• Title/Summary/Keyword: 효소에 의한 손상

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Effect of Salviae Radix on Cell Death and DNA Damage in Renal Proximal Tubular Cells Exposed to $H_2O_2$ ($H_2O_2$에 노출된 신장 근위 세뇨관 세포에서의 세포 사망 및 DNA 손상에 대한 단삼의 효과)

  • Sung-Dae, Kim;Ji-Cheon, Jeong
    • The Journal of Korean Medicine
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    • v.22 no.3
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    • pp.21-30
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    • 2001
  • 목적 : 이전 연구에서 단삼 추출액이 강력한 항산화 작용이 있음을 확인한 바 있어 단삼 추출액이 신장세뇨관 세포에서 oxidant에 의한 세포사망 및 DNA 손상을 방지하는 지를 조사하고 이러한 효과가 지질의 과산화를 억제하는 효과에 기인하는 지를 시험하였다. 방법 : 신장 근위세뇨관 세포 유래 세포주인 opossum kidney (OK)세포를 이용하여 세포 사망은 frypan blue exclusion방법을 이용하여 평가하였고, DNA손상 정도는 double stranded DNA의 파괴를 측정하여 평가하였다. Oxidant 약물 모델로는 $H_2O_2$를 사용하였다. 결과 : $H_2O_2$는 적용시 간과 농도에 비례하여 세포 사망을 유도하였다. 단삼 추출액은 0.05% 농도에서 $H_2O_2$에 의한 세포사망 및 DNA 손상을 방지하였다. 이러한 방지효과는 $H_2O_2$ 제거 효소인 catalase와 철 착염제인 deferoxamine에 의해서도 나타났다. 그러나 강력한 항산화제인 DPPD는 $H_2O_2$에 의한 세포 사망이나 DNA손상을 방지하지 못하였다. $H_2O_2$는 세포내 ATP 농도를 감소시켰으며. 이러한 감소는 poly (ADP-ribose) polymerase억제제인 3-aminobenzamide에 의해 방지되었으나 단삼 추출액에 의해서는 영향을 받지 않았다. 3-aminobenzamide는 $H_2O_2$에 의한 세포 사망을 방지하였다. $H_2O_2$는 지질의 과산화를 증가시켰으며, 이러한 변화는 단삼 추출액과 DPPD에 의해 방지되었다 결론 : OK 세포에서 $H_2O_2$에 의한 세포사망과 DNA 손상에는 지질의 과산화가 중요한 역할을 하지 않으며, 단삼 추출액의 $H_2O_2$에 의한 세포 사망과 DNA 손상 방지 효과는 항산화 작용이 아닌 다른 기전에 기인하는 것으로 사료된다.

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Expression of Matrix Metalloproteinase-2 and Tissue Inhibitor of Metalloproteinase-2 in Radiation Exposed Small Intestinal Mucosa of the Rat (방사선조사를 받은 흰쥐 소장 점막의 손상과 재생과정 중 금속단백효소 및 억제자의 발현)

  • Kwag, Hyon-Joo;Lee, Kyoung-Ja;Rhee, Chung-Sik
    • Radiation Oncology Journal
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    • v.21 no.1
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    • pp.66-74
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    • 2003
  • Purpose : The matrix metalloprotelnases (MMPs) are a family of enzymes whose main function is the degradation of the extracellular matrix. Several studies have revealed that MMPs and TIMPS are related to the wound heating process and in photoaging caused by ultraviolet Irradiation. However, the expressions of MMP and TIMP after irradiation have not, to the best of our knowledge, been studied. This study investigates the expressions of MMP-2 and TIMP-2 in rat Intestinal mucosa following irradiation. Materials and Methods : The entire abdomen of Sprague-Dawley rats was irradiated using a single dose method. The rats were sacrificed on day 1, 2, 3, 5, 7 and 14 following irradiation. Histopathological observations were made using hematoxilin & eosin staining. The expressions of MMP-2 and TIMP-2 were examined using immunohistochemistry, Irnrnunoblotting and ELISA. Results : Radiation induced damage associated with atrophic villi, and infiltration of inflammatory cell was observed from the first postirradiation day, and severe tissue damage was observed on the second and the third postirradiation days. An increase in mitosis and the number of regenerating crypts, as evidence of regeneration, were most noticeable on the fifth postirradiation day. From the immunohistochemlstry, the MMP-2 expression was observed from the first postirradiation day, but was most conspicuous on the third and the fifth postirradiation days. The TIMP-2 expression was most conspicuous on the fifth postirradiation day. From the irnrnunoblotting, the MMP-2 expression was strongly positive on the third postirradlatlon day, and that of TIMP-2 showed a strong positive response on the fifth postirradiation day. In ELISA tests, the expressions of MMP-2 and TIMP-2 were increased in the postirradiation groups compared to those of the normal controls, and showed a maximum increase on the fifth postirradiatlon day. These results were statistically significant. Conclusion : The expressions of MMP-2 and TIMP-2 were increased in the intestinal mucosa of the rats following irradiation, and these results correlated with the histopathological findings, such as tissue damage and regeneration. Therefore, this study suggests that MMP-2 and TIMP-2 play roles in the mechanisms of radiation-induced damage and regeneration of intestinal mucosa of rats.

Isolation and Physicochemical Properties of Rice Starch from Rice Flour using Protease (단백질분해효소에 의한 쌀가루로부터 쌀전분의 분리 및 물리화학적 특성)

  • Kim, ReeJae;Oh, Jiwon;Kim, Hyun-Seok
    • Food Engineering Progress
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    • v.23 no.3
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    • pp.193-199
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    • 2019
  • This study aimed to investigate the impact of protease treatments on the yield of rice starch (RST) from frozen rice flours, and to compare the physicochemical properties of RST by alkaline steeping (control) and enzymatic isolation (E-RST) methods. Although the yield of E-RST, prepared according to conditions designed by the modified 23 complete factorial design, was lower than the control, the opposite trends were observed in its purity. E-RST (RST1, isolated for 8 h at 15℃ with 0.5% protease; RST2, isolated for 24 h at 15℃ with 1.5% protease; RST3, isolated for 24 h at 15℃ with 0.5% protease) with the yields above 50% were selected. Amylose contents did not significantly differ for the control and RST2. Relative to the control, solubilities were higher for all E-RST, but swelling power did not significantly differ for E-RST except for RST1. Although all E-RST revealed higher gelatinization temperatures than the control, the reversed trends were found in the gelatinization enthalpy. The pasting viscosities of all E-RST were lower than those of the control. Consequently, the enzymatic isolation method using protease would be a more time-saving and eco-friendly way of preparing RST than the alkaline steeping method, even though its characteristics are different.

Activation of Cytosolic Phospholipase $A_2$ by Methyl Mercury($CH_3$HgCl) in Madin Darby Canine Kidney (MDCK) cells

  • Kang, Mi-sun;Seo, Ji-Heui;Huh, Don-Hang;Kim, Dae-Kyong
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1997.04a
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    • pp.79-79
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    • 1997
  • 자연계에 존재하는 수은중 유기수은은 생태계 먹이사슬을 통하여 체내의 여러장기에 축적되어 조직손상을 일으키는 것으로 잘 알려져 있다. 그러나 이러한 세포독성에 대한 정확한 생화학적 기전에 대해서는 자세히 알려진 바가 없다. 포스포리파아제 $A_2$(PLA$_2$)는 세포막의 인지질로부터 Arachidonic acid (AA)와 Lysophospholipid를 유리시키는 효소로 최근 세포손상과 관련하여 그 역할이 주목되고 있으며, 극히 최근, 일차배양 소뇌신경세포를 이용한 연구에서 메칠수은처리에 의해 세포독성의 지표인 Lactate dehydrogenase (LDH)의 유리와 함께 AA 유리가 증가되는 것이 관찰되었으나 여러형태의 PLA$_2$중 어느형태의 효소가 관련되어 있는지, 또한, 그 자세한 기전에 대해서는 불분명한 점이 많다. 본 연구에서는 신장세포의 일종인 MDCK세포를 이용하여 메칠수은의 처리에 의한 PLA$_2$의 활성화 및 그 생화학적인 기전을 구명하고자 하였다. [$^3$H]AA를 MDCK세포의 배양액에 첨가하여 라벨링한 후 메칠수은을 처리하였을때 [$^3$H]AA가 대조군에 비해 농도의존적 및 경시적으로 현저하게 증가하였으며 동시에 LDH의 유리도 함께 관찰되었다. 이러한 [$^3$H]AA의 유리 증가는 세포질 PLA$_2$에 특이적인 저해제로 알려진 AACOCF$_3$의 전처리에 의해 거의 완전히 억제되었으나 LDH의 유리는 오히려 증가하였다. 또한, 글루타치온(GSH)의 전구체인 NAC (N-Acetyl Cysteine)에 의해 [$^3$H]AA의 유리는 부분적으로 감소하였으나, LDH의 유리는 변함이 없었다. 돼지비장이나 MDCK 세포에서 얻어진 세포질 PLA$_2$에 메칠수은을 직접 처리하였을때는 오히려 PLA$_2$의 활성은 감소되었다. 위의 결과들로부터 메칠수은에 의한 [$^3$H]AA의 유리 증가는 세포질 PLA$_2$효소에 대한 직접적인 작용이 아니라 세포내 -SH기의 차단이나 Oxidative Stress에 의해 간접적으로 활성화되는 것으로 예상되며, 세포질 PLA$_2$에 의해 유리된 AA의 세포독설과 관련된 세포내의 역할에 대해 의문이 제기되었다.

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An Influence of Protease on Damage of Fiber (Protease가 섬유의 손상에 미치는 영향)

  • Song, Gyeong-Heon;Yang, Jin-Suk;Choe, Jong-Myeong
    • Journal of the Korean Society of Clothing and Textiles
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    • v.22 no.2
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    • pp.224-232
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    • 1998
  • Protease is mixtured in detergent to remove protein-soil easily. It must not act on the any fiber except protein-soil during laundry. So the purpose of this study is to investigate how protease is affect the fiber, particulary the protein-fiber. For this purpose, silk, wool and nylon are selected as samples, and the extent of the damage was estimated as tensile strength and surface condition (that is fibrillation). The results are as follows. The tensile strength of fiber treated with protease were lowered at enzyme concentration 0.1%, temperature 4$0^{\circ}C$ , and, as washing time was longer, it was lowered more. And it was showed that the surface of fibers were fiblliated by protease during washing. From this results, it was found that protease damaged protein-fiber. The damage of silk was the largest of all, and wool was less damaged than silk, because it has the scale (cuticle) on the outside. Additionary, an influence of surfactant on damage of fiber was little about three fibers, but, the fibers were damaged more by the binary nonionic-surfactant and protease mixture than by protease only.

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Effects of Enzyme Treatment in Steeping Process on Physicochemical Properties of Wet-Milled Rice Flour (효소 전처리에 의한 습식제분 쌀가루의 이화학적 특성)

  • Kim, Rae-Young;Park, Jae-Hee;Kim, Chang-Soon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.9
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    • pp.1300-1306
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    • 2011
  • This study investigated the physicochemical characteristics of wet-milled rice flour treated with pectinase and cellulase in a steeping process. Enzyme treatments were used as follows: pectinase 0.05%, cellulase 0.05%, and mixed enzyme treatments 0.05~0.2%. For particle distribution, rice flour E-treated with mixed enzymes (pectinase 0.05% and cellulase 0.05%) was the finest at 48.3% particle distribution less than $53\;{\mu}m$. Protein contents and damaged starch were reduced by enzyme treatments. Damaged starch was the lowest (12.1%) in rice flour E compared with non-enzyme treatment (18.1%). Amylose content, water binding capacity, solubility, and swelling power all increased upon enzyme treatments, and their effects increased upon mixed enzyme treatment. For gelatinization characteristics of RVA, peak viscosity, final viscosity, breakdown, and total setback viscosity increased in rice flours treated with mixed enzymes. Especially, in steeping method with mixed enzyme treatment, pectinase 0.05% and cellulase 0.05% treatment was suitable for minimizing damaged starch and high fine particle distribution of rice flours compared with single enzyme treatment.

허혈-재관류 심근세포의 DNA에서 8-hydroxydeoxyguanosine 생성

  • 유효진;정명희;김명석;임정규
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.82-82
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    • 1993
  • 허혈-재관류손상 심근세포의 DNA에서 8-hydroxydeoxyguanosine (8-OHdG) 생성을 검토하였다. 흰쥐 적출심장의 Langendorff 관류 표본에서 대동맥 차단에 의한 60분 허혈후 산소가 포화된 Kredb-Henseleit용액으로 30분간 재관류 하므로서 허혈-재관류 손상을 유도하였다. 재관류 후 심근세포에서 DNA를 추출하고 HPLC(EC detector)를 이용하여 8-OHdG를 측정하였다. 실험결과 허혈-재관류 심근세포의 DNA에서 8-OHdG 함량이 증가하였으며 이는 $O_2$ 제거물질인 superoxide dismutase와 OH 제거물질인 mannitol에 의하여 방지되었다. Xanthine oxidase외 경쟁적 길항약인 allopurinol도 8-OHdG 생성을 억제하였으며 단백분해효소 억제제인 phenylsulfonylfluoride 그리고 관류액에서 칼슘의 제거 또한 허혈-재관류 심근 DNA의 생성을 방지하였다. 이상의 결과 허혈심근의 재관류시 8-OHdG 생성이 증가하며 이는 재관류 손상과 같은 산화성 심근손상을 평가하는 좋은 Index가 될 수 있을 것으로 여겨진다.

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Effects of Artemisia selengensis Methanol Extract on Ethanol-Induced Hepatotoxicity in Rat Liver (쑥(물쑥)추출물이 에탄올에 의한 흰쥐의 간 손상에 미치는 영향)

  • 김경수;이명렬
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.25 no.4
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    • pp.581-587
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    • 1996
  • This study was done to investigate the effects of Artemisia selengensis methanol extract on ethanol-induced hepatotoxicty in rat liver. Sprague-Dawley(SD) rats weighing about 150g were divided into the following 4 groups : control group(CON), Astemisia selengensis methanol extract administered group(ASE), ethanol adminstered group(ETH) and Artemisia selengenis methanol extract and ethanol administered group(ASA). Ethanol and Artemisia selengenis methanol extract were administered orally by 5m1/kg and 200mg/kg body weight per day for 6weeks, respectively. Body weight, daily food intake and percent liver weight per body weight were significantly changed by ethanol administration in comparison to control group. The activities of serum alanine aminotransferase(ALT), asparate aminotransferase(AST), and hepatic TBA-reactants increased by ethanol were decreased significantly by Artemisia selengensis methanol extract compared with ethanol group. It was also obseued that superoxide dismutase, catalase and glutathione peroxidase were not changed by Artemisia selengensis methanol extract, whereas hepatic xanthine oxidase activity was inhibitied by Artemisia selengensis methanol extract as compared to ethanol group. The glutathione contents in liver decreased by ethanol adminstration, but glutathione levels increased in ASA compared with ethanol group. These results suggest that Artemisia selengenis methanol extract have a possible protective effect on the ethanol-induced hepatotoxicity in rat liver.

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Effect of ALDH2 Enzyme Activity on the Level of 8-Hydroxydeoxyguanosine in Tissues Following Ethanol Exposure (ALDH2 효소의 활성이 알코올 섭취에 의한 8-hydroxydeoxyguanosine의 장기별 농도에 미치는 영향)

  • Zhang, Yan Wei;Choi, Sheung-Hee;Kim, Yun-Sik;Moon, Sun-In;Eom, Sang-Yong;Kim, Yong-Dae;Kim, Heon
    • Journal of Life Science
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    • v.18 no.8
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    • pp.1173-1176
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    • 2008
  • Individuals who regularly consume excessive quantities of alcohol are at a greater risk of developing various cancers such as esophageal, pharyngeal and lung cancers compared to normal populations if they are deficient in ALDH2 enzyme activity. We evaluated oxidative DNA damage in the liver, brain, and lung tissues of Aldh2 +/+ and Aldh2 -/- mice after they had been subjected to acute ethanol exposure. The 8-hydroxydeoxyguanosine (8-OHdG) level in each tissue was evaluated as a biomarker of oxidative DNA damage. The 8-OHdG level in the liver, brain, and lung tissues was significantly increased following ethanol treatment. In addition, the level of 8-OHdG in the liver and lung tissues was affected by ALDH2 enzyme activity. This result suggests that ALDH2-deficient individuals may be more susceptible than wild-type ALDH2 individuals to ethanol-mediated diseases, including cancer.