• Title/Summary/Keyword: MTT 실험

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Physico-chemical Properties and Antibacterial Activities of Lactonic Sophorolipid (락톤형 소포로리피드의 물리화학적 특성 및 항균효과)

  • Cho, Soo A;Eom, Gyeong Tae;Jin, Byung Suk
    • Applied Chemistry for Engineering
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    • v.30 no.3
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    • pp.303-307
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    • 2019
  • Sophorolipid is a biological surfactant of the glycolipid structure produced by Candida bombicola, which generally exists as a mixture of acidic and lactonic forms. In this study, we investigated physico-chemical properties, antibacterial activities, and cytotoxicity of the sophorolipid containing more than 96% of the lactonic form, produced by the gene regulation of production strains and application of a metabolic engineering technique. The lactonic sophorolipid showed a weak acidity in the range of pH 3.2~4.6 when diluted in water at the concentrations from 1 to 0.001 wt%. The $pK_a$ value of the lactonic sophorolipid was estimated to be around 4.3 from the acid-base titration curve. The critical micelle concentration (CMC) of the lactonic sophorolipid was $10^{-2}wt%$, at which the surface tension of aqueous solution was reduced to 36 mN/m. The lactonic sophorolipid showed the minimum inhibitory concentrations (MIC) of $1{\times}10^{-3}$ and $5{\times}10^{-3}g/mL$ against Propionibacterium acnes and Corynebacterium xerosis, respectively. The MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] assay showed that cytotoxicity of the lactonic sophorolipid was ten times lower than that of triclosan.

Analysis of Physiological Activity and Cytotoxicity of Residue Extracts after Pepper Harvest (고추 수확 후 잔재물 추출물의 생리활성과 세포독성 분석)

  • Kim, Tae-Won;Jeon, Byeong-Gyun;Lee, Sung-Ho
    • Journal of Life Science
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    • v.30 no.12
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    • pp.1085-1091
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    • 2020
  • The purpose of this study was to investigate the value of residue discarded after harvesting a species of pepper (Capsicum annuum L.). This study analyzed the physiological activity and cytotoxicity of a fermented extract and a hot water extract gathered from the residues of the Cheongyang pepper and the Nokkwang pepper. The total nitrogen, phosphoric acid, potassium, calcium, and magnesium contents of the fermented extract of the Nokkwang pepper were approximately twice as high as those of the Cheongyang pepper, while the hot water extracts had similar levels of these components across both peppers. Among the microelements, only boric acid, iron, and silicon components were detected, whereas zinc, manganese, molybdenum, and copper components were not detected in either extract. The total polyphenol and flavonoid contents of the fermented extracts were more than two times higher than those of the hot water extracts for both peppers. The DPPH radical scavenging ability (indicating antioxidant activity) of the fermented extract was higher than that of the hot water extract, while the ABTS radical scavenging ability (indicating antioxidant activity) of the hot water extract was higher than that of the fermented extract. A cytotoxicity test was conducted on the extracts using an MTT assay. Both extracts exhibited weak cytotoxicity at all concentrations for both peppers. These results suggest that the extract of residue discarded after harvesting these peppers could be suitable for functional feed or agricultural materials.

Anti-cancer and Anti-microbial Effect of the Fraction Isolated from Pyrus ussuriensis Leaves (산돌배나무(Pyrus ussuriensis) 잎 분획물의 항암 및 항균활성에 관한 연구)

  • Lee, Chang-Eon;Kim, Young-Hun;Lee, Byung-Guen;Lee, Do-Hyung
    • Journal of Korean Society of Forest Science
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    • v.100 no.2
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    • pp.136-141
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    • 2011
  • This study was conducted to confirm the application as ingredients of cosmetics through an examination of the function for anti-cancer and anti-microbial of the fraction isolated from Pyrus ussuriensis leaves. The dried leaf of P. ussuriensis were extracted with acetone-$H_{2}O$ (6:4, v/v), concentrated and fractionated with the upper layer of acetone on a separatory funnel. Each fraction was freeze dried, then a portion of acetone soluble powder was chromatographed on a Sephadex LH-20 column using a series of aqueous methanol as eluents and also used the MIC-gel using a series of aqueous methanol as developing solvent. The isolated compounds were identified by silica-gel TLC. The growth inhibition activity was measured using the MTT assay by the mouse meltioma (B16F10) cell. The cancer cell growth inhibition rate of fractions isolated from P. ussuriensis leaf was 80%. In anti-microbial activity test, the fraction of P. ussuriensis with 0.25 mg/disc resulted in the clear zone of 1.3 cm and 2 cm for Staphylococcus aureus and S. epidermidis of gram positive bacillus, respectively. In Escherichia coli of gram negative bacillus, the fraction with 0.5 mg/disc resulted in the clear zone of 1.1 cm~1.5 cm each fraction. From these results, we confirmed that acetate fraction of P. ussuriensis has a great potential as a natural ingredients with a anti-cancer and anti-microbial source.

Whitening Effect of Abelmoschus esculentus on Melanoma Cells (B16F10) (B16F10 세포에서의 오크라 추출물의 미백 활성 검증)

  • Yoo, Dan-Hee;Lee, In-Chul
    • Microbiology and Biotechnology Letters
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    • v.49 no.4
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    • pp.485-492
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    • 2021
  • In this study, the whitening effect of Abelmoschus esculentus extract was investigated to confirm its applicability in cosmetics. To determine the whitening effect, the tyrosinase-inhibitory activity of Abelmoschus esculentus hot water extract (AEWE) and 70% ethanol extract (AEEE) was measured. At the final concentration of 1000 ㎍/ml, AEWE showed an inhibitory activity of 22.2% and AEEE of 32.8%. To determine the whitening effect at the cellular level, the viability of melanoma cells treated with AEWE and AEEE was evaluated using the MTT assay. At concentrations of 100 ㎍/ml or less, both AEWE- and AEEE-treated groups showed cell survival rates of >95%. Furthermore, in both AEWE- and AEEE-treated melanoma cells, the melanin content decreased in a concentration-dependent manner. The inhibitory effects of AEWE and AEEE used at 5, 10, 50, and 100 ㎍/ml on protein expression were measured by western blot, with β-actin as the positive control. At a concentration of 100 ㎍/ml, AEWE showed an inhibitory effect of 88.1%, 24.8%, 62.2%, and 42.9% on microphthalmia-associated transcription factors (MITF), tyrosinase, tyrosinase-related proteins (TRP)-1, and TRP-2 factors, respectively. At the same concentration, AEEE showed inhibitory effect of 65.3%, 58.3%, 66.2%, and 65.3% against MITF, tyrosinase, TRP-1, and TRP-2 factors, respectively. In conclusion, the whitening effects of AEWE and AEEE were verified, and their applicability as a natural ingredient in cosmetics was confirmed.

Verification of Whitening Activity of Inonotus obliquus Extracts in B16F10 Melanoma Cells (차가버섯(Inonotus obliquus) 추출물의 B16F10 멜라노마 세포에서의 미백활성 검증)

  • Song-Yoon Choi;Je-Back Lee;Hyeon-Ji Yeom;Min-Jeong Oh;Jin-Young Lee
    • Journal of Life Science
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    • v.34 no.2
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    • pp.105-112
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    • 2024
  • In this study, the various whitening activities of Inonotus obliquus were assessed for potential use as functional cosmetic materials. When measuring electron donating ability to confirm the antioxidant ability of I. obliquus extract, increased activity was observed as the concentration increased, and it was found an outstanding antioxidant capacity of 82.1% at a 1,000 ㎍/ml concentration. Also, the tyrosinase inhibitory effect, related to a whitening effect, was found to have inhibitory activity that increased in a concentration-dependent manner. The results of verifying the viability of melanoma cells (B16F10) using an MTT assay showed cell viability of more than 80% at concentrations below 100 ㎍/ml. Therefore, cell-related experiments were conducted at 25, 50, and 100 ㎍/ml concentrations. By measuring protein expression related to melanin synthesis via treating B16F10 cells with I. obliquus extract, it was confirmed that protein expression was inhibited in all factors, depending on the concentration. TRP-1 and MITF appeared by 40.1% and 64.2% amount of expression, respectively, at 100 ㎍/ml concentrations, and tyrosinase and TRP-2 were verified as having better protein expression inhibition than arbutin. In measuring mRNA expression related to melanin synthesis by treating B16F10 cells with I. obliquus extract, it was confirmed that mRNA expression was suppressed as the concentration increased. Accordingly, it was confirmed that I. obliquus extract has excellent whitening activity and could be used as a cosmetic material.

Enhancement of Anticarcinogenic Effect by Combination of Lycii fructus with Vitamin C (구기자 추출성분의 항발암 효과 및 비타민 C첨가에 의한 상승효과)

  • 박윤자;김미향;배송자
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.1
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    • pp.143-148
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    • 2002
  • We investigated the cytotoxicity effects of Lycii fructus (LF) on HePG2, MCF7 and C6 cell lines by the MTT assay. We extracted the methanol (LFM) and fractionated to five partition layers. Among partition layers, the ethylether partition layer (LFMEE) was showed the strongest cytotoxic effects on all cancer cell lines. The hexane partition layer (LFMH) also was showed significant cytotoxic activities on Hela and MCF-7 cell lines. We also determined the induction of intracellular quinone reductase (QR) activity on HepG2 cells. Among various partition layers of Lycii fructus; LFMH was showed the most effective such induced effect such as 1.85 to the control value of 1.0. And we also determined the enhancement of anticarcinogenic effect by combination of Lycii fructus with vitamin C on all cell lines. These results suggest that potentially useful anticarcinegenic chemicals could be isolated from LFMEE and LFMH of the Lycii frutus and also we found the enhanced effect by the combination of various partition layers of LFM with vitamin C.

Collagen synthesis ability and inhibitory effect of MMPs in keratinocytes of Lysimachia christinae Hance Extract (금전초 추출물의 케라티노사이트 내 collagen 합성능 및 MMPs 억제효과)

  • Kim, Ju-Eun;Choi, Yun-Sik;Kim, Hye Kyung;Jang, Young-Ah
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.4
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    • pp.820-829
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    • 2020
  • This study was conducted to investigate the anti-oxydant and anti-winkle efficacy as cosmetics ingredient of Lysimachia christinae Hance. Recently, the study of wrinkle improvement of natural products has received continuous interest. so we looked at relationship between reactive oxygen species (ROS) generation and pro-collagen synthesis and matrix metalloproteinases (MMPs) through this study. L. christinae Hance were extracted with 70% ethanol (LcHE) and distilled water (LcHW), respectively, and the experiment was conducted. LcHE had better ROS inhibition effect than LcHW and showed no toxicity up to 250 ㎍/mL concentration as a result of MTT assay in HaCaT cells, so we selected LcHE and conducted the wrinkle improvement material study. We confirmed that the synthesis of type-1 pro-collagen reduced by UVB is activated through pro-collagen synthesis assay. we confirmed that LcHE inhibited the increase in MMP-1 -3 -9 of MMPs induced by UVB in skin cells through western blot and we also performed real-time PCR to confirm the effect of the extract with dependence of concentration at mRNA levels. Therefore, it is expected that Lysimachia christinae Hance is used as a natural material for cosmetics that can effectively prevent wrinkles and skin aging by UVB.

Bulnesia Sarmienti Aqueous Extract Inhibits Inflammation in LPS-Stimulated RAW 264.7 Cells (RAW 264.7세포에서 lipopolysaccharide로 유발시킨 염증반응에 대한 Bulnesia sarmienti 열수추출물의 억제효과)

  • Cheon, Yong-Pil;Mollah, Mohammad Lalmoddin;Park, Chang-Ho;Hong, Joo-Heon;Lee, Gee-Dong;Song, Jae-Chan;Kim, Kil-Soo
    • Journal of Life Science
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    • v.19 no.4
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    • pp.479-485
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    • 2009
  • Bulnesia sarmienti (BS), a traditional South American herbal medicine native to Gran Chaco, has been used to treat various human ailments. We investigated the cytotoxic activities and the inhibitory effects of BS bark extract(0, 50, 100 and $200\;{\mu}g/\;mL$) on the production of nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), cyclooxygenase (COX) and proinflammatory cytokines ($IL-1{\beta}$, IL-6 and $TNF-{\alpha}$) in the lipopolysaccharide (LPS) (100 ng/ml)-stimulated murine macrophage cell line RAW264.7. The levels of NO, COX, PGE2 production and proinflammatory cytokines ($IL-1{\beta}$, IL-6 and $TNF-{\alpha}$) were measured by ELISA kit. Cell viability, as measured by the MTT assay, showed that BS extract had no significant cytotoxicity in RAW264.7 cells. BS extract significantly inhibited the LPS-induced NO, $PGE_2$ and COX production accompanied by an attenuation of $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ formation in macrophages. These results suggest that BS extract has potential as an herbal medicine for the treatment of inflammatory diseases.

Estimation of Anti-proliferative Activity of Saccharin against Various Cancer Cell Lines and MSCs (다양한 암세포 주와 MSCs에 대한 Saccharin의 항증식성 평가)

  • Choi, Jeong Su;Park, Sang Yong;Yang, Man Gil;Lee, Dong Beom;Lee, Tae Bok;Heo, Ji Hye;Lee, Min Woo;Kim, Suhng Wook
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.3
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    • pp.169-175
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    • 2016
  • Saccharin (o-benzoic sulfimide) is the first artificial and non-caloric sweetener that was first synthesized in 1879. In this study, we examined the biological activity of saccharin against various human cancer cell lines and human bone marrow-derived mesenchymal stem cells. A viability assay based on the conversion of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was performed to test for the cytotoxicity of saccharin about the four human cancer cell lines (H460, H157, A549 and SKOV3), one murine cancer cellline (Raw264.7), and MSCs. In order to find the differentially expressed gene in saccharin-treated MSCs against untreated MSCs, we performed annealing control primer (ACP)-based differential display reverse transcriptionp-olymerase chain reaction (DDRT-PCR). All tested cells were treated with saccharin at various concentrations (0.0, 4.8, 7.2, 9.6, 12.0, 14.4 mg/mL) for 48 hr. The number of metabolically active cancer cells decreased when treated with the saccharin at various concentrations for 48 hr as compared with the untreated cells. The decrease in cell survival was more evident with increasing concentrations of saccharin. Moreover, novel candidate genes, which were differentially expressed in MSCs in response to saccharin, were identified in 16 bands on 2% agarose gel. This revealed 16-7 up-regulated and 9 down-regulated-differentially expressed genes indicated by arrows. One of these candidate genes was a FK506-binding protein gene. The functional roles of FK506 binding proteins, with respect to the activities of stem cell proliferation, were not characterized. Further studies are required to get a better understanding of FK506-binding proteins in its roles in increasing stem cell proliferative activities from using saccharin.

Study on Anti-inflammatory and Anti-microbial Effect of Pinus rigida Mill. inner Bark Extracts as a Cosmetic Material (리기다소나무(Pinus rigida Mill.) 내수피 추출물에 대한 화장품 소재로써의 항염 및 항균효과)

  • Jang, Min-Jung;Kim, Young-Hun;An, Bong-Jeun;Lee, Chang-Eeon;Lee, Jin-Tae;Kim, Sea-Hyun;Lee, Byung-Guen;Lee, Do-Hyung
    • Journal of Korean Society of Forest Science
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    • v.97 no.3
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    • pp.215-220
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    • 2008
  • Recently, there has been a great deal of interest in the applications of plant-based extracts to both cosmetic and medicinal industries. The objective of this study was to investigate the anti-inflammatory and antimicrobial effect of P. rigida extracts by water and ethyl acetate. Anti-inflammatory and anti-microbial effect of P. rigida extracts by water and EtOAc were investigated by using nitrite scavenging ability, nitric oxide production and anti-microbial ability. In the test of nitrite scavenging ability, P. rigida extracts by water and EtOAc showed 88.7% and 99% at 100 ppm concentration, respectively. The cell viability was measured using the MTT assay at 24 hours after P. rigida extracts as shown in over 80%. Anti-inflammatory effect was examined in LPS stimulated RAW 264.7 cells. NO productions in LPS and P. rigida extracts stimulated group were decreased in a concentration and were dependent on time as compared with LPS stimulated. The water extracts showed the highest inhibition at the 100 ppm concentration. In anti-microbial activity test, the water extract with 3.0 mg/disc resulted in the clear zone of 14 mm, and ethyl acetate with that of 15 mm for Staphylococcus aureus. However, P. rigida extracts didn't show any growth inhibitory effect on Esherichia coli. These results indicate that the extracts of P. rigida have anti-inflammatory activities as a cosmeceuticals.