• Title/Summary/Keyword: LDH leakage

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The Cytotoxicity of Rat Hepatocytes on Viscum album Lectin (겨우살이 lectin의 흰쥐 간세포 독성에 미치는 영향)

  • Chang, Choul-Soo;Ryu, Jae-Ki;Kim, Tae-Ho
    • Korean Journal of Clinical Laboratory Science
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    • v.39 no.3
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    • pp.190-195
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    • 2007
  • In the present study, the author investigated to the cytotoxocity in cultured rat hepatocytes of Viscum album lectin. The cytotoxcity effect in Viscum album lectin on the activity of LDH was also investigated. Viscum album lectin significantly increased LDH leakage into medium of hepatocytes treated or untreated with $CCl_4$ (p<0.001). However, Viscum album lectin significantly increased LDH leakage from $CCl_4$-induced hepatocyte (p<0.001). There was a significant increase in LDH levels relative to the control group. Histological observation basically supported the result obtained from LDH assay. The livers of rats challenged with $CCl_4$ produced a marked increased cytoplasmic vacuoles and inflammatory cells in number, while the number of necrotic cells and swollen hepatocytes did not change significnatly. Rats administered DMSO alone did not alter the normal hepatic architecture. Histological observation of liver section in rat treated 72 hrs with either Viscum album lectin $CCl_4$-induced liver damage showed number of cytoplasmic vaculoe and necrotic cell. The number of inflammatory cell increased markedly. This results suggest to the conclusion that Viscum album lectin has a effect of hepatotoxicity activator.

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Effects of Extracellular Calcium and Starvation on Biochemical Indices of the Rat Hepatocytes

  • Kim, Ki-Sung
    • Toxicological Research
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    • v.11 no.2
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    • pp.199-203
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    • 1995
  • The focus of this study was to investigate that cellular parameters and glucose uptake might be altered by extracellular calcium and starvation. Addition of 1 mM $Ca^{++}$ to hepatocytes (equalling to the free calcium concentration of blood) significantly increased intracellular $Na^+$ and decreased $Na^+$ & LDH leakage. This pertains to the hepatocytes of control rats as well as those of rats fasted for 24 and 48. hr. These effects might be come from the membrane-stabilizing effects of calcium. But calcium had no effects on cell volumes, superoxide-formation and glucose uptake. Actually hepatocytes of starved rats showed changes in several cellular parameters. Starvation increased LDH leakage, glucose uptake and the total concentration of $Na^+$ and $Na^+$ whereas it markedly decreased cell volumes. Since total tonicity remained unchanged, intracellular $Na^+$ and $Na^+$ could contribute to a higher share of total osmolarity in starvation. Starvation increased the cytoplasmic pH because $R-NH^{3+}$ions and their corresponding counterions disappeared. This increase may be related to suppress the protonization of amino groups in proteins. Starvation decreased hepatic glycogen, a major compound that affects cytosolic volume of hepatocytes. The data indicate that starvation increases the glucose transport activity. The possible molecular basis will be discussed.

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Toxic Effect of Inhaled Toluene on the Neural Cell (톨루엔 흡입이 신경세포에 미치는 독성)

  • 김대병;류종훈;신대섭;이종권;정경자;류승렬;최기환;이선희;김부영
    • Toxicological Research
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    • v.13 no.3
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    • pp.251-256
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    • 1997
  • Toluene inhalation increases glutamate level and its receptor in various brain regions. In this study, nitric oxide synthase (NOS) activities were investigated in various rat brain regions using NADPH diaphorase staining method which examined histochemical changes of NOS in the neural cells. Also, in vitro LDH leakage assay and MTT test were performed to investigate the toxic influences of toluene in cultured granule cell of rat cerebellum which was significantly affected with toluene in vivo. Rats were exposed to toluene of 10000 ppm for 3 days. 7 days and 14 days by 20 min $\times$ 2 times a day. NADPH diaphorase staining was processed in the different brain regions after inhalation. NADPH diaphorase staining density was not significantly changed at 3 days inhalation group, but the density decreased in proportion to the duration of toluene inhalation. Over 30% of staining density was decreased at 14 days group which was maximum duration of inhalation in this study. The tendency of staining density decrease was significant in granule cell of cerebellum. Cell death by toluene exposure was observed in cultured cerebellar granule cell. $EC_{50}$ measured with LDH leakage assay and MTT test were 43 mM and 72 mM respectively.

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Comparison of the Two in Vitro Cytotoxicity Assays in Primary Cultured Rat Hepatocytes - the Neutral Red (NR) and Lactate Dehydrogenase (LDH) Tests (흰쥐의 배양 간세포를 이용한 세포독성시험에 있어서 뉴트랄레드 및 젖산 탈수소효소법의 비교)

  • Lee, Kyung-Tae;Seo, Seong-Hoon
    • Journal of Pharmaceutical Investigation
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    • v.25 no.4
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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.

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In vitro Alternatives to Skin Irritation Test

  • Shin, Dae-Sup;Kim, Dai-Byung;Ryu, Seung-Rel;Lee, Sun-Hee;Koh, Jae-Sook;Park, Won-Sae;Kim, Pu-Young
    • Biomolecules & Therapeutics
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    • v.3 no.3
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    • pp.242-244
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    • 1995
  • In vitro cell culture system has been proposed as a promising alternative model to in vivo skin irritation test. These studies were performed to screen the cytotoxicity effects of surfactants using normal human skin fibroblasts. Cell membrane integrity assessed by the leakage of lactate dehydrogenase (LDH) and mitochondrial integrity by MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromides reduction test were affected in a dose dependent manner. The irritation potential of surfactants to human skin patch test, and the changes of capillary permeability by rabbit intradermal safety test were assessed as in vivo methods. Our results suggest that LDH leakage assay and MTT reduction test using cultured human fibroblasts could be predictive for the irritancy of various surfactants in human, and LDH assay is superior correlated with in vivo test (r=0.886) to MTT test with in vivotest (r=0.757).

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Hepatoprotective Effects of Various Enzyme Hydrolysates from Oysters on Tacrine-Induced Toxicity in Human Hepatoma Cells (타크린으로 유발한 간세포 독성에 대한 효소별 굴 가수분해물의 보호 효과)

  • Park, Hye-Jin;Do, Hyung-Joo;Kim, Ok-Ju;Kim, Andre;Ha, Jong-Myung
    • Journal of Life Science
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    • v.22 no.1
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    • pp.117-125
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    • 2012
  • This study investigated the potential hepatoprotective benefits of Crassostrea gigas oyster hydrolysates. Oysters are known to have many biofunctional properties. In particular, oyster enzymatic hydrolysates produce substances with beneficial functions. The potential hepatoprotective effects of C. gigas hydrolysates against damage induced by tacrine were evaluated in vitro in HepG2 cells. Peptides were generated from C. gigas by enzymatic hydrolysis with Neutrase, Flavourzyme, or Protamex enzyme preparations. Tacrine treatment induced considerable cell damage in HepG2 cells, as shown by significant leakage of glutamic oxaloacetic transaminase (GOT) and lactate dehydrogenase (LDH). Cells treated with C. gigas hydrolysates showed an increased resistance to oxidative challenge compared to control cells, as revealed by higher cell survival against tacrine-induced hepatotoxicity. In addition, treatment with C. gigas hydrolysates reduced the leakage of GOT and LDH. These findings indicate that enzyme hydrolysates derived from C. gigas may be of benefit for developing hepatoprotective foods and drugs.

A Clinical Study on the Changes of Serum Enzymes after Cardiopulmonary Bypass (개심술환자의 심정지 시간에 따른 혈청효소치 변동에 관한 연구)

  • Lee, Jong-Uk;Yang, Gi-Min
    • Journal of Chest Surgery
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    • v.22 no.5
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    • pp.713-723
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    • 1989
  • In order to assess the correlation of the myocardial damage and the duration of cardiopulmonary bypass, measurement of creatine kinase [CK], lactate dehydrogenase [LDH], asparatate aminotransferase [AST], and MB band of CK [CK-MB] were carried out on the first, third, fifth, seventh, and ninth day in 44 patients following open heart surgery [POD 1,3,5,7,9]. And the patients were divided into three groups according to the duration of aortic cross clamp time [ACT]: Group I [ACT< 60 minutes. n=19], Group II [60 minutes < ACT< 90 minutes, n=7] and Group III[90 minutes > ACT, n=18]. 1. The leakage of CK in total patients increased to the highest level at POD 1, with rapid decrease and recovery at POD 7. The leakage of CK in Group III were greater than in Group I from POD 1 to POD 3 [P < 0.01]. The recovery time of CK level was shorter in Group I [POD 3] than in Group II and III [POD 7]. 2. The serum levels of LDH in total patients increased to the highest level at POD 1, with slow recovery until POD 9. The levels of LDH in Group III were higher than in Group I until POD 9 [P < 0.005]. The levels of LDH in Group I and II recovered but not in Group III. 3. The serum levels of AST in total patients increased to the highest level at POD 1, with rapid decrease and recovery at POD 7. The levels of AST in Group III were greater than in Group I from POD 1 to POD 5 [P < 0.05]. The recovery time of AST level was shorter in Group I and II [POD 5] than in Group III [POD 7]. 4. The positive cases for CK-MB in 36 patients were 22 [61.1 %] as a whole, 5[41.6%] in Group I, 4[57.1 %] in Group II, 13[76.4 %] in Group III at POD 1, and a case in each group at POD 3, and only a case in Group Ill at POD 5. It is concluded that the myocardial injury was closely related with the duration of cardiopulmonary bypass in open heart surgery.

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Intravascular Hemolysis after Prosthetic Valve Replacement (인공판막 치환수술후의 용혈)

  • Jang, Won-Chae;Lee, Gye-Yeong;Kim, Sang-Hyeong
    • Journal of Chest Surgery
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    • v.25 no.12
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    • pp.1556-1562
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    • 1992
  • Forty two consecutive patients who had had valve replacement with St. Jude Medical prosthesis were studied on a view point of intravascular hemolysis. Patients were consisted of 14 mitral valve replacement, and 7 aortic valve replacement, and 21 double, mitral and aortic, valve replacement. Serum LDH, indirect bilirubin, GOT, hemoglobin levels and ret-iculocyte count were pursued in postopeative 1st day, 3rd day, 7th day, 14th day and 21th day. Postoperatively, all patients were not detected paravalvular leakage on the ech-ocardiographical study. The patients with double valve replacement revealed higher levels of LDH on postopeative 14th day[P<0 05] than those with single valve replacement. Among the patients with single valve replacement, the patients with aortic valve replacement revealed slightly higher levels of entire postopeative data, but considered insignificant. There was correlation between the severity of hemolysis and the size of replaced aortic valve. In the postoperative LDH levels, the patients with small sized-aortic valve[less than 21mm in diameter] replacement revealed higher levels of postoperative 3rd day, 7th day and 14th day than those with large size[more than 23mm in diameter]. The patients with high level LDH of greater than 800 WU /L on postoperative 7th day were 61.9%[26 of 42]. The high LDH frequency of DVR was 71.4%[15 of 21], MVR 50.0%[7 of 14] and AVR, 57.1%[4 of 7]. The level of LDH declined gradualiy thereafter through postoperative 3 weeks. In conclusion, intravascular hemolysis after prosthetic valve replacement was dependent on position of valve replacement and size of valve. And this study supports the conventional valve selection and usage in our hospital. The patients with subclinical hemolysis after valve replacement should be placed on a close observation.

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Cytoprotective Effect of Zinc-Mediated Antioxidant Gene Expression on Cortisol-Induced Cytotoxicity (Cortisol 유발 세포독성에 대한 아연 관련 항산화 유전자 발현 증가에 의한 세포보호 효과)

  • Chung, Mi Ja;Kim, Sung Hyun;Hwang, In Min
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.5
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    • pp.649-656
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    • 2015
  • The protective effect of zinc against cortisol-induced cell injury was examined in rainbow trout gill epithelial cells. Cells exposed to cortisol for 24 h showed increased leakage of lactate dehydrogenase (LDH) as well as decreased cell viability in a dose-dependent manner. Treatment with zinc ($100{\mu}M$ $ZnSO_4$) reduced the severity of both LDH release and cell death as well as protected cells against cortisol-induced caspase-3 activation, indicating reduction of apoptosis. Cortisol-induced cell death, leakage of LDH, and caspase-3 activation were blocked by the glucocorticoid receptor antagonist Mifepristone (RU-486), suggesting that cell injury was cortisol-dependent. In addition, we studied the effect of zinc on the expression of antioxidant genes such as metallothionein A (MTA), metallothionein B (MTB), glutathione-S-transferase (GST), and glucose-6-phosphate dehydrogenase (G6PD) during cortisol-induced cell injury. MTA, MTB, GST, and G6PD mRNA levels increased after treatment with zinc or cortisol, separately or in combination. Higher mRNA levels of MTA, MTB, GST, and G6PD were detected when cells were treated with $100{\mu}M$ $ZnSO_4$ and $1{\mu}M$ cortisol in combination at the same time compared to treatment with zinc or cortisol separately. Cells treated with zinc showed increased intracellular free zinc concentrations, and this response was significantly enhanced in cells treated with cortisol and zinc. In conclusion, zinc treatment inhibited cortisol-induced cytotoxicity and apoptosis through indirect antioxidant action.

Cytoprotective Effects of Natural Flavonoids on Carbon Tetrachloride-Induced Toxicity in Primary Cultures of Rat Hepatocytes (사염화탄소로 유도한 일차 배양 간세포 독성에서 Flavonoid류의 세포보호 효과)

  • Kim, Young-Kwan;Kim, Yang-Hee;Kim, Dong-Hyun;Lee, Kyung-Tae
    • Korean Journal of Pharmacognosy
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    • v.36 no.3 s.142
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    • pp.224-228
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    • 2005
  • Protective effects of various natural flavonoids on carbon tetrachloride $(CCl_4)-induced$ hepatotoxicity were investigated in primary cultured rat hepatocytes. Some of these flavonoids decreased the ALT and LDH releases induced by $CCl_4$ in A dose-dependent manner. Neohesperidin, hesperetin, baicalin, baicalein and quercetin inhibited $CCl_4-induced$ alanine aminotransferase (ALT) release. In addition, quercetin, quercitrin, neohesperidin, baicalin, baicalein and naringin reduced $CCl_4$ induced lactate dehydrogenase (LDH) leakage. Among these flavonoids, quercitrin, quercetin, baicalin and baicalein possessed potent protective effects and were selected for the further investigation on lipid peroxidation. These four flavonoids inhibited dose dependently $CCl_4-induced$ lipid peroxidation. Especially, the protective effects of quercetin and baicalein were similar to silybin as a well-known hepatoprotective agent. These results suggest that these four flavonoids have significant cytoprotective effects and possibility of therapeutic effect on chemical-induced liver diseases.