Results : The MTS and MTT analysis showed that the cell survival rate was >80% within the LFR concentration range of 10-100 ng/ml and began to decrease to >80% at 1 ${\mu}g/ml$. By RT-PCR analysis, the gene expression of TNF-${\alpha}$, IL-8, TGF-${\beta}$, IL-6, IL-${\beta}$1, and IL-10 levels were down-regulated when monocyte-derived macrophages were treated with concentrations of LFR between 10 ng/mL and 100 ng/mL. Conclusion : We conclude that LFR exerts an anti-inflammatory effect by inhibiting the expression of pro-inflammatory activity. The results suggest a promising way to treat general inflammatory diseases.

  • PDF

Effects of the Bee Venom on Human Gastric Adenocarcinoma Cell Lines (봉독이 위암 세포주에 미치는 효과)

  • Heo, Gyeong;Kim, Myung Ho;Lim, Seong Woo
    • Journal of Physiology & Pathology in Korean Medicine
    • /
    • v.27 no.1
    • /
    • pp.92-98
    • /
    • 2013
  • Bee Venom(below BV) has been used in alternative medicine to treat the diseases, such as pain diseases. BV contains a variety of peptides, including melittin, apamin, adolapin, MCD peptide, enzymes(i.e. PLA2), amines(i.e. histamine and epinephrine), and nonpeptide components. The two main components of BV are melittin and PLA2. The cell cytotoxic effects through the activation of PLA2 by melittin have been suggested to be the critical mechanism for the depress of cancer cell. Melittin and PLA2 have been reported to induce apoptosis and to possess anti-cancer effects and neurite outgrowth in PC12 cells. Analysis of proliferation was confirmed by MTT assay. BV decreased cell number through dose- and duration-dependent manner and these effects are apparent at a concentration of 3 ${\mu}g/ml$. To observe which signaling molecules will be activated by BV, phosphorylation of ERK, p38 MAPK, JNK and ERM were examined by Western blot analysis. To study the long term effect of BV in human gastric adenocarcinoma cell lines, the image of cells treated with BV for 4 days were obtained. BV was shown to exhibit anti-cancer activity in human gastric adenocarcinoma cell lines at a broad range of concentrations of 3 ${\mu}g/ml$. ERK, p38 MAPK and JNK were found to increase in BV treated cells. However, ERM which known to be involved in the cell death, was gradually decreased to 30minutes after addition 3 ${\mu}g/ml$ of BV. These results provide a possible BV-induced inhibitory signal for cancer proliferation that is initiated by the decrease in ERM activity. Moreover, it is likely that the activation of ERK, p38 MAPK and JNK are required for the BV-induced inhibition of cancer proliferation.