• Title/Summary/Keyword: MTT test

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Effect of Podophyllotoxin Conjugated Stearic Acid Grafted Chitosan Oligosaccharide Micelle on Human Glioma Cells

  • Wang, Geng Huan;Shen, He Ping;Huang, Xuan;Jiang, Xiao Hong;Jin, Cheng Sheng;Chu, Zheng Min
    • Journal of Korean Neurosurgical Society
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    • v.63 no.6
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    • pp.698-706
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    • 2020
  • Objective : To study the physiochemical characteristics of podophyllotoxin (PPT) conjugated stearic acid grafted chitosan oligosaccharide micelle (PPT-CSO-SA), and evaluate the ability of the potential antineoplastic effects against glioma cells. Methods : PPT-CSO-SA was prepared by a dialysis method. The quality of PPT-CSO-SA including micellar size, zeta potential, drug encapsulation efficiency and drug release profiles was evaluated. Glioma cells were cultured and treated with PPT and PPT-CSO-SA. The ability of glioma cells to uptake PPT-CSO-SA was observed. The proliferation of glioma cells was determined by 3-[4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay. The apoptosis and morphology of U251 cells were observed by 4',6-Diamidino-2-phenylindole dihydrochloride (DAPI) dye staining. Cell cycle analysis was performed by flow cytometry. The migration ability of U251 cells was determined by wound healing test. Results : PPT-CSO-SA had nano-level particle size and sustained release property. The encapsulation efficiency of drug reached a high level. The cellular uptake percentage of PPT in glioma cells was lower than that of PPT-CSO-SA (p<0.05). The inhibitory effect of PPT-CSO-SA on glioma cells proliferation was significantly stronger than that of PPT (p<0.05). The morphologic change of apoptosis cell such as shrinkage, karyorrhexis and karyopyknosis were observed. The percentage of U251 cells in G2/M phase increased significantly in the PPT-CSO-SA group compared with PPT group (p<0.05). Compared with the PPT group, the cell migration ability of the PPT-CSO-SA group was significantly inhibited after 12 and 24 hours (p<0.05). Conclusion : PPT-CSO-SA can effectively enhance the glioma cellular uptake of drugs, inhibit glioma cells proliferation and migration, induce G2/M phase arrest of them, and promote their apoptosis. It may be a promising anti-glioma nano-drug.

Inhibition of Cell Proliferation and Induction of Apoptosis by Ethanolic Extract of Lespedeza cuneata G. Don in Human Colorectal Cancer HT-29 cells (야관문의 에탄올 추출물에 의한 대장암세포의 성장억제 및 세포사멸유도)

  • Zhao, Qian;Kim, Yeah-Un;Han, In-Hwa;Yun, Jung-Mi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.6
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    • pp.911-917
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    • 2016
  • Lespedeza cuneata G. Don is an edible perennial herb used in traditional Korean medicine. We investigated the anti-proliferative properties and mechanism of L. cuneata extract. The ethanolic extract of L. cuneata dose-and time-dependently inhibited human colorectal cancer cell proliferation. A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay was used to test the effect of the extract on proliferation of HT-29 colorectal cancer cells. The extract inhibited HT-29 cell proliferation with an $IC_{50}$ value of $554.26{\pm}8.81{\mu}g/mL$. L. cuneata extract suppressed production of pro-inflammatory cytokines interleukin-6 and tumor necrosis $factor-{\alpha}$. Apoptosis was evaluated by analysis of DNA fragmentation, poly(ADP-ribose) polymerase cleavage, caspase-3 activity, and protein expression of pro-apoptotic (Bax) and anti-apoptotic (Bcl-2). Our results demonstrated that the extract induced DNA fragmentation and characteristic morphological changes associated with apoptosis in HT-29 colorectal cancer cells. The extract also time- and dose-dependently up-regulated expression of the Bax and down-regulated expression of the Bcl-2. Furthermore, the extract dose- and time-dependently enhanced caspase-3 activity. Our findings provide evidence that L. cuneata extract may mediate its anti-proliferative effect via modulation of apoptosis.

Antibacterial, antioxidant and antitumor activities of mushroom mycelium mixed culture extracts (버섯 균사체 혼합 배양 추출물의 항균, 항산화 및 항암 활성)

  • Kim, Man-Cheol;Kim, Ju-Sang;Heo, Moon-Soo
    • KSBB Journal
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    • v.23 no.2
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    • pp.158-163
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    • 2008
  • In this study, we have investigated the antibacterial, antioxidant, and antitumor activities of mycelium cultural extract from mushroom. Mushroom mycelium was grown in a synthetic liquid media such as PD broth, YM broth or citrus extracts. In antibacterial activity test, the best result was achieved when mycelium cultural extracts from Phellinus linteus and Coriolus versicolor were incubated together on YM broth. On the other hand, mushroom mycelium cultured on citrus extracts showed better activity than that on PD broth. We have also tested the antioxidant activity at concentration up to 10 mg of mycelium cultural extract/mL. The more it is in higher concentration, the more the activity increases. The higher antioxidant activity was observed both on PD broth containing the Phellinus linteus and Coriolus versicolor mycelium and citrus extract containing the same. The complex culture extracts obtained from the synthetic medium and citrus extract medium showed 10-89% of the 1,1-diphenyl-2-picrylhydrazyl radical scavenger activity. The antitumor activity of mycelium cultural extract was examined by using MTT assay on A549 cells. Mushroom mycelium cultured on citrus extracts showed interestingly higher antitumor activity than that on synthetic liquid media.

Inhibitory Effect of YBR on Hepatic Fibrogenesis (YBR의 간섬유화(肝纖維化)억제 효과(效果)에 관한 연구(硏究))

  • Seung, Hyun-Seok;Woo, Hong-Jung
    • The Journal of Internal Korean Medicine
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    • v.31 no.2
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    • pp.314-330
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    • 2010
  • Objective : This study was performed to investigate the anti-fibrogenic effect and changes of inflammation-related genes by YBR I and YBR II (YBR I: Arteisiae Capillaris Herba, Atractylodis Rhizoma Alba, Hoelen/ YBR II: YBR I +Sanguisorbae Radix, Biotae Cacumen, Cirsii Japonici Herba) on HSC(hepatic stellate cells)-T6 and TAA-induced rat liver tissue. Materials and Methods : HSC-T6 were treated with various concentrations of distilled-water extract YBR I and YBR II extract for 24, 48 and 72 hours. After the treatment, cell viability, proliferation, procollagen levels and IL-6 levels were measured by using MTT Assay, BrdU Assay, Procollagen Type 1 C-peptide EIA kit, and Murine IL-6 ELISA Development kit. Rat liver fibrosis was induced by intraperitoneal TAA injection of 150mg/kg 3 times a week for 6 weeks. After the treatment, body weight, liver & spleen weights, liver function test, complete blood cell count and change of portal pressure were studied. In addition, gene expressions of ASMA, IL-6, MMP-2, TIMP-1 and TIMP-2, all of which are known to be associated with liver fibrosis, were analyzed by using Real-Time PCR. After YBR I and YBR IItreatment, percentages of collagen in TAA-induced rat liver tissue were measured. Results : The viability and proliferation of the HSC-T6 decreased as the concentration increased. The production of procollagen decreased as the concentration increased. The production of IL-6 was little influenced by YBR I and YBR II. There was no difference in rat body weight between the TAA-only group and the YBR groups. Compared with rat liver weight of TAA-only group, that of the YBR groups increased. In the YBR I group, the serum level of AST elevated by TAA injection significantly decreased and in the YBR I and II group, the serum level of ALP and ALT elevated by TAA injection decreased. In the YBR I group, white blood cell count elevated by TAA injection decreased but platelets increased. In the YBR I group, the portal pressure elevated by TAA injection significantly decreased. Decreases in the gene expression of ASMA and MMP-2 were observed in the YBR I group. The gene expression of IL-6 was little influenced by YBR I and YBR II -treated groups. In the histological finding, TAA injections caused severe fibrosis, but YBR I and YBR II treatment significantly reduced the amounts of hepatic collagens. Conclusions : These results suggest that YBR I and II have inhibitory effects on the hepatic fibrogenesis.

Protective Effects of Bojungmyunyuk-dan in Cisplatin Treated Brain Cell Death (Cisplatin에 의한 뇌세포사멸에서 보중면역단의 방어효과)

  • Yoo Kyung Tae;Moon Seok Jae;Won Jin Hee;Kim Dong Woung;Lee Jong Deok;Won Kyoung Sook;Moon Goo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.394-402
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    • 2003
  • This study was designed to investigate the protective effect of Bojungmyunyuk-dan(BJMY-Dan) on the cisplatin-induced cytotoxicity of primary rat astrocytes. BJMY-Dan is an oriental herbal prescription for its ability to recover protective effects against anti-cancer chemotherapies. After astrocytes were treated cisplatin, MTT assay was performed for cell viability test. To explore the mechanism of cytotoxicity, I used the several measures of apoptosis to determine whether this processes was involved in cisplatin-induced cell damage in astrocytes. Also, astrocytes were treated with BJMY-Dan and then, followed by the addition of cisplatin. Cisplatin decreased the viability of astrocytes in a dose and time-dependent manner. BJMY-Dan increased the viability of astrocytes treated cisplatin. Astrocytes treated cisplatin were revealed as apoptosis characterized by nuclear staining and flow cytometry. BJMY-Dan protected astrocytes from cisplatin-induced nuclear fragmentation and chromatin condensation. Also, caspase-3 and caspase-9 proteases were activated in astrocytes by cisplatin. BJMY-Dan inhibited the activation of caspase proteases in cisplatin-treated astrocytes. Cleavage of [poly(ADP-ribose) polymerase](PARP) was occurred at 12hr after treatment of cisplatin in astrocytes. BJMY-Dan recovered the cleavage of PARP in cisplatin-treated astrocytes. Also, BJMY-Dan inhibited the activation of pro-apoptotic factor, Bak by cisplatin. Lastly, astrocytes stained with JC-1 and Rhodamine 123 were photographed by fluorescence microscope to visualize changes of mitochondrial membrane permeability transition(MPT) during treatment with cisplatin for 24hr. BJMY-Dan recovered the change of MPT by cisplatin in astrocytes. According to above results, BJMY-Dan may protect astrocytes from cytotoxicity induced by chemotherapeutic agents, including cisplatin.

Differential Cytotoxic Effects of Jaspine B in Various Cancer Cells (다양한 암세포주에서 Jaspine B의 함암활성 비교)

  • Lee, Jihoon;Choi, Kwangik;Kwon, Mihwa;Lee, Dongjoo;Choi, Min-Koo;Song, Im-Sook
    • Journal of Life Science
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    • v.26 no.12
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    • pp.1392-1399
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    • 2016
  • Jaspine B is an anhydrophytosphingosine that is isolated from a marine sponge. Because of its structural similarity to sphingosine, it shows anti-cancer effects in human carcinomas. Therefore, this study aims to investigate its anti-proliferative effect on various cancer cells and to correlate its association with the intracellular accumulation of Jaspine B in relevant cancer cells. The anti-proliferative effect of Jaspine B in various cancer cells was determined by a cell viability test, and the intracellular concentration of Jaspine B in relevant cancer cells was determined using mass spectrometry coupled with liquid chromatography. The correlation coefficient and p value between the cytotoxicity and the cell accumulation of Jaspine B were determined using SPSS 16.1. The cytotoxicity of Jaspine B varied depending on the type of cancer cell when compared the $EC_{50}$ values of Jaspine B. Breast and melanoma cancer cells were susceptible to Jaspine B, whereas renal carcinoma cells were resistant. The intracellular concentrations of Jaspine B had a reciprocal correlation with the $EC_{50}$ values in the same cells (r = 0.838). The results suggested that the anti-proliferative effect of Jaspine B was associated with the cellular accumulation of this compound. However, Jaspine B was not a substrate for P-glycoprotein and breast cancer resistance protein, as major efflux pumps caused multidrug resistance. The maintenance of a high intracellular concentration is crucial for the cytotoxic effect of Jaspine B; however, efflux pumps may not be a controlling factor for Jaspine B-related resistance in cancer cells.

Protective Effects of Bojungbangam-tang Extracts on ECV304 Cell Cytotoxicity (보정방암탕 추출물의 혈관내피세포독성에 대한 방어효과)

  • Kwon, Kang-Beom;Kim, Eun-Kyung;Song, Mi-Young;Han, Mi-Jeong;Lee, Su-Yeop;Lee, Heon-Jae;Lee, Young-Rae;Ju, Sung-Min;Ryu, Do-Gon;Kim, Sung-Hoon;Jeon, Byung-Hun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.2
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    • pp.404-407
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    • 2007
  • This study was designed to investigate the protective effect of Bojungbangam-tang Ethanol Extracts (EBJT) on cisplatin and hydrogen peroxide-induced cytotoxicity of human endothelial cell line ECV304 cells. After cells were treated with cisplatin and hydrogen peroxide, MTT assay was performed for cell viability test. To explore the mechanism of cytotoxicity, we used the several measures of apoptosis to determine whether this processes was involved in cisplatin and hydrogen peroxide-induced cell damage in ECV304 cells. Also, cells were treated with EBJT and then, followed by the addition of cisplatin or hydrogen peroxide. Cisplatin or hydrogen peroxide decreased the viability of ECV304 cells in a dose-dependent manner. ECV304 cells treated cisplatin or hydrogen peroxide were revealed as apoptosis characterized by nuclear staining. EBJT protected ECV304 cells from cisplatin or hydrogen peroxide-induced nuclear fragmentation and chromatin condensation. Also, EBJT inhibited the cleavage of poly(ADP-ribose) polymerase (PARP) in cisplatin or hydrogen peroxide-treated ECV304 cells. According to above results, EBJT may protect ECV304 cells from the apotosis induced by cisplatin or hydrogen peroxide.

Effect of DanGuiBoHyulTangGami-Bang on the Alopecia and Hair Growth Stimulation (當歸補血湯加味方이 모발탈락 및 성장에 미치는 영향)

  • Kim, Phill-soo;Kim, Hee-taek;Roh, Seok-sun;Hwang, Chung-yeon
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.17 no.3
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    • pp.38-60
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    • 2004
  • We made an experiment if the extracts of DanGuiBoHyulTangGami-Bang(DBTG) and 15 kinds of the medical herbs used the materials of DBTG were effective on the hair formation palpation and the falling out of hair, and came to the following conclusions. 1. The extracts of Paeonia lactiflora, Cuscuta chinensis and Angelica tenuissima of DBTG consisted of the 15 kinds of the medical herbs kept the activity of 5${\alpha}$-reductase type Ⅱ from being active 75.3$\%$, 63.8$\%$. 75.5$\%$. 2. The hair growth index, 1.6(control group 0.8) of the extracts of DBTG bas a little effect on the hair growth palpation and that of Rubus coreanus 1.8(control group 0.4) was the most effective one of the medical herbs, and Paeonia lactiflora 2.3(control group 1.7) and Vitex rotundifolia 2.3(control group 1.5) showed the effect on hair formation palpation. 3. The hair growth period couldn't be extended by DBTG in this experimental stage. 4. The 15 kinds of constitution medicines of DBTG didn't have effects in dermal papilla cells DNA increase, IGF- I, KGF, HGF the revelation of a gene heredity, the protein synthesis of the hair follicle tissues. 5. All of the 15 kinds of constitution medicines of DBTG didn't have the antibacterial activity in Paper disc rule. 6. The results from the test of a radical scavenging activity of the 15 kinds of constitution medicines of DBTG showed that the extracts of Paeoria lactiflora, Scutellaria baicalensis, Rubus coreanus have the superior antioxidant activity in the concentration of 0.01$\%$ and 0.001$\%$ 7. In the formation controlled experiment, Vitex rotundifolia (70.6$\%$), Scutellaria baicalensis (47.1$\%$, Saposhnikovia (44.8$\%$) of the 15 kinds of constitution medicines of DBTG in the 50㎍/㎖ concentration controled NO forming and Vitex rotundifolia (12.7$\%$) controled NO forming in the 5㎍/㎖ concentration in order. 8. MTT(lC/50) of the extracts of Rehmannia glutinosa, Paeonia lactiflora, Scutellaria baicalensis, Lycium chinense, Rubus coreanus of the 15 kinds of constitution medicines of DBIG was more than 500㎍/㎖ and had the least cell virulence.

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Physiological Activities using Root and Stem Extracts of Cymbidium (심비디움 뿌리 및 줄기 추출물의 생리 활성)

  • Kim, Hye-Ran;Park, Gyu-Nam;Jung, Bo-Kyoung;Shin, Yu-Su;Chang, Kyung-Soo
    • Journal of the Korean Applied Science and Technology
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    • v.33 no.4
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    • pp.848-854
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    • 2016
  • Cymbidium is one of perennial herbs belonging to the Orchidaceae and is known as a medicinal plant. However, its scientific data are insufficient. The purpose of this study is to extract from root and stem of Cymbidium, to investigate the biological effects of them. Cymbidium antibacterial effects of the extracts were performed by antibacterial test against Staphylococcus aureus (S. aureus), Staphylococcus saphrophyticus (S. saprophyticus), Proteus vulgaris (P. vulgaris) and Klebsiella pneumonia (K. pneumonia). Antioxidant effects of the extracts were carried out by DPPH radical scavenging. Total phenolic contents were also determined. Moreover, Cell viability of extracts against MTT assay was cell viability against HepG2 cell and also measures Cholesterol adsorptivity of extracts. In this study, the extracts inhibited the growth of bacteria. Particularly Cymbidium root extracts by only ethanol extraction showed highest antimicrobial effect against S. aureus. The Cymbidium stem extracts by both ethanol extraction and sonication for 1 hour had higher antioxidant activites as well as total phenolic contents. Cell cytotoxicity showed higher than $50{\mu}g/mL$. Cholesterol adsorptivity showed lower than 20%. These results suggest that the Cymbidium might be a source of anti-bacterials and anti-oxidants.

The effects of Naetakchunkeum-san on the Allergic Contact Dermatitis induced by DNCB (내탁천금산(內托千金散)이 DNCB로 유발된 알레르기성 접촉피부염에 미치는 영향)

  • Kim, Joon-Jeong;Kim, Hee-Taek
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.22 no.1
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    • pp.46-61
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    • 2009
  • Background and Objectives : Allergic Contact Dermatitis is the disease affected by industrialization. The more industrialization advanced, the more materials that could induce the allergic contact dermatitis have been increased. Therefore in oriental medicine, various studies have been performed. The objective of this study is to investigate the effects of Naetakchunkeum-san on the Allergic Contact Dermatitis induced by 2,4-dinitro-chlorobezene(DNCB). Meterial and Methods : Twenty eight mice were divided into four groups ; normal, control, experimental group A and B. Control and experimental group were induced allergic contact dermatitis by DNCB. Experimental group A was orally administered the Naetakchunkeum-san and experimental group B was orally administered the prednisolone. In this study, ear thickness measurement, observation auricle microphotograph, Myeloperoxidase(MPO) activity measurement, Reverse transcription-polymerase chain reaction(RT-PCR) analysis of the mRNA level of $TNF-\alpha$, $IL-1\beta$, $INF-\gamma$ were performed on these four groups. In addition, the effect of Naetakchunkeum-san on cell viability and the effect of Naetakchunkeum-san on the compound 48/80-induced histamine release from HMC and RPMC were measured, Results : 1. In contact hypersensitivity assay, experimental group A and B showed decreased ear thickness compared with control group, 2. In experimental group A, pathological lesion of dermatitis were alleviated. In addition, the numbers of infiltrated cells were reduced, and cleft was not shown compared with control group, In experimental group B, similar results were shown. 3. There was a significant increase in MPO activity in control group compared with normal group, Experimental group A and B significantly inhibited the increase in MPO activity compared with control group. 4, The level of expression of $TNF-\alpha$, $IL-1\beta$, $INF-\gamma$ in experimental group A and B were significantly lower than those in control group. As the internal control, cyclophilin mRNA was also reverse-transcribed and amplified. 5, In MTT assay, there were no statistically significant differences in 100 ${\mu}g/ml$, 200 ${\mu}g/ml$, 500 ${\mu}g/ml$, 1000 ${\mu}g/ml$ Naetakchunkeum-san treated group from 0 ${\mu}g/ml$ Naetakchunkeum-san treated group as determined by the Tukey test. 6. Naetakchunkeum-san dose-dependently inhibited the compound 48/80-induced histamine release from both HMC and RPMC. Conclusions : According to above experiments, Naetakchunkeum-san may be applied to allergic contact dermatitis.

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