• Title/Summary/Keyword: MTT method.

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The Effects of Antioxidant and Anti-Alzheimer on Hydrogen peroxide and $\beta$-amyloid peptid-induced PC 12 cells by Semen Ziziphi Spinosae water extract ($H_{2}O_2$와 A$\beta$로 유도된 pc12 cell에서 생산조인(生酸棗仁) 수추출물의 항산화 및 항치매 효과)

  • Lee, Sang-Won;Kim, Dae-Hyun;Yun, Jong-Hyun;Kim, Jin-Woo;Jung, Ejun-Young;Lee, Seoung-Geun;Lee, Key-Sang;Kim, Tae-Heon;Lyu, Yeoung-Su;Kang, Hyung-Won
    • Journal of Oriental Neuropsychiatry
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    • v.19 no.3
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    • pp.179-193
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    • 2008
  • Objective: The antioxidant and anti-Alzheimer effects of Semen Ziziphi Spinosae (SZS) water extract against the amyloid beta peptide (1-42) or H202-induced oxidative damage and cell death were investigated in rat pheochromocytoma line PC 12. Methods: The cells were incubated with SZS water extract and oxidative damage-inducing materials, amyloid beta peptide (1-42) or H2O2 for 24 h. The cellular viability was assessed by WST-1 assay, cytotoxic damage by LDH activity assay, oxidative damages of cells by fluorescence spectrophotometric method, and apoptosis by TUNEL staining assay. Results and Conclusions: 1. Preincubation of the cells with SZS water extract prior to amyloid beta peptide (1-42) (2 uM) or H2O2 (30 uM) exposure elevated the cell survival close to the control and decreased the level of LDH activity and the fluorescence from the cell homogenates and TUNEL staining of the cells, compared to only amyloid beta peptide (1-42) (2 uM) or H2O2 (30 uM) treated conditions. 2. Our study suggests that Semen Ziziphi Spinosae (SZS) water extract has protective effects against amyloid beta peptide (1-42) or H2O2-induced cell toxicity through the antioxidation mechanism, which might be beneficial for the treatment of Alzheimer's disease.

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The Effect of the Compound of Tomato Extract to the Prostatic Cancer Cell and the Prostate of the Rat Model of Benign Prostatic Hyperplasia (토마토 추출액 복합체가 전립선 암 세포와 전립선 비대증에 미치는 영향)

  • Kang, Han-Saem;Kim, Gwang-Yun;Jung, Il;Oh, Sung-Dug;Kim, Chang-Hoon;Shim, Bong-Sup;Park, Keun-Hyung;Oh, Suk-Jung
    • Korean Journal of Pharmacognosy
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    • v.38 no.2 s.149
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    • pp.197-203
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    • 2007
  • Benign prostatic hyperplasia (BPH) is one of the common disease in elderly men. Recently old-age population is increased and we are growing more and more interested in clinical importance of BPH. In this study, the effect of PLX, which was the mixture of tomato extract (including 2% of lycopene) and chitooligosaccharide, on prostatic cancer cell and testosterone-induced BPH in adult rats of the Sprague Dawley strain was determined. The cell viability was evaluated by MTT method using L929 and LNCaP cell line, pretreated with various concentrations of PLX. The expression of prostatic specific antigen (PSA) and 5${\alpha$}$-reductase genes were evaluated by realtime PCR using LNCaP cell line and compared various concentrations of PLX with 50 ${\mu}$M of finasteride. An experimental prostatic hyperplasia was induced in male Sprague Dawley rats by giving testosterone for 8 weeks. After 2 weeks from start of giving testosterone, PLX and finasteride were administered orally once a day. The results were analyzed with prostate weight per body weight at 8 weeks. Cell viability of L929 cell line decreased specifically at the concentration of 2000 ${\mu}$g/mf of PLX. The cytotoxicity of PLX to the LNCaP cell line was shown at above 500 ${\mu}$g/ml of PLX. The inhibitory effect of PLX to the expression of PSA and 5${\alpha$}$-reductase genes in LNCaP cell line increased with the concentration of PLX. In vivo study, the results of PLX and finasteride administered group were 3.75${\pm}$0.60 and 3.49${\pm}$0.49 prostate weight ${\times}10^3$/body weight, which were lower than the result of BPH induced group (4.74${\pm}$0.58). These results suggested that PLX may be an effective material in BPH by having the role of the 5a-reductase inhibitor.

Anti-inflammatory Effects of Wiryeong-tang Mix Soft Extract (위령탕 혼합단미연조엑스의 항염증 효과)

  • Kim, Myoung-Jin;Kim, Se-Jin;Nam, Won-Hee;Leem, Hyun-Hee;Jeon, Young-Hee;Son, Su-Mi;Kim, Jung-Ok
    • Herbal Formula Science
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    • v.28 no.4
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    • pp.339-349
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    • 2020
  • Objectives : Wiryeong-tang (WRT) is a traditional herbal medicine used to treat kidney-related diseases. However, the anti-inflammatory and anti-gastritis effect of Wiryeong-tang was not well known. Therefore, we experimented to confirmed the anti-inflammatory and anti-gastritis effects of Wiryeong-tang. Methods : The RAW 264.7 cells were pre treated with Wiryeong-tang mix soft extract (WRT-mse; 50, 100 and 200 ㎍/mL) for 1 hrs, and then incubated with lipopolysaccharide (LPS; 500 ng/mL). Cell viability was measured by the MTT method, and nitric oxide (NO) was measured with griess reagent. In addition, pro-inflammatory cytokines were measured by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). For anti-gastritis effect in vivo, acute gastritis was induced using 150 mM HCl/60% ethanol used ICR mice. WRT-mse (133 mg/kg) was pre treated for 3 days and then treated with 150 mM HCl/60% ethanol 1 hrs later. Then gastritis was observed and inflammatory cytokines in the gastric tissue was measured. Results : The 8 marker components of the WRT-mse were determined by simultaneous analysis using HPLC. WRT-mse was not toxic and inhibited pro-inflammatory cytokines such as IL-1β, IL-6 and TNF-α at NO production, protein and mRNA levels. Also, it was confirmed that WRT-mse improved bleeding and edema in gastritis, and suppresses inflammatory cytokines. Conclusion : In summary, our results suggest that the treatment of the WRT-mse reduced and improved the 150 mM HCl/60% ethanol induced acute gastritis and the inflammation caused by LPS stimulation in RAW 264.7 cells. Therefore, this study may provide useful drug or clinical evidence for WRT-mse to prevent inflammation.

The Anticancer Mechanisms of Taxol-Diethylenetriamine pentaacetate Conjugate in HT29 Human Colorectal Cancer cells

  • Lee, Na-Kyung;Kim, Hyun-Jeong;Yang, Seung-Ju;Kim, Yoon-Suk;Choi, Hyun-Il;Shim, Moon-Jeong;Awh, Ok-Doo;Kim, Tae-Ue
    • BMB Reports
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    • v.34 no.3
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    • pp.237-243
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    • 2001
  • Taxol, a natural product extracted from the Taxus brevifolia, is known to have significant anti-tumor activities against many common cancers, including ovarian and breast cancers. Despite the pronounced anti-tumor activity of this compound, its poor solubility in aqueous solutions hampers its clinical applications. We studied the anticancer mechanisms of the water-soluble taxol diethylenetriamine pentaacetate (DTPA) used for radiolabeling, and compared it to that of taxol. In vitro cytotoxicities of taxol and taxol-DTPA conjugate were tested in HT29 human colorectal cancer cells by the MTT method. As the result, the $IC_{50}$ value of the taxol-DTPA conjugate was about three fold higher than that of taxol. When analyzed by an agarose gel electrophoresis, the DNA ladders became evident after the incubation of cells with the taxol-DTPA conjugate for 24 h. We also found morphological changes of the cells undergoing apoptosis with electron microscopy Next, we examined the signal pathway of taxol-DTPA conjugate-induced apoptosis in HT29 cells. The activation of extracellular signal-regulated protein kinase (ERK1/2) occurred at 10, 30, 60 and 120 min after 200 nM taxol-DTPA conjugate treatment. The pretreatment of the MEK inhibitor (PD98059) completely blocked the taxol-DTPA conjugate-induced ERK1/2 activation. The activated ERK1/2 translocated into the nucleus at the same time and phosphorylated its transcriptional factor, c-Jun. These results suggest that the taxol-DTPA conjugate has an apoptotic activity in HT29 cells, and that its proapoptic activity might be related with the signal transduction via ERK1/2 and c-Jun similar to that of taxol.

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Evaluation of Antioxidant Activity of Sugar Alcohols Using TOSC (Total Oxy-radical Scavenging Capacity) Assay (TOSC 법을 이용한 당알코올의 항산화 활성 평가)

  • Kang, Keon-Wook;Kwak, Sang-Hoon;Yun, Sei-Young;Kim, Sang-Kyum
    • Toxicological Research
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    • v.23 no.2
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    • pp.143-150
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    • 2007
  • Although animal and epidemiological studies have suggested oxidative stress as an etiological factor in pathogenesis including cancer, inflammation, sepsis, fibrosis, cardiovascularlneurodegenerative diseases and aging-related disorders, conflicting results have been obtained in clinical trial with antioxidants. The reason for this discrepancy remains unknown but may be due, in part, to the lack of a validated assay system for evaluating antioxidant capacity. The antioxidant activity of a series of sugar alcohols against peroxyl radicals, hydroxyl radicals and peroxynitrites was determined by the total oxy-radical scavenging capacity (TOSC) assay and cell-based assay using H4IIE cells. Specific TOSC values calculated from the slope of the linear regression for erythritol, xylitol, sorbitol or mannitol against peroxyl radicals was $2.1{\pm}0.2,\;3.7{\pm}0.3,\;9.1{\pm}0.3$ or $8.7{\pm}1.1$ TOSC/mM, respectively. Specific TOSC values for erythritol, xylitol, sorbitol or mannitol against peroxynitrite was $1.9{\pm}0.3,\;3.9{\pm}0.4,\;7.8{\pm}0.7$ or $7.7{\pm}0.5$ TOSC/mM, respectively. These results suggest that oxy-radical scavenging capacity is dependent on the number of aliphatic hydroxyl group in sugar alcohols of monosaccharide. Tert-butylhydroperoxide (t-BHP)-induced cell toxicity determined by MTT assay was marginally attenuated by 10 mM erythritol, but completely inhibited by 10 mM xylitol, 2 mM sorbitol or 0.75 mM maltitol, a disaccharide alcohol. Oxidative stress markers, such as glutathione (GSH) and malondial-dehyde (MDA) levels, were measured in t-BHP-treated cells using HPLC equipped with a fluorescence detector and a reverse phase column. Erythritol did not change the levels of GSH and MDA in H411E cells treated with t-BHP. The t-BHP-induced changes in cellular GSH and MDA levels were ameliorated by 10 mM xylitol and completely blocked by 10 mM sorbitol and maltitol. These results indicate that sugar alcohols protect cells against oxidative stress via scavenging oxy-radical and suggest that TOSC assay in conjunction with cell-based assay is a valid method for evaluating antioxidant capacity of natural and synthetic chemicals.

Anti-tumor Effects of Penfluridol through Dysregulation of Cholesterol Homeostasis

  • Wu, Lu;Liu, Yan-Yang;Li, Zhi-Xi;Zhao, Qian;Wang, Xia;Yu, Yang;Wang, Yu-Yi;Wang, Yi-Qin;Luo, Feng
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.489-494
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    • 2014
  • Background: Psychiatric patients appear to be at lower risk of cancer. Some antipsychotic drugs might have inhibitory effects on tumor growth, including penfluridol, a strong agent. To test this, we conducted a study to determine whether penfluridol exerts cytotoxic effects on tumor cells and, if so, to explore its anti-tumor mechanisms. Methods: Growth inhibition of mouse cancer cell lines by penfluridol was determined using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Cytotoxic activity was determined by clonogenic cell survival and trypan blue assays. Animal tumor models of these cancer cells were established and to evaluate penfluridol for its anti-tumor efficacy in vivo. Unesterified cholesterol in cancer cells was examined by filipin staining. Serum total cholesterol and tumor total cholesterol were detected using the cholesterol oxidase/p-aminophenazone (CHOD-PAP) method. Results: Penfluridol inhibited the proliferation of B16 melanoma (B16/F10), LL/2 lung carcinoma (LL/2), CT26 colon carcinoma (CT26) and 4T1 breast cancer (4T1) cells in vitro. In vivo penfluridol was particularly effective at inhibiting LL/2 lung tumor growth, and obviously prolonged the survival time of mice bearing LL/2 lung tumors implanted subcutaneously. Accumulated unesterified cholesterol was found in all of the cancer cells treated with penfluridol, and this effect was most evident in LL/2, 4T1 and CT26 cells. No significant difference in serum cholesterol levels was found between the normal saline-treated mice and the penfluridol-treated mice. However, a dose-dependent decrease of total cholesterol in tumor tissues was observed in penfluridol-treated mice, which was most evident in B16/F10-, LL/2-, and 4T1-tumor-bearing mice. Conclusion: Our results suggested that penfluridol is not only cytotoxic to cancer cells in vitro but can also inhibit tumor growth in vivo. Dysregulation of cholesterol homeostasis by penfluridol may be involved in its anti-tumor mechanisms.

Interleukin-7 Enhances the in Vivo Anti-tumor Activity of Tumor-reactive CD8+ T cells with Induction of IFN-gamma in a Murine Breast Cancer Model

  • Yuan, Chun-Hui;Yang, Xue-Qin;Zhu, Cheng-Liang;Liu, Shao-Ping;Wang, Bi-Cheng;Wang, Fu-Bing
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.265-271
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    • 2014
  • Interleukin-7 (IL-7) is a potent anti-apoptotic cytokine that enhances immune effector cell functions and is essential for lymphocyte survival. While it known to induce differentiation and proliferation in some haematological malignancies, including certain types of leukaemias and lymphomas, little is known about its role in solid tumours, including breast cancer. In the current study, we investigated whether IL-7 could enhance the in vivo antitumor activity of tumor-reactive $CD8^+$ T cells with induction of IFN-${\gamma}$ in a murine breast cancer model. Human IL-7 cDNA was constructed into the eukaryotic expression plasmid pcDNA3.1, and then the recombinational pcDNA3.1-IL-7 was intratumorally injected in the TM40D BALB/C mouse graft model. Serum and intracellular IFN-${\gamma}$ levels were measured by ELISA and flow cytometry, respectively. $CD8^+$ T cell-mediated cytotoxicity was analyzed using the MTT method. Our results showed that IL-7 administration significantly inhibited tumor growth from day 15 after direct intratumoral injection of pcDNA3.1-IL-7. The anti-tumor effect correlated with a marked increase in the level of IFN-${\gamma}$ and breast cancer cells-specific CTL cytotoxicity. In vitro cytotoxicity assays showed that IL-7-treatment could augment cytolytic activity of $CD8^+$ T cells from tumor bearing mice, while anti-IFN-${\gamma}$ blocked the function of $CD8^+$ T cells, suggesting that IFN-${\gamma}$ mediated the cytolytic activity of $CD8^+$ T cells. Furthermore, in vivo neutralization of $CD8^+$ T lymphocytes by CD8 antibodies reversed the antitumor benefit of IL-7. Thus, we demonstrated that IL-7 exerts anti-tumor activity mainly through activating $CD8^+$ T cells and stimulating them to secrete IFN-${\gamma}$ in a murine breast tumor model. Based on these results, our study points to a potential novel way to treat breast cancer and may have important implications for clinical immunotherapy.

Enhancement of Anti-tumor Activity of Newcastle Disease Virus by the Synergistic Effect of Cytosine Deaminase

  • Lv, Zheng;Zhang, Tian-Yuan;Yin, Jie-Chao;Wang, Hui;Sun, Tian;Chen, Li-Qun;Bai, Fu-Liang;Wu, Wei;Ren, Gui-Ping;Li, De-Shan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7489-7496
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    • 2013
  • This study was conducted to investigate enhancement of anti-tumor effects of the lentogenic Newcastle disease virus Clone30 strain (NDV rClone30) expressing cytosine deaminase (CD) gene against tumor cells and in murine groin tumor-bearing models. Cytotoxic effects of the rClone30-CD/5-FC on the HepG2 cell line were examined by an MTT method. Anti-tumor activity of rClone30-CD/5-FC was examined in H22 tumor-bearing mice. Compared to the rClone30-CD virus treatment alone, NDV rClone30-CD/5-FC at 0.1 and 1 MOIs exerted significant cytotoxic effects (P<0.05) on HepG2 cells. For treatment of H22 tumor-bearing mice, recombinant NDV was injected together with 5-FC given by either intra-tumor injection or tail vein injection. When 5-FC was administered by intra-tumor injection, survival for the rClone30-CD/5-FC-treated mice was 4/6 for 80 days period vs 1/6, 0/6 and 0/6 for the mice treated with rClone30-CD, 5-FC and saline alone, respectively. When 5-FC was given by tail vein injection, survival for the rClone30-CD/5-FC-treated mice was 3/6 vs 2/6, 0/6 and 0/6 for the mice treated with rClone30-CD, 5-FC or saline alone, respectively. In this study, NDV was used for the first time to deliver the suicide gene for cancer therapy. Incorporation of the CD gene in the lentogenic NDV genome together with 5-FC significantly enhances cell death of HepG2 tumor cells in vitro, decreases tumor volume and increases survival of H22 tumor-bearing mice in vivo.

Antimicrobial Effect of Ethanol Extract of Dryopteris crassirhizoma Nakai on Propionibacterium acnes (관중(Dryopteris crassirhizoma Nakai) 추출물의 Propionibacterium acnes에 대한 항균 효과)

  • Yoon, Chang-Soon;Kim, Hyun-Ju;Lim, Hye-Won;Choi, Shin-Wook
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.32 no.3 s.58
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    • pp.201-208
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    • 2006
  • Propionibacterium acnes have been recognized as pus-forming bacteria triggering an inflammation in acne. The present study was conducted to evaluate antimicrobial activities of Dryopteris crassirhizoma Nakai against these etiologic agents of acne vulgaris and application possibility as a cosmetic resource. D. crassirhizoma crude extract and hexane fraction was prepared and its anti-acne effect against Propionibacterium acnes was investigated with minimum inhibitory concentration (MIC) and paper disk diffusion method. The MIC of D. crassirhizoma crude extract and hexane fraction was 0.008 mg/mL and 0.001 mg/mL, respectively. This implies that D. crassirhizoma extract may be an efficient anti-acne ingredient for cosmetics, as a crude extract. The paper disk diffusion assay showed that its anti-acne effect was similar to that of triclosan. The cytotoxic effect of D. crassirhizoma extract was determined by a colorimetric MTT assay using HaCaT cell line and D. crassirhizoma extract exhibited lower cytotoxic effects. Finally, we examine the stability of D. crassirhizoma extract to temperature and pH. The D. crassirhizoma extract was very stable to high temperatures ($25{\sim}121^{\circ}C$) and to wide pH range ($pH\; 2{\sim}11$), suggesting its utilization for cosmetics.

Studies on Constituents of Higher Fungi of Korea (LXXIII) Antitumor Components of the Cultured Mycelia of Paxillus atrotomentosus (한국산 고등 균류의 성분 연구(제73보) -좀우단버섯 배양 균사체의 항암 성분-)

  • Kwag, Sang-Deok;Bok, Jin-Woo;Hyun, Jin-Won;Choi, Eung-Chil;Kim, Byong-Kak
    • The Korean Journal of Mycology
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    • v.20 no.3
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    • pp.240-251
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    • 1992
  • To find antitumor components from higher fungi, the cultured mycelia of Paxillus atrotomentosus were extracted with hot water. The water soluble fraction was purified and separated by DEAE-cellulose ion exchange chromatography and Sepharose CL-4B gel filtration method. The separated fractions(Fr.) were designated CR A, B, C and D. Fr. A showed the highest inhibition ratio of 68.51% among the five tractions at a dose of 20 mg/kg/day. When Fr. A was examined for immunopotentiation activity, it increased the amount of the superoxide anion from activated macrophages to 1.1 fold and the number of plaques in hemolytic plaque assay to 2.3 fold, respectively. Otherwise, it did not show direct cytotoxity in sarcoma 180. Delayed type hypersensitiyity reaction showed that the decreased footpad swelling of tumor-hearing was restored to the normal. These results indicate that antitumor activity was exerted through immunopotentiation. Its chemical analysis showed 86.36% polysaccharide, 1.52% protein and 1.64% hexosamine. The polysaccharide consisted of fucose, galactose, glucose, mannose and xylose. This component was named paxillan.

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