• 한국식품위생안전성학회지
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• 제14권3호
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• pp.259-264
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• 1999

Xylooligosaccharide의 세균에 대한 돌연변이 유발성을 검색하기 위하여 Salmonella typhimurium의 히스티딘 요구성균주 TA100, TA1535, TA98 및 TA1537의 4개 균주와 대장균 Escherichia coli의 트립토판 요구성 균주인 WP2 uvrA를 이용해 복귀돌연변이 시험을 실시하였다. 시험 물질은 증류수에 용해하여 처리하였으며, 대사 활성계 미적용 및 적용하에 $5000\;\mu\textrm{g}/plate$를 최고농도로 하고 공비 2로서 5단계 농도군(313, 625, 1250, 2500 및 $5000\;\mu\textrm{g}/plate$)과 음성 및 양성 대조군으로 시험군을 구성해 본 시험을 실시하였다. 시험 결과 시험물질을 처리한 모든 농도군에서 복귀돌연변이 집락 수는 음성대조군과 비슷한 정도로 관찰되었다. 이상의 결과에서, xylooligosaccharide는 본 시험 조건 하에 사용한 균주들에 복귀돌연변이를 유발하지 않는 것으로 사료되었다.

• Applied Biological Chemistry
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• 제37권3호
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• pp.203-209
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• 1994

$^{14}C-{\alpha}-endosulfan$(100 nM, $41\;{\mu}g$)을 공시어의 각 조직(간, 신장, 소화관) 추출액에 처리하여 대사물질의 생성과 이에 미치는 cofactor의 영향을 시험관내 반응으로 비교한 결과, 105,000 g soluble fraction에서 phase I system에 해당되는 cofactor의 첨가시 공시어의 간 조직에서 가장 높은 대사물질 생성율을 보였고 phase II system에서는 NADPH를 cofactor로 첨가하였을 때 간과 신장조직에서 높은 대사물질 생성율을 보였다. 하지만 소화관 조직에서는 GSH 단독처리나 GSH+NADPH의 혼합처리시 높은 대사물질 생성율을 보였다. 또한 microsomal fraction에서는 phase I system에 해당되는 cofactor의 첨가시 공시어의 소화관 조직에서, phase II system의 경우 NADPH 단독처리나 NADPH+GSH 혼합처리시 대사물질의 생성이 가장 많았다. 이러한 결과로 보아 공시어의 간과 신장에서는 MFO에 의한 대사작용이, 소화관에서는 GST에 의한 대사작용이 활발하다고 할 수 있었다. 공시어의 각 조직에서 높은 생성율을 보인 대사물질은 EA(endosulfan alcohol), EE(endosulfan ether), EHE(endosulfan hydroxyether)이었으며 in vivo 시험에서와 마찬가지로 endosulfan sulfate는 검출되지 않았다. 이 실험에서 ${\alpha}-endosulfan$은 잉어체내에서 ${\beta}-endosulfan$으로 이성화되었다가 EA로 가수분해되거나 직접 EA로 가수분해되어 다시 EE나 EHE로 산화되며 EHE는 EL(endosulfan lactone)로 산화되는 것으로 제안할 수 있었다.

• 대한약리학회지
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• 제16권2호
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• pp.45-53
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• 1980

Lipid peroxidation in vitro has been identified as a basic deteriorative reaction in cellular mechanism of aging processes, such as air pollution oxidant damage to cell and to the lung, chlorinated hydrocarbon hepatotoxicity. Many experimental evidences were reported by several investigators that lipid peroxidation could be one of the principle causes for the hepatotoxicity produced by $CCl_4$. It is now reasonably established that $CCl_4$ is activated to a free radical in vivo, that lipid peroxidation occurs very quickly in microsomes prepared from damaged livers, that the peroxidation is associated with loss of enzyme activity of microsomes, and that various antioxidants can protect animals against the hepatotoxic effect of $CCl_4$. Recent studies have drawn attention to some other feature of microsomal lipid peroxidation. Incubation of liver microsomes in the presence of NADPH has led to a loss of cytochrome $P_{450}$. However, the presence of an antioxidant prevented lipid peroxidation and preserved cytochrome $P_{450}$. Decrease of cytochrome $P_{450}$ in microsomes under in vitro incubation can be enhanced by $CCl_4 and these changes were parallel to a loss of microsomal polyunsaturated fatty acid and formation of malonaldehyde. The primary purpose of this experiment was to study the effect of riboflavin tetrabutylate on lipid peroxidation, specially, the relationship between lipid peroxidation and drug metabolizing enzyme system which is located in smooth endoplasmic recticulum as well as the effect of ritoflavin tetrabutylate on drug metabolizing enzyme system of animal treated with $CCl_4$. Albino rats were used for experimental animal. In order to induce drug metabolizing enzyme system, phenobarbital was injected intraperitoneally. $CCl_$ and riboflavin tetrabutylate were given intraperitoneally as solution in olive oil. Microsomal fraction was isolated from liver of animals and TBA value as well as the activity of drug metabolizing enzyme were measured in the microsomal fractions. The results are summerized as following. 1) The secobarbital induced sleeping time of $CCl_4$ treated rat was about 2 times longer than that of the control group. However, the pretreatment with riboflavin tetrabutylate inhibited completely the lengthened sleeping time due to $CCl_4$ treatment. Furthermore TBA value was significantly increased in $CCl_4$ treated rat in comparison to control group tut the increase of TBA value was prevented by the pretreatment with riboflavin tetrabutylate. On the other hand, the activity of hepatic drug metabolizing enzyme was decreased in $CCl_4$ group, however, the pretreatment with riboflavin tetrabutylate also prevented the decrease of the enzyme activity caused by $CCl_4$. 2) The effect of riboflavin tetrabutylate on TBA value and the activity of drug metabolizing enzyme in vitro was similar to in vivo results. Incubation of liver microsome from rat in the presence of $CCl_4$, $Fe^{++}$, or ascorbic acid has led to the marked increase of TBA value, however, the addition of riboflavin tetrabutylate in incubation mixture prevented significantly the increase of TBA value, suggesting the inhibition of lipid peroxidation. In accordance with TBA value, the activity of drug metabolizing enzyme was inhibited in the presence of $CCl_4$, $Fe^{++}$, ascorbic acid but the addition of riboflavin tetrabutylate protected the loss of the enzyme activity in microsome under in vitro incubation.

• 한국식품영양학회지
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• 제11권1호
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• pp.114-122
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• 1998

표고버섯의 열수 가용성 다당류(PS)와 간 장해물질이며 발암성을 갖는 butter yellow를 흰쥐의 식이에 6주간 투여하여 혈장과 간의 효소활성 및 과산화지질의 함량을 측정한 결과는 다음과 같다. 1. 혈장 중 GOT 활성은 better yellow 첨가군(BO군)보다 PS의 투여군(BP군 : BO+PS)이 비교적 낮은 수준을 나타내었으며 ${\gamma}$-GTP와 GPT 활성에서도 BO군에 비하여 BP군에서 유의하게 낮은 활성을 나타냈다. 2. 간장 중 glutathion peroxidase, catalase 및 lactate dephdregenase 활성 또한 기초식이군(NO군)에 PS를 토여한 군(NP군)은 NO군과 유의한 차이가 없었으나 BP군은 BO군에 비하여 유의하게 낮은 수준이었다. SOD활성은 각 군간에 유사한 수치로 유의적인 차리를 보이지 않았다. 3. 간장의 microsomal 분획분 및 cytosol 분획분 중의 glutathione S-transferase 활성도는 BP군이 BO군보다 유의하게 낮은 수준을 보였으나 better yellow를 첨가하지 않은 NO군에 비하여 상당히 높은 수준을 나타내었다. 4. 간장 내 glutathione과 malondialdehyde의 함량 또한 PS의 투여로 생성이 저하되는 것으로 나타났다. 이상의 결과를 보아 표고버섯의 열수 추출물은 생체조직의 지질의 과산화를 방지하므로써 better yellow에 의한 간장장애의 치료 또는 독성을 완화하는 효과가 있는 것으로 사료된다.

• Applied Biological Chemistry
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• 제41권1호
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• pp.47-52
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• 1998

3-Hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) 환원효소는 식물의 병원균 방어물질 (phytoalexin), 광합성 색소 (the phytol of chlorophyll), 성장 호르몬 (abscisic acid와 gibberellin 등) 및 스테롤 (phytosterol) 등의 생합성에 관여하는 주효소이다. 암조건 하에서 발아 후 4일째의 벼 유묘 microsome을 재료로 비이온성 detergent 인 Brij W-1 (final 0.4%)을 사용하여 가용화 시킨 후, DEAE-Sephadex A-50 크로마토그래피 칼럼과 hexaneagarose를 matrix로 기질인 HMG-CoA를 결합시켜 제조한 친화성 크로마토그래피 칼럼을 이용하여 세포막 결합효소인 HMG-CoA 환원효소를 정제하였다. 정제된 HMG-CoA 환원효소의 최종 회수율은 7.14% 였고, 분자량은 10% SDS-PAGE에서 55 kDa이였다 HMG-CoA 환원효소의 최적 반응 온도는 $37^{\circ}C$, 최적 반응 pH는 6.9였고, 기질인 HMG- CoA에 대한 HMG-CoA 환원효소의 $K_m$ 및 $V_{max}$ 값은 $180\;{\mu}M$과 107 pmol/mg, 수소공여체인 NADPH에 대한 $K_m$과 $V_{max}$값은 $810\;{\mu}M$과 32.1 pmol/min/mg이였다.

• Applied Biological Chemistry
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• 제40권2호
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• pp.112-116
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• 1997

소의 뇌에서 neurosteroid의 지방산 에스터를 합성하는 neurosteroid acyltransferase(NSAT)의 뇌에서의 부위별, 세포분획별 분포 및 효소적 특성을 연구하였다. 소의 뇌 NSAT 활성도는 소뇌(小腦)에서 가장 높았으며 그 다음이 중뇌(中腦)이었다. 가장 활성도가 낮은 부위는 대뇌(大腦)로 소뇌(小腦)의 50% 수준이었다. 또한 소뇌 (小腦) microsome에서의 활성도가 다른 세포분획에 비해 매우 높아 NSAT가 microsomal 효소임을 알 수 있었다. NSAT의 최적온도는 $40^{\circ}C$이었으며, pH=4.9에서 최적 pH를 나타냈다. DHEA를 기질로 하였을 때 NSAT의 $K_m$과 $V_{max}$는 각각 $32.6\;{\mu}M$과 4.86 nmole/mg protein/h였다. 또한 DHEA를 기질로 하였을 때 pregnenolone $({\Delta}5P)$은 NSAT에 대하여 경쟁적 저해제로, testosterone은 비경쟁적 저해제로 작용하였다. ${\Delta}5P$와 testosterone의 Ki는 각각 $22.8\;{\mu}M$과 $28.2\;{\mu}M$이었다. 따라서 소의 NSAT는 C-3의 ${\beta}-hydroxyl$ 기와 C-17의 ${\beta}-hydroxyl$ 기를 에스터화하는 과정에서 다른 conformation을 가짐을 시사한다.

• 한국식품위생안전성학회지
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• 제20권3호
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• pp.175-178
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• 2005

Clean Watural은 주성분이 참나무로부터 추출정제한 목초액과 프로폴리스로 구성된 새로운 천연 살균소독제이다. Salmonella typhimurium TA98, TA100, TA102, TA1535 및 TA1537 등의 5종의 균주를 이용한 복귀돌연변이성 시험을 수행한 결과, 모든 균주에서 대조군과 비교하여 시험용량군 사이에 돌연변이 유발성이 관찰되지 않았다. 그러나, 앙성대조물질에서는 대조군과 비교하여 통계학적으로 유의한 균주의 증가가 관찰되었다. 따라서, 시험물질인 천연 살균소독제, Clean Natulat은 돌연변이 유발 가능성이 없는 것으로 평가되었다.

• 한국연초학회지
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• 제13권1호
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• pp.19-26
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• 1991

Ames test using special strains which are histidine requiring mutant of Salmonella typhimufium , is widely utilized as short-term bioassay to evaluate the genotoxicity of chemicals. This method requires the liver microsome(5-9 fraction) to provide mammalian metabolism of the compounds, Therefore, the mutagenic potency of the chemicals is affected by not only the intrinsic properties of them but also the efficiency of the in vitro microsomal activation system. For this reason, the complex mixtures such as environmental pollutants from occupational sources, natural products or cigarette smoke condensates(CSC) , might be often appeared the false results. Induction of sister chromatid exchange(SCE) in cultured cells is known as another sensitive and powerful tool for the measurement of genotoxicity and the method has also an advantage which is able to apply to the in vitro and in vivo systems. In the present study, the inducibilities of revertant colonies in tester strain TA98 and SCE in human Iymphocytes by positive controls and total particulated materials(TPM) obtained from various brand(domastic and imported) cigarettes were compared in order to investigate whether the results in Ames test are in agreement with those in SCE analysis. The mutagenic activities of well known mutagens such as B(a)P showed excellent dose-response in the both methods although the induction mechanism was different each other, but cyclophosphamide resulted such effect only in SCE analysis. Most TPM tested showed a similar pattern in the mutagenic activities in those methods. However, only two(one imported brand and one domestic sample cigarettes) among the TMP obtained from various cigarettes appeared the higher induction in SCE than Ames test.

• The Korean Journal of Physiology and Pharmacology
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• 제2권5호
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• pp.601-609
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• 1998

The present study was undertaken to examine the role of phospholipase $A_2\;(PLA_2)$ in oxidant-induced inhibition of phosphate transport in primary cultured rabbit renal proximal tubule cells. Uptakes of phosphate and glucose were dose-dependently inhibited by an oxidant t-butylhydroperoxide (tBHP), and the significant inhibition appeared at 0.025 mM of tBHP, whereas tBHP-induced alterations in lipid peroxidation and cell viability were seen at 0.5 mM. tBHP stimulated arachidonic acid (AA) release in a dose-dependent fashion. A $PLA_2$ inhibitor mepacrine prevented tBHP-induced AA release, but it did not alter the inhibition of phosphate uptake and the decrease in cell viability induced by tBHP. tBHP-induced inhibition of phosphate transport was not affected by a PKC inhibitor, staurosporine. tBHP at 0.1 mM did not produce the inhibition of $Na^+-K^+-ATPase$ activity in microsomal fraction, although it significantly inhibited at 1.0 mM. These results suggest that tBHP can inhibit phosphate uptake through a mechanism independent of $PLA_2$ activation, irreversible cell injury, and lipid peroxidation in primary cultured rabbit renal proximal tubular cells.

• Journal of Acupuncture Research
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• 제19권5호
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• pp.199-208
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• 2002

Objective : This study was undertaken to determine if Carthami Semen Aquacupunc- ture(CSA) exerts protective effect against alterations in membrane transport function rabbits with mercury chloride(HG)-induced acute renal failure. Methods : The administration of Hg at a subcutaneous single dose of 10 mg/kg caused a reduction in GFR and an increase in fractional Na excretion, indicating generation of acute renal failure. When CSA were given for 7 days prior to Hg administration, such changes were significantly attenuated. The fractional excretion of glucose and phosphate was increased in rabbits treated with Hg alone. Results : The increase in rabbits treated with Hg following CSA are significantly lower than that in animals treated with Hg alone. Uptakes of glucose and phosphate in purified isolated brush-border membrane and Na-K-ATPase activity in microsomal fraction were inhibited in rabbits treated with Hg alone. Such changes were prevented by CSA. Uptakes of organic ions, PAH and TEA, in renal cortical slices were inhibited by the administration of Hg, which was prevented by CSA. Exposure of renal cortical slices to Hg in vitro caused an increased LDH release and lipid peroxidation, which was significantly prevented by CSA extract. Conclusions : These results indicate that the administration of Hg causes impairment in reabsorption of solutes in the proximal tubule via the generation of reactive oxygen species. CSA provides the protection against the impairment in proximal reabsorption, and its effect may be resulted from its antioxidant effect.