• Food Science and Biotechnology
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• v.16 no.5
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• pp.858-862
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• 2007

The present study was conducted to examine the antioxidant activities and neuroprotective effects of methanolic extracts from Schizonepeta tenuifolia Briquet (STE). STE ($100\;{\mu}g/mL$) showed $43.33\;{\mu}M$ of total phenolic content, 64.43% of radical scavenging activity, and 0.157 of reducing power. In addition, the effect of STE on $H_2O_2$-induced DNA damage in human leucocytes was evaluated by the comet assay, where STE was a dose dependent inhibitor of DNA damage induced by $200{\mu}M$ of $H_2O_2$. The protective effect of STE against $H_2O_2$-induced oxidative damage on PC12 cells was investigated by an 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) reduction assay and lactate dehydrogenase (LDH) release assays. After 2 hr of cell exposure to $H_2O_2\;(500\;{\mu}M)$, a marked reduction in cell survival was observed. However, this reduction was significantly prevented by $1-50\;{\mu}g/mL$ of STE. Therefore, these results suggest that STE could be a new antioxidant candidate against neuronal diseases.

• Clinical and Experimental Reproductive Medicine
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• v.40 no.2
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• pp.67-75
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• 2013

Objective: To investigate the effect of peroxisome proliferator activated receptor ${\gamma}$(PPAR${\gamma}$) agonist on the cell proliferation properties and expression of human telomerase reverse transcriptase (hTERT) and aromatase in cultured endometrial stromal cell (ESC) from patients with endometriosis. Methods: Human endometrial tissues were obtained from women with endometriosis and healthy women (controls) using endometrial biopsy. Isolated ESCs were cultured and the cell proliferation was measured by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay and expression of hTERT, aromatase, and cyclooxygenase (COX)-2 by western blotting according to the addition of rosiglitazone (PPAR${\gamma}$ agonist). Results: We demonstrate that the cultured ESCs of endometriosis showed hTERT protein overexpression and increased cellular proliferation, which was inhibited by rosiglitazone, in a dose-dependent manner. At the same time, PPAR${\gamma}$ agonist also inhibited aromatase and COX-2 expression, resulting in decreased prostaglandin $E_2$ production in the ESCs of endometriosis. Conclusion: This study suggests that PPAR${\gamma}$ agonist plays an inhibitory role in the proliferative properties of eutopic endometrium with endometriosis by down-regulation of hTERT and COX-2 expression; this could be a new treatment target for endometriosis.

• YAKHAK HOEJI
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• v.43 no.6
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• pp.851-860
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• 1999

The effects of antimycin A(AA), dodium azide ($NaN_3$) and 2,4-dinitrophenol (DNP), which inhibit mitochondrial ATP production, on cellular functions of cultured astrocytes were studied. High concentrations of AA $(50{\;}\mu\textrm{g}/ml),{\;}NaN_3$ (100mM) and DNP (20mM) significantly decreased 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction, which was known to be related to mitochondrial function and then cel viability. AA ($50{\;}\mu\textrm{g}/ml$) increased lactate dehydrogenase (LDH) release and decreased [$^3H$] glutamate uptake, suggesting severe damage of cellular function by the concentrations of the compounds. Meanwhile, low concentrations of AA $(\leq{;\}10{\;}\mu\textrm{g}/ml),{\;}NaN_3{;\}(\leq{\;}50mM)$ and DNP ($\leq{\;}5mM$) significantly increased MTT reduction, the effect of which was specific to astrocytes. AA (5 and $10{\;}\mu\textrm{g}/ml$) did not affect LDH release and [$^3H$] glutamate uptake, indicating that these compounds increased MTT reduction at the low concentrations without cellular membrane damage. However, the low concentrations of AA produced significant decrease of MTT reduction in a glucose-free medium. Low concentrations of AA (1 and $5{\;}\mu\textrm{g}/ml$) did not change ATP production of astrocytes in the medium containing 10 mM glucose, but completely inhibited in a glucose-free medium, suggesting marked increase of cytosolic ATP production by the blockade of mitochondrial ATP production with low concentrations of AA. These results suggest that astrocytes have ability to enhance neuronal function or survival under conditions of incomplete ischemia or early by enhancement of glycolysis, and that cellular reduction of MTT occurs not only mitochondrially but also extramitchondrially.

• YAKHAK HOEJI
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• v.40 no.2
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• pp.218-224
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• 1996

This laboratory has recently reported the synthesis and in vitro antitumor activity of PT(II) complexes containing ethylenediamine and diphosphine. In view of the reports of others, cisplatin is toxic to the kidney since the kidney's vulnerability to PT(II) complexes may originate in its ability to accumulate and retain platinum to a greater degree than other organs. The in vitro cytotoxicity of these synthetic PT(II) complexes on the primary cultured proximal tubular cells of rabbit kidney and renal cortical cells of human kidney was investigated. Three endpoints for cytotoxicity tests were evaluated:3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), $^3H$-thymidine uptake and the glucose consumption tests. The rank order of sensitivity exhibited $^3H$-thymidine uptake>MTT>glucose consumption test. The agents with diphosphine leaving group were significantly less cytotoxic than cisplatin. Moreover, 1,2-bis(diphenylphosphino)ethane (DPPE) exhibited less cytotoxicity than 1.3-bis (diphenylphosphino)propane (DPPP) against on rabbit and human cultured kidney cells. Based on these results, the decreased nephrotoxicity of these new complexes over cisplatin appeared to be partially attributable to a leaving group of DPPP and DPPE. This novel class of platinum compound represents a valuable lead in the development of a "third-generation" agent.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.10
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• pp.5905-5910
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• 2013

Background: To investigate the preventive and therapeutic potential of garlic oil on human pancreatic carcinoma cells. Methods: The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay was performed to study the effects of garlic oil on three human pancreatic cancer cell lines, AsPC-1, Mia PaCa-2 and PANC-1. Cell cycle progression and apoptosis were detected by flow cytometry (FCM), staining with PI and annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI), respectively. Morphologic changes of pancreatic cancer cells were observed under transmission electron microscopy (TEM) after treatment with garlic oil at low inhibitory concentrations ($2.5{\mu}M$ and $10{\mu}M$) for 24 hours. Results: Proliferation of the AsPC-1, PANC-1, and Mia PaCa-2 cells was obviously inhibited in the first 24 hours with the MTT assay. The inhibition effect was more significant after 48 hours. When cells were exposed to garlic oil at higher concentrations, an early change of the apoptotic tendency was detected by FCM and TEM. Conclusion: Garlic oil could inhibit the proliferation of AsPC-1, PANC-1, and Mia PaCa-2 cells in this study. Moreover, due to programmed cell death, cell cycle arrest, or both, pro-apoptosis effects on AsPC-1 cells were induced by garlic oil in a dose and time dependent manner in vitro.

• Restorative Dentistry and Endodontics
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• v.45 no.4
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• pp.47.1-47.11
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• 2020

Objectives: The aim of this study was to assess the physicochemical properties, cytotoxicity and penetration into dentinal tubules of ChlorCid^TM Surf (3% sodium hypochlorite [NaOCl] with surfactant) in comparison to ChlorCid^TM (3% NaOCl without surfactant). Materials and Methods: The physicochemical properties evaluated were pH, surface tension, free available chlorine (FAC) and contact angle. Cytotoxicity was evaluated in L929 fibroblasts exposed to the solutions by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and neutral red assays. Assessment of penetration into dentinal tubules was performed by staining single-rooted permanent human teeth with crystal violet (n = 9), which were irrigated with the solutions and analyzed in cervical, middle and apical segments. Data were analyzed by one-way analysis of variance (ANOVA) and Tukey's post-test, 2-way ANOVA and Bonferroni's post-test or t-test (α = 0.05). Results: ChlorCid^TM Surf and ChlorCid^TM FAC values were close to those indicated by the manufacturer. ChlorCid^TM Surf showed lower surface tension and contact angle on dentin, and higher pH than ChlorCid^TM (p < 0.05). The penetration of ChlorCid^TM Surf was higher in cervical and middle segments, compared with ChlorCid^TM (p < 0.05). There was no difference in irrigant cytotoxicity (p > 0.05). Conclusions: ChlorCid^TM Surf showed lower surface tension, lower contact angle on root canal dentin, higher penetration into dentinal tubules and more alkaline pH, compared with ChlorCid^TM. However, both solutions showed similar cytotoxicity and FAC content.

• Advances in Traditional Medicine
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• v.1 no.1
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• pp.45-54
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• 2000

Cannabidiol derivatives (1, 2 and 3), and 5-fluorouracil (4, 5-FU) were tested for their growth inhibitory effects against human oral epitheloid carcinoma cell lines (KB) using two different 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and sulforhodamine B protein (SRB) assay. These compounds showed a potent inhibitory activity in vitro in the micromolar range against KB cell lines. In general, the antitumor activity of these compounds (1, 2, 3 and 4) was in a dose-dependent over the micromolar concentration ranges from $1\;{\mu}M\;to\;100\;{\mu}M$. The comparison of $IC_{50}$ values of these compounds in tumor cell lines shows that their susceptibility to these compounds decreases in the following order: CBD > 5-FU > CBDME > CBDDE by the MTT assay and SRB assay. Cannabidiol derivatives (1, 2 and 3), and 5-FU were tested for their cytotoxic effects on NIH 3T3 fibroblasts using two different MTT assay and SRB assay. These compounds exhibited potent cytotoxic activities in vitro in the micromolar range against NIH 3T3 fibroblasts. In general, the cytotoxic activities of these compounds (1, 2, 3 and 4) were in a dose-dependent over the micromolar concentration range $1\;{\mu}M\;to\;100\;{\mu}M$. The comparison of $CD_{50}$ values of these compounds on NIH 3T3 fibroblasts shows that their susceptibility to these compounds decreases in the following order; CBD > 5-FU > CBDDE > CBDME by MTT assay, CBD > 5-FU > CBDME > CBDDE by SRB assay. These results suggest that cannabidiol (1, CBD) retains the most growth-inhibitory activity against KB cell lines.

• Korean Journal of Pharmacognosy
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• v.28 no.4
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• pp.264-270
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• 1997

This Study was carried out develop antitumor effect of the n-butanol soluble of fraction of Perilla frutescens on human skin melanoma cells. The antitumor activity of various fractions obtained form n-butanol soluble fraction of Perilla frutescens was evaluated in human skin melanoma cells. The antitumor activity of the n-butanol soluble fraction in human skin melanoma cells was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, neutral red (NR) assay and sulforhordamine B protein (SRB) assay of colorimetic assay methods. The light microscopic study was carried out to observe morphological changes of cultured human skin melanoma cells. These results were obtained follows; The fractions 5 and 6 of the n-butanol soluble fraction of P frutescens were shown significant antitumor activities. The number of human skin melanoma cells were decreased and tend to form cell cluster by treatment with actions 5 and 7 of the n-butanol soluble fraction of P. frutescens. The fraction 6 of the the n-butanol soluble fraction showed the highest antitumor activity on P. frutescens. These results suggest that the fraction 6 of the n-butanol soluble fraction of P. frutescens may be a valuable choice for the studies on the treatment of human skin tumors.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.453-456
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• 2001

The purpose of this research was to investigate the effects of extracts from cultured hairy roots of R. undulatum on human kidney epithelial cells. Hairy roots were induced by a co-culture with A. rhizogenes ATCCl5834 and cultured in WPM medium. The cytotoxicity was measured by colorimetric assay using 3-(4,5-dimethythiazol-2-yl)-2,5- diphenyl-2H -tetrazolium bromide (MTT), neutral red (NR) and sulforhodamine protein B (SRB) with human kidney epithelial cell lines A498. MTT, NR and SRB quantities decreased propotionally in cultured A498 cells treated with the water or chloroform extracts of cultured hairy roots at increasing concentrations. These results suggest that extracts of cultured hairy 개ots are cytotoxic on human epithelial cells. The cytotoxicity of chloroforrm fraction was stronger than that of water fraction. The values of $MTT_{50}$, $NR_{50}$, $SRB_{50}$ of the extracts of chloroform fraction and those of water fraction were measured to be 289.3${\mu}g$/ml, 302.7${\mu}g$/ml. 433.8${\mu}g$/ml and 475.8${\mu}g$/ml. 428.3${\mu}g$/ml, 549.5${\mu}g$/ml in A498 cell line.

• Journal of Ginseng Research
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• v.44 no.2
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• pp.215-221
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• 2020

Background: Panax ginseng has been used for a variety of medical purposes in eastern countries for more than two thousand years. From the extensive experiences accumulated in its long medication use history and the substantial strong evidence in modern research studies, we know that ginseng has various pharmacological activities, such as antitumor, antidiabetic, antioxidant, and cardiovascular system-protective effects. The active chemical constituents of ginseng, ginsenosides, are rich in structural diversity and exhibit a wide range of biological activities. Methods: Ginsenoside constituents from P. ginseng flower buds were isolated and purified by various chromatographic methods, and their structures were identified by spectroscopic analysis and comparison with the reported data. The 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H- tetrazolium bromide method was used to test their cytotoxic effects on three human cancer cell lines. Results: Six ginsenosides, namely 6'-malonyl formyl ginsenoside F₁ (1), 3β-acetoxyl ginsenoside F₁ (2), ginsenoside Rh₂₄ (6), ginsenoside Rh₂₅ (7), 7β-hydroxyl ginsenoside Rd (8) and ginsenoside Rh₂₆ (10) were isolated and elucidated as new compounds, together with four known compounds (3-5 and 9). In addition, the cytotoxicity of these isolated compounds was shown as half inhibitory concentration values, a tentative structure-activity relationship was also discussed based on the results of our bioassay. Conclusion: The study of chemical constituents was useful for the quality control of P. ginseng flower buds. The study on antitumor activities showed that new Compound 1 exhibited moderate cytotoxic activities against HL-60, MGC80-3 and Hep-G2 with half inhibitory concentration values of 16.74, 29.51 and 20.48 μM, respectively.