• 제목/요약/키워드: Glutathione consumption assay

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Toxicity Assessment of Gas Phase in Cigarette Smoke Using Cell-free Assay

  • Park, Chul-Hoon;Sahn, Hyung-Ok;Shin, Han-Jae;Lee, Hyeong-Seok;Min, Yaung-Keun;Hyun, Hak-Chul
    • 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.

Menadione의 대사체인 Menadione-Glutathione Conjugate(MEN-SG)가 흰쥐 혈소판에 미치는 세포독성의 평가 및 MEN-SG의 안정성에 관한 연구 (Evaluation of Cytotoxicity to Rat Platelets by Menadione-Glutathione Conjugate and its Stability in Biological Assay System)

  • 서동철;정선화;이주영;김미정;정진호
    • Toxicological Research
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    • 제11권2호
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    • pp.295-302
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    • 1995
  • Menadione-ghitathione conjugate (MEN-SG), a metabolite of menadione, is known to be a redoxcycler in rat hepatocyte subcellular fraction. Therefore, it was assumed that MEN-SG could exert cytotoxlclty to ral platelets, another target tissue of menadione. We first synthesized MEN-SG, the identity of which was verified by mass, $^1{H}$-NMR and UV-visible spectra. In addition, the stability of MEN-SG was investigated in biological assay system. MEN-SG was degraded in a time-dependent manner in DMSO which had been used as a vehicle and thus, tris-HCl buffer was used as a vehicle of MEN-SG despite the low solubility in it. Perchloric acid as well as platelets itself did not affect the stability of MEN-SG. Our next attempt was the evaluation of cytotoxicity of MEN-SG in rat platelets. MEN-SG did not induce cytotoxicity to rat platelets measured by two different methods, LDH release and turbidity changes. The extents of oxygen consumption by MEN-SG in intact platelets were significantly lower than those by menadione, though it had been observed that oxygen consumptions by menadione and MENSG were similar in subcellular fractioas of platelets. These results suggest that MEN-SG is not toxic to rat platelets despite its redox cycling capacity and glutathione conjugation reaction of menadione could be regarded as a detoxification process.

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질소 화합물이 담배연기성분 및 안전성에 미치는 영향 (Effect of nitrogen compounds on the chemical composition and biological activity of mainstream smoke)

  • 신한재;박철훈;손형옥;이형석;김용하;현학철
    • 한국연초학회지
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    • 제31권2호
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    • pp.75-84
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    • 2009
  • The objective of this study was to investigate the effect of nitrogen compounds such as protein on the chemical composition and toxicity of cigarette mainstream smoke. BSA protein was treated into the tobacco leaf of original 2R4F cigarette at 1~4 % level. The studies were performed which included a bacterial mutagenicity assay and a mammalian cell cytotoxicity assay for total particulate matter(TPM), and glutathione(GSH) consumption assay for gas/vapor phase(GVP) and determination of smoke chemical constitute. Cigarettes treated with protein were observed dose-dependent increase in yield of volatiles, semi-volatiles and aromatic amines compared with control cigarette. However, carbonyl compounds such as acrolein was lower than that of control cigarette when calculated on an equal TPM basis. The cytotoxicity of TPM obtained from the protein-added cigarettes was not different from that of control cigarette. However, the mutagenicity of the TPM from protein-treated cigarettes(1~4 %) was up to 10-27 % higher than that of control. On the other hand, toxicity of GVP from protein-treated cigarette(4 %) was significantly decreased compared with control cigarette. An overall assessment of our data suggests that nitrogen compounds such as protein should be important for the chemical composition and biological activity of cigarette mainstream smoke.

Amino acid, fatty acid, and carbohydrate metabolomic profiles with ginsenoside-induced insecticidal efficacy against Ostrinia furnacalis (Guenee)

  • Liu, Shuangli;Wang, Xiaohui;Zhang, Rui;Song, Mingjie;Zhang, Nanqi;Li, Wanying;Wang, Yingping;Xu, Yonghua;Zhang, Lianxue
    • Journal of Ginseng Research
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    • 제44권4호
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    • pp.544-551
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    • 2020
  • Background: Previous studies have shown the insecticidal efficacy of ginsenosides. In the present study, we aimed to investigate the metabolic mechanism related to the inhibitory effect of panaxadiol saponins (PDSs) against the Asian corn borer Ostrinia furnacalis (Guenee). Methods: Third instar larvae of O. furnacalis were fed normal diets with different concentrations of PDSs for 4 days. The consumption index, relative growth rate, approximate digestibility, and conversion of ingested and digested food were recorded. A targeted gas chromatographye-mass spectrometry assay was performed to detect the profiles of amino acids, fatty acids, and carbohydrates in larvae of O. furnacalis. In addition, the activity of detoxification-related enzymes was determined. Results and Conclusions: PDSs decreased the consumption index, relative growth rate, approximate digestibility, and conversion of ingested and digested food in the 3rd instar larvae of O. furnacalis in a dose-dependent manner. PDSs decreased 15 free amino acids, 16 free fatty acids, and 5 carbohydrates and increased the levels of palmitoleic acid, palmitic acid, and 9-octadecenoic acid in the 3rd instar larvae. The activity of detoxification-related enzymes, such as acetylcholinesterase, glutathione S-transferase, cytochrome P450, carboxylesterase, trehalase, acid phosphatase, and alkaline phosphatase, was reduced in a dose-dependent manner in the 3rd instar larvae exposed to PDSs. These data confirmed the inhibitory effect of PDSs against growth, food utilization, and detoxification in the 3rd instar larvae of O. furnacalis and the potential for using PDSs as an efficient tool for insect pest management for O. furnacalis larvae.

Antioxidant effect of garlic and aged black garlic in animal model of type 2 diabetes mellitus

  • Lee, Young-Min;Gweon, Oh-Cheon;Seo, Yeong-Ju;Im, Ji-Eun;Kang, Min-Jung;Kim, Myo-Jeong;Kim, Jung-In
    • Nutrition Research and Practice
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    • 제3권2호
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    • pp.156-161
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    • 2009
  • Hyperglycemia in the diabetic state increases oxidative stress and antioxidant therapy can be strongly correlated with decreased risks for diabetic complications. The purpose of this study is to determine antioxidant effect of garlic and aged black garlic in animal model of type 2 diabetes. The antioxidant activity of garlic and aged black garlic was measured as the activity in scavenging free radicals by the trolox equivalent antioxidant capacity (TEAC) assay. Three week-old db/db mice were fed AIN-93G diet or diet containing 5% freeze-dried garlic or aged black garlic for 7 weeks after 1 week of adaptation. Hepatic levels of lipid peroxides and activities of antioxidant enzymes were measured. TEAC values of garlic and aged black garlic were $13.3{\pm}0.5$ and $59.2{\pm}0.8{\mu}mol/g$ wet weight, respectively. Consumption of aged black garlic significantly decreased hepatic thiobarbituric acid reactive substances (TBARS) level compared with the garlic group which showed lower TBARS level than control group (p<0.05). Activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) of garlic and aged black garlic group were significantly elevated compared to the control group. Catalase (CAT) activity of aged black garlic group was increased compared with the control group. These results show that aged black garlic exerts stronger antioxidant activity than garlic in vitro and in vivo, suggesting garlic and aged black garlic, to a greater extent, could be useful in preventing diabetic complications.

호박즙, 옥수수수염차, 팥차 및 혼합물이 식이유도 비만동물모델에서 체중과 항산화 활성에 미치는 영향 (Effect of Pumpkin, Corn Silk, Adzuki Bean, and Their Mixture on Weight Control and Antioxidant Activities in High Fat Diet-Induced Obesity Rats)

  • 박재희;이은지;박은주
    • 한국식품영양과학회지
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    • 제45권9호
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    • pp.1239-1248
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    • 2016
  • 4주간 식이로 유도된 비만 동물에서 호박즙(250 mg/kg BW), 옥수수수염차(250 mg/kg BW), 팥차(250 mg/kg BW)와 이들 혼합음료(호박즙 분말 : 옥수수수염차 분말 : 팥차 분말, 1:1:1, 250 mg/kg BW)를 9주간 투여하여 그 효능을 알아본 결과 호박즙, 옥수수수염차, 팥차 개별 섭취 시 체중감소 효과는 보였으나 그 기전은 본 연구에서 규명하지 못하였다. 그러나 혼합음료 섭취군에서는 지방조직 무게의 유의적 감소로 인한 체중 저하 효능을 확인할 수 있었다. 부고환지방조직의 표면적은 고지방식이섭취군(HFD)군에 비해 고지방식이+혼합음료섭취군(HFD+PCA)군에서 유의적으로 감소하였으며, HFD+PCA군의 아디포넥틴은 HFD군에 비해 유의적으로 증가하였다. 음료섭취군들 중 고지방식이+혼합음료섭취군(HFD+PCA)은 고지방식이섭취군(HFD)보다 ORAC value가 증가하였고, GSH-Px의 간 항산화 효소에서도 HFD군보다 HFD+PCA군이 유의적으로 증가하였다. 또한, HFD+PCA군은 고지방식이에 의한 지질과산화물 생성을 유의적으로 감소시켰으며, 비만으로 인한 산화적 스트레스에 따른 DNA 손상을 현저히 낮추었다. 따라서 산화적 스트레스에 의해 유도되는 비만 치료에 호박즙, 옥수수수염차, 팥차를 각각 섭취하기보다는 이들 세 가지를 동일한 비율로 혼합 섭취 시 각각의 항산화 효능의 상승작용에 의한 항비만 효과가 현저하게 나타낼 것으로 생각한다.