• Title/Summary/Keyword: 효소에 의한 손상

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DNA damages with Fpg/Endo Ⅲ FLARE Assay in cynomolgus monkeys exposed to stainless steel welding fume (용접흄 흡입노출 영장류에서 Fpg/Endo Ⅲ FLARE Assay를 이용한 DNA 손상 및 회복)

  • Rim, Kyung Taek;Kim, Soo Jin;Chung, Yong Hyun;Kim, Hyeon Yeong;Maeng, Seung Hee;Yu, Il Je
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.17 no.4
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    • pp.272-281
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    • 2007
  • 선박제조업을 비롯한 운송업 및 건축업 등의 다양한 분야에서 용접기술이 이용되어 옴에 따라 용접근로자들에 대한 산업보건학적 관심이 높아지고 있다. 노출정도가 다양하기는 하지만 용접흄은 6가 크롬을 비롯한 금속화합물과 유해가스, 화학물질 등을 복합적으로 포함하고 있는 스테인레스 스틸 용접흄에 대한 유전독성영향을 평가하기 위하여 흡입챔버를 이용, 실험동물인 영장류에 스테인레스 스틸 용접흄을 노출시키고 혈액 내 lymphocytes에 생성된 용접흄 노출농도 및 시간별 DNA 손상정도 및 그 회복효소를 측정함으로써, 유해성이 완전하게 확인되지 않은 용접흄에 노출되어 나타날 수 있는 암을 비롯한 심각한 건강영향을 예방하기 위한 각 지표들을 찾아 그 유용성을 비교하고자 하였다. 영장류를 노출시키기 위해 robotic arm을 장치한 영장류 흡입노출 시스템을 개발하였으며, 이 노출 시스템을 이용하여 수컷 영장류 6마리에 대해 용접흄 노출시험을 실시하였는데 실험군은 대조군 2, 저농도 ($31mg/m^3$) 노출군 2, 고농도 ($63mg/m^3$) 노출군 2마리로 구성하였고, 1일 2시간씩 일주일에 5일 동안 용접흄에 노출시켰다. 노출 농도는 지속적으로 모니터링 하였고, 노출과정 중에 영장류의 혈액을 채취하여 lymphocytes를 분리, 단세포 DNA 손상을 선별하기 위해 DNA 손상회복 효소인 E. coli formamidopyrimidine-DNA glycosylase (Fpg)와 endonuclease Ⅲ (Thymine Glycol-DNA glycosylase) 투여와 Comet asaay (single cell gel electrophoresis, 단세포겔전기영동기법)를 결합시켜 이용하는 Fpg/Endo III FLARE 분석법을 사용하였다. Fpg enzyme에 의한 olive tail moment값의 변화는 16주 노출군부터 노출부검(34주)군 까지 노출농도가 높아짐에 따른 olive tail moment 기하평균 값의 양 반응관계를 보기는 어렵지만, 고농도군의 경우 27주 노출군에서 가장 높은 olive tail moment 값을 보이고 이후 차츰 감소하였다. 한편 16주에서 22주까지의 노출기간에서는 대조군에 비해 노출군에서 DNA손상정도(olive tail moment값)는 모두 유의하게 높았으나, 6, 12, 18, 25, 31, 33, 35주간 노출하였을 때는 다른 결과를 보였다. 각 실험군의 Fpg enzyme에 의한 tail length값의 분포를 살펴볼 때, 저농도군 및 고농도군에서 27주간 노출하였을 때 가장 높은 tail length 값을 보이고 이후 차츰 감소하는 경향을 보였다. 또한 16, 22주간 노출하였을 때 대조군에 비해 노출군에서 tail length 값이 유의하게 높았으나, 20주간에서만 양 반응관계가 관찰되었고, 다른 주간에서는 양 반응 및 기간 반응관계를 나타내지는 않았다. Endo III enzyme에 의한 olive tail moment값의 변화는 기간별 노출군에서 대조군에 비해 높은 DNA손상정도(olive tail moment값)를 나타내는 결과들이 있었지만, 10, 12, 16, 22, 25, 31주간 노출하였을 때 등 상당수 노출기간에서 반응관계를 나타내지는 않았다. 각 실험군의 Endo III enzyme에 의한 tail length값의 분포를 살펴볼 때, 18, 20, 27, 33주간 노출하였을 때 대조군에 비해 노출군에서 tail length 값이 조금 높았지만, 양 반응 및 기간 반응관계를 보이지 않았고 수치의 크기가 불규칙하게 변화하였다. 즉, DNA에 있어 산화된 pyrimidine을 형성하여 손상된 부위의 염기를 제거함으로써 AP site (abasic site)를 만들고 이들이 Comet assay를 통해 break로 전환된 것을 포함한 DNA손상을 측정하기 위하여 endonuclease III (Endo III)를 첨가시킨 Endo III FLARE 분석법을 실시한 결과, 본 연구에서 나타난 결과는 용접흄 노출 영장류에서 Olive tail moment 및 tail length 공히 노출량 및 노출기간 반응관계를 볼 수 없었다. Endo III FLARE 분석법을 통한 산화적 DNA 손상지표는 영장류에 적용하기에는 적응반응현상으로 대조군과 유의한 차이도 관찰할 수 없었고 더욱이 역으로 대조군에서의 자연발생적 수치가 더 높아질 수 있어 용접흄 노출 영장류의 모니터링 지표로 사용하기에는 제한점이 있었다.

Effect of Inhibitor of Nitric Oxide Synthesis on the Ischemic Reconditioning in Isolated Heart of Rat. (NO 억제제가 허혈전처치의 심장 보호효과에 미치는 영향)

  • 유호진;조은용
    • Journal of Chest Surgery
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    • v.29 no.8
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    • pp.807-815
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    • 1996
  • The protective effect of'ischemic preconditioning'on ischemid-reperfusion injury of heart has been reported in various animal species. but without known mechAnism in detail, In An attempt to investigate the cardioprotective mechanism of ischemic preconditioning, we examined the effects of nitric oxide(UO) synthesis in preconditioned heart of rat The isolated hearts perfused by Langendorfr's method were ex- posed to 30min global ischemia followed by 30min reperfusion with oxygenated Krebs-Henseleit(K-H) sol- ution. Ischemic preconditioning was performed with three episodes of Sm n ischemia and Smin repeyfusion before the induction of prolong ischemia(30min)-reperfusion(30min). Ischemic preconditioning prevented the depression of cardiac function(left ventricular pressure .K heart rate) observed in the ischemia- reperfusion hearts and reduced the release of lactate dehydrogenase during the reperfusion period. On electromicroscopic pictures, myocardial ultrastructures wore relatively well preserved in isthemic preconditioned hearts. N6_nitro-L-arginine methyl ester(L-NAME) an inhibitor of L-arginine citric oxide pathway, was infused at a rate O.Smllmin In a dose of 10mg kg-1 before the initial ischemic preconditioning. neither the protection of cardiac function nor the reduction of LDH releAse in ischemic preconditioning hearts was altered in the presence of added L-NAME On ultrastructural finding, the preservation of morphology in ischemic preconditioning heart was not change by the pretreatment of L-UAME. The failure of the WO synthesis inhibitor to reduce t e effect of ischemic preconditioning may be related to be species specific in that NO may allot be the trigger for ischemic preconditioning in rats.

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Effects of Phenolic Compounds of Persimmon Leaves on Antioxidative System and Miscellaneous Enzyme Activities Related to Liver Function in Ethanol-Induced Hepatotoxicity of Rats (감잎의 Phenolic Compounds가 에탄올을 투여한 흰쥐의 항산화계 및 기타 효소활성에 미치는 영향)

  • 정창주;윤준식;이명렬
    • Food Science and Preservation
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    • v.11 no.1
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    • pp.79-87
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    • 2004
  • To investigate antioxidative effects of phenolic compounds separated from persimmon leaves(PL)(Diospyros kaki Thunb.) on the ethanol-induced hepatotoxicity in rat, Sprague-Dawley rats weighing 100-150 g were divided into 5 groups; control group(CON), PL(70 mg/kg) administered group(PEl), ethanol(5 mL/kg, 25%) administered group(ETH), PL(70 mg/kg) and ethanol administered group (PE2), and PL(140 mg/kg) and ethanol administered group(PE3), respectively. The antioxidative activity of persimmon leaves decreased in order of ethylacetate>interphase materials>n-butanol>chloroform>n-hexane>water fraction. The growth rate and feed efficiency ratio decreased by ethanol were gradually increased to the adjacent level of CON by administering PL. The serum activities of ALT, alkaline phosphatase and lactic acid dehydrogenase elevated by ethanol were decreased significantly. It was also observed that the activities of SOD, catalase, and GSH-Px of rat liver increased by ethanol were markedly decreased in PL administered group as compared to ETH. The GSH content of liver was decreased by ethanol, but that was increased in PE1 and PE2 compared with ETH as a dose-dependant manner. These results suggested that phenolic compounds separated from persimmon leaves have a possible protective and relievable effect on the ethanol-induced hepatotoxicity in rats.

Effect of Ramaria botrytis Methanol Extract on Antioxidant Enzyme Activities in $Benzo({\alpha})Pyrene-treated$ Mice (싸리버섯 메탄올 추출물이 벤조피렌을 투여한 마우스의 항산화 효소 활성에 미치는 영향)

  • Kim, Hyun-Jeong;Lee, Kap-Rang
    • Korean Journal of Food Science and Technology
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    • v.35 no.2
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    • pp.286-290
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    • 2003
  • Effects of Ramaria botrytis methanol extract on hepatotoxicity in $benzo({\alpha})pyrene(B({\alpha})P)-treated$ mice were investigated. R. botrytis methanol extract was intraperitioneally injected once a day for successive 5 days, followed by treatment with $B({\alpha})P$ on the fifth day. Antioxidant activities of R. botrytis methanol extract were examined by measuring the free radical-scavenging effect on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical. In DPPH method, R. botrytis methanol extract showed strong antioxidative activies. The increased activities of superoxide dismutase, catalase, and glutathione peroxidase after $B({\alpha})P-treatment$ were decreased by treatment of R. botrytis methanol extract. Glutathione content and glutathione S-transferase activity depleted by $B({\alpha})P$ were significantly increased, but elevation of lipid peroxide content induced by $B({\alpha})P$ was decreased by R. botrytis methanol extract. These results suggest that R. botrytis methanol extract is believe to be a possible protective effect against $B(\alpha)P-induced$ hepatotoxicity in mice.

Effect of Larva Extract of Allomyrina dichotoma on Carbon Tetrachloride-induced Hepatotoxicity in Mice (장수풍뎅이 유충 추출물이 사염화탄소에 의한 마우스의 간 손상에 미치는 영향)

  • Choi, Yong-Hwa;Lee, Ki-Yeol;Yang, Kyung-Mi;Jeong, Yun-Mi;Seo, Jung-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.10
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    • pp.1349-1355
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    • 2006
  • This study was conducted to investigate the effect of methanol extract of Allomyrina dichotoma larva (MEAL) on carbon tetrachloride $(CCl_4)-induced$ hepatotoxicity in mice. ICR mice were divided into 5 groups [Vehicle control, $CCl_4\;(10{\mu}g/g)$ alone, $CCl_4$ plus a low dose $(50{\mu}g/g)$ of MEAL, $CCl_4$ plus a high dose $(100{\mu}g/g)$ of MEAL]. Silymarin $(2{\mu}g/g)$ was used as the reference in the experiment. Administration of MEAL tended to decrease the serum alanine transaminase (ALT) activity induced by $CCl_4$ treatment in mice. Hepatic concentration of thiobarbituric acid-reactive substances (TBARS) in a high-dose group of diet decreased to the level of silymarin-treated group. Hepatic activity of glutathione S-transferase in MEAL-treated group was lower than that of $CCl_4-treated$ group. Serum concentration of bilirubin was significantly increased by $CCl_4$ treatment, but MEAL or silymarin recovered the level. These results suggest that MEAL may exert the protective effect against $CCl_4-induced$ hepatotoxicity in mice. However, more intensive studies would be needed to elucidate the protective mechanism of the beetle on hepatotoxicity of mice.

Effects of Godulbaegi Leaf Extracts on $CCl_4$-Induced Hepatotoxicity in Rats (고들빼기 잎추출물이 흰쥐의 사염화탄소에 의한 간손상에 미치는 영향)

  • 배송자;김남홍;하배진;정복미;노승배
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.1
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    • pp.137-143
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    • 1997
  • This study was carried out to investigate the effect of godulbaegi leaf extracts on detoxication of liver injury in $CCl_4$-treated rats. We divided normal group(5) and $CCl_4$-treated group and then $CCl_4$- treated group was divided into 6 groups; only $CCl_4$-treated group(CS), aqueous extract pretreated group (CSA), n-butanol extract pretreated group(CSB), Tween 80 pretreated group(CT), n-hexane extract pretreated group(CTH) and ethyl acetate extract pretreated group(CTE). The ratio of liver weight per body weight and the activity of GPT in hexane extract group(CTH) were lowest, similar to the results of S. Godulbaegi n-hexane extract(IS) groups intoxicated by $CCl_4$ had lower values of MDA than CS and CT which are control groups. Histological finding of liver tissue revealed less of necrosis in IS extracts groups than in control groups(CS, CT). From these results, IS extracts could predominently prevent hepat-otoxicity of rats. Especially, hexane extract was effective on the detoxication of liver injury among the other extracts.

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A Study on the Effect of Injection Frequency on the Liver Damage in Rats (Bromobenzene의 투여 횟수에 따른 간독성의 차이)

  • 이상희;전태원;윤종국
    • Biomedical Science Letters
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    • v.6 no.1
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    • pp.29-36
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    • 2000
  • To investigate the effect of injection frequency of bromobenzene on the liver damage, bromobenzene (400 mg/kg, i.p.) was given daily to rats for six days. All experimental animals were sacrificed at 24 hours after the last injection. Morphological changes of the liver were observed under a light microscopic examination. Functional changes of the liver were evaluated by the measurement of alanine aminotransferase activity. To clarify the cause of discrepancy in liver damage, hepatic glutathione (GSH) content, glutathione S-transferase (GST) and aniline hydroxylase (AH) activities were determined. In the experiments of daily bromobenzene treatments, the sacrificed animals at six day (6 time-injected animals) showed slighter liver damage than those sacrificed at 3 day (3 time-injected ones), based on the liver morphological or functional findings; the decreasing ratio of GSH content and increasing ratio of liver GST and AH activities in the 6 time-injected group were higher than those in the 3 time-injected one.

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The Protective Effects of Cornus walteri Wanger Leaves against UV Induced Cellular Damage in Human Fibroblast (자외선에 의한 세포손상에 대한 말채나무잎의 보호효과)

  • Park, Hyun-Chul;Jung, Taek Kyu;Yoon, Kyung-Sup
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.1
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    • pp.1-7
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    • 2015
  • Cornus walteri Wanger has been used in folk medicine in Korea. Ultraviolet (UV) irradiation has been known as a major cause of photo damage in skin. In the present study, research on how to cure damaged cells by UVB was conducted using an extract of Cornus walteri Wanger leaves (CWE), which was treated with an enzyme. CWE was applied to human dermal fibroblasts (HDFs) affected by UVB. UVB-irradiated HS68 cells showed increased caspase-3 activity, phosphorylation of p53, ${\gamma}H2AX$, cyclobutane pyrimidine dimers (CPDs) formation, and DNA fragmentation compared with non-irradiated cells. However, all these effects were inhibited by treatment with CWE for 12 h after UVB irradiation. Furthermore, CWE has proved not to cause primary skin irritation through the human patch test. Collectively, these results suggest that CWE could be a new potential candidate as photoprotective agent against UVB-induced cellular damage in HDFs.

Effect of the Ethanol Extract of Cassia tora L. on Antioxidative Compounds and Lipid metabolism in Hepatotoxicity of Rats-induced by Ethanol (결명자 에탄올추출물이 알코올 투여 흰쥐의 항산화물질 및 지질대사에 미치는 영향)

  • 최현숙;차선숙;나명순;신길만;이명렬
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.6
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    • pp.1177-1183
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    • 2001
  • This study was done to investigate the effects of ethanol extract of Cassia semen (Cassia tora L.) on the activities of hepatic oxygen free radicals metabolizing enzymes and blood lipid profile in rats of hepatotoxicity induced by ethanol. Sprague-Dawley male rats weighing 100~160 g were divides into 5 groups; control grouts (CON), Cassia semen ethanol extracts (200 mg/kg) treated group (CEL), ethanol (10 mL/kg, 35%) treated group (ETH), Cassia semen ethanol extracts (200 mg/kg) and ethanol treated group (CE1 ) and Cassia semen ethanol extracts (400 mg/kg ) and ethanol treated group (CE2), respectively. Compared with ETH, the growth rate of CE1 and CE2 were to be increased tendency, and in blood levels of total cholesterol, LDL-cholesterol and the activities of alanine aminotranferase and asparate aminotranferase elevated by ethanol were significantly decreased (p<0.05). It was observed that the activities of superoxide dismutase, catalase, xanthine oxidase and glutathione peroxidase of rat liver increased by ethanol, were more decreased by the treatment of Cassia semen ethanol extract than the only ethanol-treated group. The content of glutathione depleted by ethanol treatment was increased in CE1 and CE2. TBA-reactants of liver increased by ethanol were decreased in CE1 and CE2, compared with ethanol-treated group. These results suggested that ethanol extract of Cassia semen may influence upon the ability of oxygen free radical detoxication and lowering of blood lipid level on ethanol-induced hepatotoxicity in rat.

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Immunostimulatory effects of enzymatic porcine placental hydrolyzate against cyclophosphamide-induced immunosuppressed model (돈태반 효소 가수분해물의 cyclophosphamide에 의한 면역 저하 동물 모델에 미치는 면역 증진 효과)

  • Kim, Keun Nam;Kim, Min Ju;Yoon, Sun Myung;Kwon, Min Joo;Shin, Dong Yeop;Lee, Hak Yong;Park, Young Mi
    • Korean Journal of Food Science and Technology
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    • v.54 no.2
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    • pp.155-162
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    • 2022
  • This study investigated the immunostimulatory effect of enzymatic porcine placental hydrolyzate (EPPH) in cyclophosphamide (Cy)-treated rats. This effect of EPPH prevented Cy-induced decreases in body, spleen, and thymus weights and natural killer (NK) cell activity. The numbers of immune cells, such as white blood cells, granulocytes, and lymphocytes, and mid-range absolute counts were significantly higher compared to the control group. The levels of tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1β, IL-2, IL-12, and immunoglobulin G (IgG) were notably reduced by Cy, while EPPH prevented these effects. Histopathological analysis of spleen samples revealed the protective effect of EPPH against Cy-induced immunosuppression. The findings demonstrate that EPPH can alleviate immunosuppression by cell viability, tissue damage, and regulation of the levels of cytokines. EPPH may have value as a component of immunostimulatory agents or an ingredient in functional foods.