• Toxicological Research
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• 제16권1호
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• pp.77-82
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• 2000

This study was conducted to assess a possible alternative method as replacements for in vivo test. The human fibroblasts were exposed to several photoxic chemicals (promethazine, neutral red, chlortetracyclone, amiodatone, bithional, 8-methyooxypsorale) and non-phototoxi substance, ammonium laureth sulfate and irradiatied with 5 J/$cm^2$ of UVA (3320~420nm). The cell viability was measured by NRU (neutral red uptake) assay. The photoxic potential of test chemicals in the NRU PT (phototoxicity test) was assessed by determining the PIF (photoirritancy Factor) by using a cut-off value of 5. The NRU PT responses of most chemicals showed a close agreement with in vivo response except bithinol. There was a relatively good agreement between in vitro NRU assay and in vivo data. These results suggest that NRU assay using fibroblast could be used to predict the phototoxicity.

• 대한화장품학회지
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• 제35권3호
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• pp.193-202
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• 2009

본 연구에서는 in vivo 광독성 시험을 대체할 수 있는 방법에 관한 연구를 수행하였다. 인체 유래의 섬유아세포를 활용하여 광독성 물질(promethazine, chlorpromazine chlortetracycline, 8-methoxypsoralen, neutral red, bithionol)과 비광독성 물질(cinnamic aldehyde, p-aminobenzoic acid, sodium lauryl sulfate, L-cysteine)을 평가하였다. 세포 생존율은 neutral red uptake (NRU)로 평가하였다. NRU 광독성 시험 결과 bithionol를 제외한 화합물에서 모두 in vivo 실험결과와 유사한 결과를 보였다. 같은 방법으로 화장품 성분인 $Medimin^{(R)}$ A, $Medimin^{(R)}$ D, $LG^{(R)}$ 106W, $Phytoclear^{(R)}$ EL-1, 단삼동 추출물, 미인초 추출물, 산거울 추출물, $Parsol^{(R)}$ MCX와 $Parsol^{(R)}$ 1789를 평가 하였다. 평가 결과 단삼동 추출물을 제외한 원료에서 광독성이 없는 것으로 평가되었다.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2001년도 International Symposium on Signal transduction in Toxicology
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• pp.109-109
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• 2001

In order to evaluate the neutral red uptake assay as an alternative method for phototoxicity test, we compared the potential of phototoxicity in vitro in cultured human fibroblasts and 3T3 fibroblast cells derived from Balb/c mice. Both fibroblasts were exposed to various known phototoxic chemicals (promethazine, neutral red, chlorpromazine, chlortetracycline, amiodarone, bithionol, 8-methoxypsoralen) and non-phototoxic chemical (ammonium laureth sulfate) and irradiated with 5 J/cm$^2$ of UVA.(omitted)

• 농약과학회지
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• 제18권1호
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• pp.8-13
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• 2014

본 연구는 5가지 제품농약을 이용하여 어류 급성독성시험 결과 (반수치사농도)와 잉어의 표피에서 유래된 EPC 세포를 이용한 neutral red uptake 결과 (반수저해농도)를 비교함으로써 동물 실험의 대체 가능성을 평가하기 위하여 수행되었다. 어류 급성 독성시험은 왜몰개 (Aphyocypris chinensis)를 포함하여 OECD와 농촌진흥청의 농약에 대한 독성시험기준에서 추천하는 어종인 송사리 (Oryzias latipes)와 잉어 (Cyprinus carpio)를 이용하여 수행하였다. 5가지 제품 농약에 대한 민감도는 어류에 비하여 세포에서 약 10배 더 낮게 확인되었지만, 독성을 서열화 하였을 때 나타나는 순서는 두 가지 방식에서 모두 비슷하게 나타났다. 5가지 제품 농약에 대한 세포와 어류 독성값의 상관성을 분석한 결과는 A. chinensis, O. latipes와 C. carpio에서 각각 $r^2=0.38$ (p = 0.26), $r^2=0.76$ (p = 0.05), $r^2=0.90$ (p = 0.01)을 나타내었다. 본 시험의 결과, EPC 세포를 이용한 NRU 시험은 O. latipes와C. carpio에 대한 어류 독성시험 결과와 상관성이 높으므로 향후 더 많은 약제시험을 통해 어류 급성독성시험의 대체시험법으로서의 가능성이 기대된다.

• 생약학회지
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• 제33권1호통권128호
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• pp.13-17
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• 2002

Helicobacter pylori infection is associated with type B gastritis, peptic ulcer, and gastric cancer. The vacuolation of cells induced by H. pylori is thought to be essential for the initiation and maintenance of gastric infection. The roles of H. pylori cytotoxin, urease, and ammonia in the vacuolation of HeLa cells were determined. Ammonium chloride augmented the neutral red uptake induced by H. pylori toxin. Acetohydroxamic acid (AHA) failed to block the neutral red uptake induced by H. pylori toxin. Leweifang significantly prevented the vacuolation of HeLa cells induced by H. pylori toxin or H. pylori toxin and ammonium chloride. Further investigation is required to determine the mechanisms of Leweifang for the inhibition of vacuole formation of eukaryotic cells in response to the H. pylori toxin.

• Journal of Pharmaceutical Investigation
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• 제25권4호
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• pp.365-369
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• 1995

The acute cytotoxicities of chloroquine sulfate, propranolol, ascorbic acid, acetylsalicylic acid and acrylamide on cultured adult rat hepatocytes were evaluated by the use of LDH leakage and NR uptake test. On the basis of $IC_{50}$ values, the rank order of cytotoxicities of these drugs in both tests was chloroquine sulfate > propranolol > acetylsalicylic acid > ascorbic acid. The $IC_{50}$ of LDH test was very similar to that of NR uptake test. Thus, we concluded that both tests are reliable and sensitive methods in detecting toxicity in adult cultured rat hepatocytes.

• 한국연초학회지
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• 제29권2호
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• pp.110-117
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• 2007

In vitro toxicity tests such as cytotoxicity, mutagenicity and genotoxicity assay are useful for evaluating the relative toxicity of smoke or smoke condensates obtained from different cigarette configurations. A major disadvantage of these tests is relatively time-consuming, complicated and expensive. Recently, a cell-free glutathione consumption assay (GCA) as a rapid and simple screening method for the toxicity assessment of smoke has been reported by Cahours et al. (CORESTA, 2006). This study was carried out to assess the GCA application capable of predicting the toxicity of gas/vapor phase (GVP) of cigarette smoke and to identify individual compounds responsible for the glutathione (GSH) consumption in smoke. Each GVPs from 2R4F, standard cigarette, carbon filter cigarette (ExC) and new carbon filter cigarette (ExN), test cigarettes were collected by automatic smoking machine and evaluated the relative toxicity by GCA and neutral red uptake (NRU) assay. Toxic compounds existed in smoke were also chosen, relative toxicities of these compounds were screened by using two methods and compared individually. The overall order of toxicity by GCA was 2R4F > ExC > ExN, which was consistent with the result of Neutral Red Uptake assay. The levels of carbonyl compounds of ExN were lower than those of 2R4F and ExC, indicating that GSH consumption was associated with carbonyl compound yields. A major toxicant under current study is acrolein, which contributed to more than half of the GSH consumption. Collectively, the toxicity of GVP determined by GCA method may be mainly attributed to acrolein.

• Toxicological Research
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• 제18권3호
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• pp.227-232
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• 2002

In order to find out the appropriate in vitro method for high correlation with in vivo, we com-pared the sensitivities of phototoxicity (PT) in vitro method between in human keratinocytes, HaCaT cells and in 3T3 fibroblast cells derived from Balb/c mice. Both cells were exposed to six known phototoxic chemicals : promethazine, neutral red, chlortetracycline, amiodarone, bithionol, 8-methoxypsoralen, or non-phototoxic chemical, ALS (ammonium laureth sulfate) and then irradiated with 5 J/$cm^2$ of UVA. Cell viability ($IC_{50}$ ) was measured by neutral red uptake (NRU) assay. The ratio of $IC_{50}$ value of chemicals in the presence and absence of UVA was determined by the cut-off value. The phototoxic potential of test chemicals in NRU assay was determined by measuring the photoirriation factor (PIF) with a cut-off value of 5. In both 3T3 and HaCaT cells, all known phototoxic chemicals were positive (over 5 of PIF value), except that bithionol was found to be non-phototoxic to HaCaT cells, and ALS, non-phototoxic chemical was negative. These results suggest that Balb/c 3T3 cell was more sensitive than HaCaT cell to predict phototoxicity potential.

• Toxicological Research
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• 제7권1호
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• pp.13-20
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• 1991

To investigate the cytotoxicity and genotoxicity of the DNA alkylating agnet, mitomycin C and the antimetabolite, 5-Fluorouracil (5-FU) in cultured rat fibroblasts, the colorimetric assay of netural red (NR) for cytotoxicity and for genotoxicity, sister chromatid exchange (SCE) assay and the measurement of the rate of DNA synthesis were performed in cells cultured in media containing various concentrations of mitomycin C and 5-FU. The uptake ability of neutral red decreased does-dependently. NR90 and NR50 values of mitomycin C were 1.49 nM and 6.87mM and 5-FU were 38.4mM AND 284.4Mm respectively.

• International Journal of Oral Biology
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• 제34권3호
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• pp.143-150
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• 2009

Amino acid transporters play important roles in supplying nutrients to cells. In our current study, we investigated the expression of LAT1 and measured the amino acid uptake in ameloblast cultures to further elucidate the roles of this transporter during the differentiation of these cells. RT-PCR, observations of cell morphology, Alizaline red-S staining, and uptake analyses were performed following the experimental induction of differentiation in the cultures. LAT1 mRNA was detectable and found to gradually increase over time whereas LAT2 mRNA was not evident in the ameloblast cultures. Transcripts of 4F2hc, a cofactor of LAT1 and LAT2, were also found to be expressed in ameloblast cultures and increase with time. Amelogenin mRNA was expressed in the early stage ameloblast cultures. L-leucine uptake was observed to increase over 14 days of growth in culture. Our data suggest that LAT1 has a key role in the differentiation of ameloblasts and in providing these cells with neutral amino acids, including several essential amino acids.