• 생명과학회지
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• 제8권1호
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• pp.67-71
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• 1998

다량의 시료를 빠르게, 적은 격비로, 간다히 검색할수 있는 예비선별법을 찾던중, ccDNA와 sulforhodamine B를 각각 사용한 미생물배양액내의 신규항암후보물질의 탐색을 시도하였다. cccDNA법으로는 3.3%가 positive 반응을 나태내었으며, SRB assat에서는 A549와 SK-OV-3에 활성을 나타내는 4개의 발효여액을 확인한수 있었다.

• 한국식품영양과학회지
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• 제28권1호
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• pp.240-245
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• 1999

Growth inhibitory effect of doenjang(Korean soypaste) methanol extracts in SRB assay using AGS human gastric adenocarcinoma cell, Hep 3B human hepatocellular carcinoma cell and HT 29 human colon cancer cell was studied. The treatment of doenjang methanol extracts(2mg/assay) to the AGS, Hep 3B and HT 29 cancer cells inhibited the growth of the cancer cells by 55%, 60%, and 71%, respectively. Doenjang methanol extracts exhibited the highest inhibitory effect among other soybean fermented foods and original materials in the SRB assay. In addition, to separate active compounds of doenjang methanol extracts, we fractionated the doenjang with hexane, methanol, dichloromethane, ethylacetate and butanol. Growth inhibitory effect on the AGS, Hep 3B, HT 29 and MG 63 cancer cells was the highest in the fractions of dichloromethane and ethylacetate among other solvent fractions of the doenjang. These results showed that some compounds contained in the fractions of dichloromethane and ethylacetate might play a role on the anticanceric effect of doenjang.

• Journal of Microbiology and Biotechnology
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• 제14권2호
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• pp.350-355
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• 2004

Sulforhodamine B (SRB) assay is a rapid, sensitive, and inexpensive method for measuring cell proliferation and chemosensitivity. However, the lactate dehydrogenase (LDH) release assay is generally used to measure cytototoxicity of infectious microorganisms against host cells. In this study, we investigated the possibility of applying the SRB assay to determine cytotoxicity for infectious microorganisms, and compared the results with those obtained by the LDH release assay. We used Vibrio vulnificus as a model of infectious microorganisms. The SRB assay showed that V vulnificus strongly induced cytotoxic activity against human intestinal cells, Caco-2 and INT-407 cells. The degree of cytotoxicity closely correlated with infection time and number ratios of V. vulnificus to intestinal cells (MOI, multiplicity of infection). Furthermore, cytotoxicity values obtained by SRB assay correlated well with results obtained by the LDH release assay, and both assays gave a linear response with respect to MOI Heat-inactivation of V. vulnificus for 35 min at $60^{\circ}C$ did not induce cytotoxic activity, indicating that viability of V. vulnificus is crucial for cytotoxic activity against intestinal cells. Although both assays are suitable as cytotoxicity endpoints, the SRB assay is recommended for measuring cytotoxicity of infectious microorganisms against host cells because of its significantly lower cost and more stable endpoint than the LDH release assay.

• Journal of Applied Biological Chemistry
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• 제43권1호
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• pp.59-61
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• 2000

The cytotoxic activities of the methanol extracts of 44 plant species in 31 families against five human solid A549 (lung), SK-OV-2 (ovarian), SK-MEL-2 (melanoma), XF-498 (central nervous system), and HCT-15 (colon) tumor cell lines were examined using the sulforhodamine B (SRB) assay. Responses varied with both cell line and plant species used. Potent cytotoxic activities ($ED_{50}$, <$40{\mu}g/ml$) against all model tumor cell lines were produced from the extracts of Rhus chinensis gall (Galla rhois), Betula platyphylla var. japonica bark, Inula helenium root, Cinnamomum cassia bark, Cinnamomum sieboldii root bark, Lysimachia davurica whole plant, and Evodia rutaecarpa fruit. These plants may be useful for developing new types of naturally occurring anti-tumor agents.

• 동의신경정신과학회지
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• 제11권1호
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• pp.1-18
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• 2000

To study the effects of Ramulus et Uncus Uncariae (REUU) on oxygen free radical-mediated damage by hydrogen peroxide $(H_{2}O_{2})$ on cultured spinal sensory neurons, in vitro assays such as MTT assay, NR assay, neurofilament enzymeimmuno assay (EIA), sulforhodamine B (SRB) assay, assay for lactate dehydrogenase (LDH) activity and assay for lipid peroxidation were used in cultured spinal dorsal root ganglion neurons derived from mice, Spinal dorsal root ganglion neurons were cultured in media containing various concentrations of $H_{2}O_{2}$ for 5 hours, after which the neurotoxic effect of $H_{2}O_{2}$ was measured by in vitro assay. The protective effect of the herb extract, Ramulus et Uncus Uncariae (REUU) against H2O2-induced neurotoxicity was also examined. The results are as follows. 1. In NR assay and MTT assay, $H_{2}O_{2}$ significantly decreased the cell viability of cultured mouse spinal dorsal root ganglion neurons according to exposure concentration in these cultures. An additional time course study was done on these cultures. 2. Cultured spinal dorsal root ganglion neurons which were exposed to various concentrations of $H_{2}O_{2}$ showed a quantitative decrease of neuronal cells by EIA and of total protein by sulforhodamine B (SRB) assay, while they showed an increase of both lipid peroxidation and LDH activity. 3. The effect of Ramulus et Uncus Uncariae (REUU) on $H_{2}O_{2}$ induced neurotoxicity showed a quantitative increase in both neurofilament and total protein, but showed a decrease of lipid peroxidation and LDH activity. These results suggest that $H_{2}O_{2}$ has a neurotoxic effect on cultured spinal dorsal root ganglion neurons from mice and that the herb extract, Ramulus et Uncus Uncariae (REUU), was very effective in protecting $H_{2}O_{2}$ induced neurotoxicity by decreasing lipid peroxidation and LDH activity.

• 생명과학회지
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• 제17권1호
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• pp.76-81
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• 2007

메주의 헥산 및 메탄올 추출물과 그것을 용매로 분획하여 얻어진 디클로로메탄, 에틸아세테이트, 부탄올, 물 분획물들에 의한 SOS chromotest 실험계에서 항돌연변이 효과 및 SRB assay를 이용한 인체 암세포들의 성장 억제 효과에 대하여 검토하였다. SOS chromotest 실험계의 경우, 첨가농도 $100{\mu}g/assay$에서 메주의 디클로로메탄 및 에틸아세테이트 분획물은 각각 91%, 91%로 MNNG에 대하여 강한 항돌연변이 효과를 나타났다. AGS 인체 위암세포와 Hep 3B 인체 간암세포를 이용하여 항암효과를 실험한 결과, 메주 에틸아세테이트분획물은 첨가농도 $200{\mu}g/assay$에서 각각 93% 및 91%로 암세포 성장을 크게 저해시켰고 그 저해 효과는 디클롤메탄 분획물보다 더 높았다. 인체 위암세포인 AGS를 이용한 SRB assay을 이용한 억제 효과 실험에서 메주의 에틸아세테이트 분획물은 첨가농도 2 mg/assay에서 인체 위암 및 간암세포의 증식을 각각 64% 및 71%로 억제하여 분회물들 중 가장 높은 저해효과를 보였고 헥산 및 디클로로메탄 분획물은 그 다음으로 억제 효과가 높았다. 이러한 결과는 된장 메탄올 추출물과 그 분획물의 결과와 유사한 경향을 보였으므로 메주와 된장이 여러 종류의 미생물, 곰팡이류와 세균류들에 의해 발효과정을 거치는 동안 원재료인 콩에서는 없었던 혹은 함량이 적은 성분들이 생성되거나 증가되어 항암효과를 나타내는 것으로 추정되어 진다.

• 생명과학회지
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• 제17권10호
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• pp.1368-1373
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• 2007

대두를 먼저 헥산(Hex)으로 지방을 제거한 후 메탄올로 추출한 추출물에서 항돌연변이성 및 항암활성 물질을 분리하기 위해 극성이 다른 용매인 디클로로메탄($CH_2Cl_2$), 에틸아세테이트(EtOAC), 부탄올(BuOH)로 분획하여 각 분획물의 항돌연변이 및 항암활성 효과를 검토하였다. $AFB_1$에 대한 돌연변이 억제효과는 대두의 메탄을 추출물(MeOH)의 분획물들 중 디클로로메탄 분획물($CH_2Cl_2$)과 에틸아세테이트 분획물(EtOAc)은 2.5 mg/plate 첨가농도에서 각각 83%로 상당히 높은 항돌연변이 효과를 나타내었다. 중간 분획물(Inter)의 경우는 56%의 항돌연변이 효과를 나타내었으나 다른 분획물들의 경우 항돌연변이 효과가 낮았다. 한편 MNNG의 경우, $AFB_1$에 비해 돌연변이 저해효과가 떨어지지만 그 중에서도 에틸아세테이트 분획물(EtOAc)이 67%로 가장 높은 항돌연변이 효과를 나타내었으며 중간 분획물과 디클로로메탄 분획물($CH_2Cl_2$)은 각각 63%, 49%의 돌연변이 저해효과를 나타내었다. 대두의 메탄을 추출물은 첨가농도 2 mg/assay에서 44%의 인체 위암세포의 증식 억제효과를 가졌고 대두 메탄을 추출물의 분획물들 중 에틸아세테이트층(EtOAC)의 저해 효과가 가장 높았는데 동일농도에서 66%로 위암세포 성장을 저해시키는 효과를 나타내었고 디클로로메탄($CH_2Cl_2$)과 중간 분획물(Inter)의 경우에는 각각 54% 및 51%의 저해효과를 나타내었다. Hep 3B 인체 간암세포의 경우, 대두 메탄을 추출물을 2 mg/assay 투여했을 때 60%의 저해효과를 나타내었으며 여기에서도 분획물들 중에서 에틸아세테이트 분획물(EtOAC)이 73%로 가장 높은 저해효과를 보였다. 대두의 메탄을 추출물은 첨가농도 2 mg/assay에서 44%의 인체 결장암세포 증식억제효과를 가졌고 대두 메탄을 추출물의 분획물들 중 에틸아세테이트층(EtOAC)의 저해 효과가 가장 높았는데 동일농도에서 77%의 결장암세포의 성장을 저해시키는 효과를 나타내었다. 이상의 결과로부터 대두 분획물들 중 특히 에틸아세테이트 분획물에 의한 항돌연변이 및 항암활성효과가 가장 높았으므로 이 분획물에 주로 isofavone류가 함유되어 있으므로 에틸아세테이트 분획물을 더욱 정제하여 연구할 필요가 있다고 여겨진다.

• Toxicological Research
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• 제16권2호
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• pp.101-107
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• 2000

Cannabidiol derivatives (1, 2 and 3), 5-fluorouracil (4, 5-FU) and adriamycin (5, AM) were tested for their growth inhibitory effects against human tumor cell lines using two different 3-{4,5-dimeth-ylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and sulforhodamine B protein (SRB) assay. The light microscopic study showed morphological changes of the treated cells. Disruptions in cell organelles were determined by colorimetric methods; MTT assay and STB assay. These results suggest that cannabidiol (1, CBD) retains the most growth-inhibitory activity against human tumor cell lines.

• 약학회지
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• 제42권3호
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• pp.307-311
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• 1998

This study was carried out to evaluate antitoxic effects Taraxaci Herba extract against Cadium by calorimetric methods. The antitoxic activity of Taraxaci Herba ex tract in NIH 3T3 fibroblasts was evaluated by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-phenyl-2H-tetrazoliumbromide), NR (Neutral red) and SRB (Sulforhodamine B protein) assay. The light microscopic study was carried out to observe morphological changes of the treated cells. These results were obtained as follows; The concentration of $10^{-2}mg/ml$ of Taraxaci Herba extract was shown significant antitoxic activity. The number of NIH 3T3 fibroblasts were antitoxic and tend to regenerate. These results suggest that Taraxaci Herba extract retains a potential antitoxic activity.

• Archives of Pharmacal Research
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• 제21권3호
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• pp.353-356
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• 1998

Geraniol (1), olivetol (2), cannabinoids (3 and 4) and 5-fluorouracil (5) were tested for their growth inhibitory effects against human oral epitheloid carcinoma cell lines (KB) and NIH 3T3 fibrobalsts using two different 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and sulforhodamine B protein (SRB) assay. Cannabigerol (3) exhibited the highest growth-inhibitory activity against the cancer cell lines.