• Title/Summary/Keyword: MTT quantity

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Studies on the Cytotoxicity and Antitumor Activity of Perilla frutescens (소엽의 세포독성 및 항암작용에 관한 연구)

  • Han, Du-Seok;Chung, Boung-Ho;Yoo, Hyeon-Gyeong;Kim, Young-Ok;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.25 no.3
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    • pp.249-257
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    • 1994
  • The cytotoxic and antitumor activity of Perilla frutescens extract on cultured 3T3 fibroblast and skin melanoma cells were evaluated by tetrazolium MTT (MTT) and neutral red (NR) colorimetric assay methods. Lactate dehydrogenase activity was also measured. The light microscopic study was carried out to observe morphological changes of cultured mouse fibroblast and skin melanoma cells. The results were as follows: 1. Water and ether extracts showed a significant cytotoxicity in 3T3 fibroblast and all extracts exhibited a significant anti-tumor activity in skin melanoma cells. Methanol, ethyl acetate and ethanol extracts showed low cytotoxic effects, but exhibited a high anti-tumor activity. 2. The MTT absorbance in 3T3 fibroblast was significantly decreased by treatment with ether, water, chloroform and ethanol extracts and skin melanoma cells was significantly decreased by treatment with all extracts. The difference in MTT absorbance in two cell types was most remarkable when treated with methanol and ethanol extracts. 3. Methanol and ethyl acetate extracts showed the strongest effect in growth inhibition of melanoma cells. These results indicated that methanol extract possessed a low cytotoxicity and a strong anti-tumor activity.

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Galactosylated PEI-PEG as nonviral gene transfer agent for hepatocyte targeting and imaging probe

  • Kim, Eun-Mi;Oh, In-Joon;Jeong, Hwan-Jeong;Shin, Sang-Chul;Lee, Yong-Bok
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.248.3-249
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    • 2003
  • Objectives: Galatosylated PEI was synthesized and characterized for gene delivery to hepatocytes. It was modified by conjugating with hydrophilic PEG to improve in vivo circulation. And we studied the possibility as an imaging modality for monitoring of gene delivery using gal-PEI derivatives. Methods: The substitution values of galactose in PEI were calculated by resorcinol/sulfuric acid method and quantity of PEG was calculated by comparing NMR peak. Cytotoxicity was determined by MTT. (omitted)

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The Effects of BunSimGiEumGami-Bang(Fenxinqiyinjiameifang) on Serotonin of P815 cell (분심기음가미방(分心氣飮加味方)의 항산화능과 serotonin 대사에 미치는 영향)

  • Lim, Jae-Won;Jung, In-Chul;Lee, Sang-Ryong
    • Journal of Oriental Neuropsychiatry
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    • v.22 no.2
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    • pp.147-162
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    • 2011
  • Objectives : This experiment was designed to investigate the effect of the BSGE hot water extract on serotonin biosynthesis of depression model. Methods : The cytotoxicity of the BSGE hot water extract was analyzed by MTT assay on P815 cell. The antioxidant activity was measured by DPPH free radical-scavenging activity and SOD activity on P815 cell. The quantity of 5-HT and expression of TPH-1, AAADC and MAOa mRNA was measured by of HPLC profle Analysis on P815 cell. Results : 1. The BSGE hot water extract increased DPPH free radical-scavenging activity and SOD activity on P815 cell. 2. The BSGE hot water extract increased significantly the quantity of 5-HT. 3. The BSGE hot water extract increased the expression of TPH-1 mRNA. Conclusions : This experiment shows that the BSGE hot water extract might be effective for the prevention and treatment of depression. Investigation into the clinical use of the BSGE hot water extract for depression is suggested for future research.

The Protective Effects of EGCG Extracted from Green Tea on Apoptosis Induced by$H_2O_2$ in Conjunctival Cell Lines ($H_2O_2$에 의한 결막 세포주의 세포고사에서 녹차추출물 EGCG의 보호효과)

  • Park, Su-Kyoung;Chae, Soo-Chul;Kho, Eun-Gyeong;Ryu, Geun-Chang;Kim, Jai-Min;Na, Myung-Suk;Lee, Jong-Bin
    • Journal of Korean Ophthalmic Optics Society
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    • v.13 no.3
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    • pp.95-101
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    • 2008
  • Purpose: Hydrogen peroxide which is one of the reactive oxygen species has been seen to cause various diseases, various cellular disinfections, gene transformation and cell death. The goals of this study were to determine the protective effect of EGCG against $H_2O_2$-induced apoptotic death in conjunctival cell lines. Methods: We measured cell viability by MTT assay and analyzed DNA fragmentation to check up a distinctive feature in cell death and measured the removal ability of free radicals by DPPH free radical scavenging assay and evaluated the oxygen free radical's quantity in the cell by DCFH-DA assay. The mRNA expression in the cell were examined by RT-PCR. Results: Cell viability and free radical scavening activites was significantly increased in dose dependently after cell was exposed to EGCG. And DNA fragmentation and intracellular ROS was decreased. It was showed the mRNA expression which increase of bcl-2, bcl-xL expression and decrease of bax expression. Conclusions: From these results, it suggests that EGCG has an antioxidant effect and protects conjunctival cell lines from the $H_2O_2$-mediated apoptosis through the modulation of the mRNA expression.

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Study on the promotion of inflammation and whitening of natural materials using bioconversion technology

  • Lee, Se-Won;Lim, Jeong-Muk;Lee, Seong-Hyeon;Lee, Jeong-Ho;Oh, Byung-Teak
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.116-116
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    • 2018
  • Bioconversion, the enzymatic process by microorganism on organic precursor to desired products. The natural extract is converted into a form that can be easily absorbed into the skin, while scaling up of to higher quantity is possible. Selection of naturally processed raw material rather than chemically processed is preferred. Therefore, fermentation was carried out by mixing Rubus coreanus Miquel, soybean, Zanthoxylum schinifolium as bioconverting materials, the possibility of inflammation, whitening material were checked. In this study, useful microorganisms were isolated from various salted fish, and 16S rDNA sequence was analyzed to confirm their genetic characteristics. Combining the three natural materials using bioconversion technology to study their activity before and after fermentation. To evaluate the antioxidant activity and the active ingredient content the DPPH radical scavenging activity and the total polyphenol content were examined. Raw 264.7 cells were used to evaluate MTT assay, NO and $TNF-{\alpha}$ production inhibitory activity. Also, to evaluate the whitening activity, tyrosinase inhibitory activity and melanin formation inhibitory activity were measured using B16F10 cells. In total 34 strains were obtained from various salted fish. The effective strains useful for the bioconversion process, showed that DPPH radical scavenging ability and polyphenol content were increased in the two kinds of microbial treatment groups compared to the untreated group. 16S rDNA sequencing analysis of the strains showed excellent in Pediococcus pentosaceus B1 in comparison. An increase of up-to 156% in anti-oxidative activity and 141% in polyphenol content was observed after bioconversion. In addition, after mixed fermentation the toxidty of Raw 264.7 and B16F10 cells tended to decrease and a significant increase was observed in anti-inflammatory activity as well. Also, tyrosinase activity and melanin significantly. synthesis decreased significantly.

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Inhibition of Leptin and Leptin Receptor Gene Expression by Silibinin-Curcumin Combination

  • Nejati-Koshki, Kazem;Akbarzadeh, Abolfazl;Pourhasan-Moghaddam, Mohammad;Abhari, Alireza;Dariushnejad, Hassan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6595-6599
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    • 2013
  • Leptin and its receptor are involved in breast carcinogenesis as mitogenic factors. Therefore, they could be considered as targets for breast cancer therapy. Expression of the leptin receptor gene could be modulated by leptin secretion. Silibinin and curcumin are herbal compounds with anti-cancer activity against breast cancer. The aim of this study was to assess their potential to inhibit of expression of the leptin gene and its receptor and leptin secretion. Cytotoxic effects of the two agents on combination on T47D breast cancer cells was investigated by MTT assay test after 24h treatment. With different concentrations the levels of leptin, leptin receptor genes expression were measured by reverse-transcription real-time PCR. Amount of secreted leptin in the culture medium was determined by ELISA. Data were statistically analyzed by one-way ANOVA test. The silibinin and curcumin combination inhibited growth of T47D cells in a dose dependent manner. There were also significant difference between control and treated cells in leptin expression and the quantity of secreted leptin with a relative decrease in leptin receptor expression. In conclusion, these herbal compounds inhibit the expression and secretion of leptin and it could probably be used as drug candidates for breast cancer therapy through leptin targeting in the future.

Comparative Study on the Content and Cytotoxicity of Pseudolaric Acid B in the Five Plant Parts of Pseudolarix kaempferi

  • Nugroho, Agung;Woo, Nam-Tae;Park, Kyoung-Sik;Kwon, Na-Yun;Jung, Woo-Nyung;Lee, Sang Kook;Kim, Dong-Hwa;Park, Hee-Juhn
    • Natural Product Sciences
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    • v.23 no.4
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    • pp.265-269
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    • 2017
  • Pseudolaric acids of Pseudolarix kaempferi (Pinaceae) have been known as diterpenoids with potent anti-fungal-, anti-microbial, and cytotoxic activities. In the present study, the five MeOH extracts were prepared from the five plant part (root bark, stem bark, leaf, the inner part of root, and cone) to find the relation between the concentration of pseudolaric acids and cytotoxicity. Pseudolaric acids B and C were isolated from the root bark of P. kaempferi to use them as standard compounds. The five extracts were tested on cytotoxicity against six cancer cell lines, A549 (lung), HCT116 (colon), MDA-MB-231 (breast), SNU638 (stomach), and SK-hep-1 (liver) by SRB assay, but against K562 (leukemia) by SRB- or MTT assay. HPLC quantification were performed on a Shisheido Capcell PAK C18 column ($5{\mu}m$, $4.6mm{\times}250mm$) using 254 nm wavelength. The cytotoxicity ($IC_{50}$, $0.36{\mu}g/ml$ on K562 cell lines) of the root bark extract was potent and the content (101.1 mg/g extract) of pseudolaric acid B was very high in the root bark. These results suggest that the MeOH extract obtained from the root bark could be developed as the anti-cancer agent with a high quantity of pseudolaric acid B.

Protective effects of remifentanil against H2O2-induced oxidative stress in human osteoblasts

  • Yoon, Ji-Young;Kim, Do-Wan;Kim, Eun-Jung;Park, Bong-Soo;Yoon, Ji-Uk;Kim, Hyung-Joon;Park, Jeong-Hoon
    • Journal of Dental Anesthesia and Pain Medicine
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    • v.16 no.4
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    • pp.263-271
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    • 2016
  • Background: Bone injury is common in many clinical situations, such as surgery or trauma. During surgery, excessive reactive oxygen species (ROS) production decreases the quality and quantity of osteoblasts. Remifentanil decreases ROS production, reducing oxidative stress and the inflammatory response. We investigated remifentanil's protective effects against $H_2O_2$-induced oxidative stress in osteoblasts. Methods: To investigate the effect of remifentanil on human fetal osteoblast (hFOB) cells, the cells were incubated with 1 ng/ml of remifentanil for 2 h before exposure to $H_2O_2$. For induction of oxidative stress, hFOB cells were then treated with $200{\mu}M$ $H_2O_2$ for 2 h. To evaluate the effect on autophagy, a separate group of cells were incubated with 1 mM 3-methyladenine (3-MA) before treatment with remifentanil and $H_2O_2$. Cell viability and apoptotic cell death were determined via MTT assay and Hoechst staining, respectively. Mineralized matrix formation was visualized using alizarin red S staining. Western blot analysis was used to determine the expression levels of bone-related genes. Results: Cell viability and mineralized matrix formation increased on remifentanil pretreatment before exposure to $H_2O_2$-induced oxidative stress. As determined via western blot analysis, remifentanil pretreatment increased the expression of bone-related genes (Col I, BMP-2, osterix, and $TGF-{\beta}$). However, pretreatment with 3-MA before exposure to remifentanil and $H_2O_2$ inhibited remifentanil's protective effects on hFOB cells during oxidative stress. Conclusions: We showed that remifentanil prevents oxidative damage in hFOB cells via a mechanism that may be highly related to autophagy. Further clinical studies are required to investigate its potential as a therapeutic agent.

Investigation of Nanofiber and Thermosensitive Scaffold for Intervertebral Disc through Organ Culture (기관배양을 통한 추간판 재생용 나노파이버 및 온도 감응성 지지체에 대한 검증)

  • Lee, Yong-Jae;Shin, Ji-Won;Shin, Ho-Jun;Kim, Chan-Hwan;Park, Ki-Dong;Bae, Jin-Woo;Seo, Hyoung-Yeon;Kim, Young-Jick;Shin, Jung-Woog
    • Journal of Biomedical Engineering Research
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    • v.28 no.4
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    • pp.512-519
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    • 2007
  • The purpose of this study is to investigate the potential of a novel tissue engineering approach to regenerate intervertebral disc. In this study, thermosensitive scaffold (chitosan-Pluronic hydrogel) and nanofiber were used to replace the nucleus pulposus (NP) and annulus fibrosus of a degenerated intervertebral disc, leading to an eventual regeneration of the disc using the minimally invasive surgical procedure and organ culture. In preliminary study, disc cells were seeded into the scaffolds and cellular responses were assessed by MTT assay and scanning electron microscopy (SEM). Based on these results, we could know that tissue engineered scaffolds might provide favorable environments for the regeneration of tissues. Organ culture was performed in fresh porcine spinal motion segments with endplates on both sides. These spinal motion segments were classified into three groups: control (Intact), injured NP (Defect), and inserting tissue engineered scaffolds (Insert). The specimens were cultivated for 7 days, subsequently structural stability, cell proliferation and morphological changes were evaluated by the relaxation time, quantity of DNA, GAG and histological examination. In these results, inserting group showed higher relaxation time, reduced decrement of DNA contents, and accumulated GAG amount. Consequently, the tissue engineered scaffolds used in this study seen to be a promising base scaffolds for regenerative intervertebral disc due to its capacity to absorb external dynamic loading and the possible ideal environment provided for disc cell growing.

Synthesis of Novel Pseudo-ceramide and Its Properties (신규 유사세라마이드의 합성과 그 특성)

  • Kim, Jin-Guk;Kim, Kyoung-Tae;Park, Sun-Hee;Lee, Bang-Yong;Kim, Ki-Ho;Kim, Young-Heui
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.34 no.1
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    • pp.43-50
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    • 2008
  • Ceramides, a constituent of stratum corneum lipids, play a crucial role in the formation and maintenance of the epidermal permeability barrier. As in many other skin disorders, atopic dermatitis and psoriasis show decrease and transformation of the ceramides. The application of ceramide has been demonstrated to be efficient in the repair of these skin disorders. Nevertheless, natural ceramides are still too expensive and small in quantity to be used as a cosmetic ingredient. Although a lot of pseudo-ceramides have been developed and on the market until now, those pseudo-ceramides did not fully meet the consumer's needs, therefore, there is still a demand for a novel pseudo-ceramides. We synthesized a novel pseudo-ceramide BPC-16 from 2-(2-amino-ethylamino)-ethanol(AEEA), which was characterized by structures having both amide bonds and hydroxyl groups as hydrophilic units, as well as two long alkyl chains. We formulated emulsion with BPC-16, cholesterol, stearic acid, and other components to make an emulsion. These emulsion showed a typical optical anisotropy on cross-polarized microscopy. This 'Maltese cross' appearance is a characteristic figure observed in concentric lamellar emulsion under cross-polarized microscopy. In cytotoxicity assay using MTT in monolayer and three dimension(3D) cell culture, a BPC-16 showed only negligible cytotoxicity up to the effective concentration for barrier repair and moisturization(less than 10 mM). In the measurement of TEWL, this BPC-16 showed significant recovery of water-retaining properties when it was topically applied to either SDS-induced dry skin or normal skin compared to that of base cream. This novel pseudo-ceramide BPC-16 showed as effective in skin barrier repair and moisturization as natural ceramides.