• 제목/요약/키워드: lactate dehydrogenase activity

검색결과 393건 처리시간 0.022초

Sulforhodamine B Assay to Determine Cytotoxicity of Vibrio vulnificus Against Human Intestinal Cells

  • Lee, Byung-Cheol;Choi, Sang-Ho;Kim, Tae-Sung
    • Journal of Microbiology and Biotechnology
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    • 제14권2호
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    • pp.350-355
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    • 2004
  • Sulforhodamine B (SRB) assay is a rapid, sensitive, and inexpensive method for measuring cell proliferation and chemosensitivity. However, the lactate dehydrogenase (LDH) release assay is generally used to measure cytototoxicity of infectious microorganisms against host cells. In this study, we investigated the possibility of applying the SRB assay to determine cytotoxicity for infectious microorganisms, and compared the results with those obtained by the LDH release assay. We used Vibrio vulnificus as a model of infectious microorganisms. The SRB assay showed that V vulnificus strongly induced cytotoxic activity against human intestinal cells, Caco-2 and INT-407 cells. The degree of cytotoxicity closely correlated with infection time and number ratios of V. vulnificus to intestinal cells (MOI, multiplicity of infection). Furthermore, cytotoxicity values obtained by SRB assay correlated well with results obtained by the LDH release assay, and both assays gave a linear response with respect to MOI Heat-inactivation of V. vulnificus for 35 min at $60^{\circ}C$ did not induce cytotoxic activity, indicating that viability of V. vulnificus is crucial for cytotoxic activity against intestinal cells. Although both assays are suitable as cytotoxicity endpoints, the SRB assay is recommended for measuring cytotoxicity of infectious microorganisms against host cells because of its significantly lower cost and more stable endpoint than the LDH release assay.

소환시 발생되는 아황산 가스가 흰쥐 혈청 중의 효소 활성도 및 지질 함량에 미치는 영향 (The Effect of Sulfur Dioxide by Burn out on Enzyme Activity and Lipid Content in Rats)

  • 김해동;김병학;하헌;조수열;윤수홍
    • Environmental Analysis Health and Toxicology
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    • 제12권3_4호
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    • pp.61-66
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    • 1997
  • Recently, incidences of pulmonary-related diseases are increasing and natural ecosystem is destroyed year by year owing to air pollutions by industrial noxious gases such as sulfur dioxide and carbon dioxide, etc. Sulfur dioxide produced by oxidation of sulfur compounds is water soluble and causes several kinds of pulmonary diseases, damaging on plants and animals. This study was undertaken to investigate effects of acute exposure of rats to sulfur dioxide using biochemical parameters. The sAST (aspartate transaminase) activity was not significantly affected but sALT (alanine transaminase), sLDH (lactate dehydrogenase) and sALP (alkaline phosphatase) activities were significantly increased by acute sulfur dioxide gas exposure. Also sulfur dioxide gas treatment significantly diminished serum cholinesterase activity. The total lipid, triglyceride, total-, HDL- and LDL-cholesterol contents were significantly increased by sulfur dioxide gas exposures. But the body and liver weight were generally diminished in comparison with control group. Our results' showed increased enzyme activities and lipid contents, but body and liver weight were all diminished by sulfur dioxide gas treatment.

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구속스트레스를 가한 흰쥐에서 Cholate류의 항스트레스 작용에 관한 연구 (Anti-stress Effect of Cholic acid Derivatives in Restraint Stress Induced Rats)

  • 박인;김양일;이선미;조태순
    • Biomolecules & Therapeutics
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    • 제4권2호
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    • pp.162-166
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    • 1996
  • This study was done to investigate whether cholic acid derivatives have anti-stress activity and what is a cause of this anti-stress effect. Seven cholic acid derivatives (cholic acid, taurocholic acid, ursodeoxycholic acid, tauroursodeoxychoic acid, chenodeoxy cholic acid, dehydrocholic acid, hyodeoxycholic acid) were used, silymarin and valproic acid were used as positive controls. Stress was induced by restraint immobilization technique plus water immersion (24hrs) and adrenal weight, spleen weight, adrenal ascorbic acid, serum cholesterol, lactate dehydrogenase (LDH), alkaline phosphatase (ALP), adrenal cholesterol, glucose and corticosterone levels were measured as stress indicators. Most cholic acid derivatives markedly decreased the adrenal weight, and TUDCA and DHCA increased the spleen weight. The restraint stress induced increments in serum LDH, ALP and cholesterol were attenuated by most cholic acid derivatives. Cholic acid, taurocholic acid and tauroursodeoxycholic acid only increased the content of adrenal ascorbate. While valproic acid showed an inhibitory effect against stress, silymarin did not. Our findings suggest that most cholic acid derivatives have anti-stress effect and that their anti-stress effect is, in part, related to choleretic activity.

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Construction of a Bile-responsive Expression System in Lactobacillus plantarum

  • Chae, Jong Pyo;Pajarillo, Edward Alain;Hwang, In-Chan;Kang, Dae-Kyung
    • 한국축산식품학회지
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    • 제39권1호
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    • pp.13-22
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    • 2019
  • This study aimed to develop a bile-responsive expression system for lactobacilli. The promoters of four genes, encoding phosphoenolpyruvate-dependent sugar phosphotransferase (mannose-specific), L-lactate dehydrogenase (LDH), HPr kinase, and D-alanine-D-alanine ligase, respectively, which were highly expressed by bile addition in Lactobacillus johnsonii PF01, were chosen. Each promoter was amplified by polymerase chain reaction and fused upstream of the ${\beta}$-glucuronidase gene as a reporter, respectively. Then, these constructs were cloned into E. coli-Lactobacillus shuttle vector pULP2, which was generated by the fusion of pUC19 with the L. plantarum plasmid pLP27. Finally, the constructed vectors were introduced into L. plantarum for a promoter activity assay. The LDH promoter showed the highest activity and its activity increased 1.8-fold by bile addition. The constructed vector maintained in L. plantarum until 80 generations without selection pressure. A bile-responsive expression vector, $pULP3-P_{LDH}$, for Lactobacillus spp. can be an effective tool for the bile-inducible expression of bioactive proteins in intestine after intake in the form of fermented dairy foods.

흰쥐에서 황칠나무 열수 추출물을 포함한 혼합물의 혈중 알코올 농도와 숙취 해소 효과 (Eliminatory Effect of Mixture including Hot Water Extract of Dendropanax morbifera Lev. on Alcohol-induced Blood Alcohol Concentration and Hangover in Rat)

  • 나주련;김은;박소이;이기훈;정의선;김진석;김용재;김선오
    • 한국키틴키토산학회지
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    • 제23권4호
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    • pp.267-276
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    • 2018
  • 본 연구에서는 숙취해소에 좋은 것으로 알려진 식품 소재와 황칠나무 추출물을 복합하여 in vitro 및 in vivo에서 알코올 유도된 간 손상에 대한 보호효과를 검토하였다. HepG2세포에서 300 mM 알코올과 SBJ 혼합물을 처리하였을 때 농도 의존적으로 LDH 수치가 감소하였으나 황칠나무 추출물을 제외한 복합물인 SBJ-복합물에서는 그 효과가 유의적으로 낮아지는 것을 관찰하였다. In vivo에서 SBJ 혼합물의 보호효과를 확인하고자 흰쥐에 알코올과 SBJ 혼합물을 투여한 결과, 알코올 투여 후 1시간까지 급격하게 혈중 알코올 수치가 EtOH군에서 증가하는 것이 관찰되었으며, SBJ 혼합물 투여군은 유의적인 차이로 감소되었음이 관찰되었으며 또한 농도의존적인 경향을 확인하였다. ADH 및 ALDH 활성의 증가는 SBJ 혼합물의 알코올 분해 및 대사산물의 제거 활성에 기여할 것으로 예상된다. 뿐만 아니라, 알코올에 의하여 증가한 LDH의 농도가 대조군과 유사한 수준으로 유지하는 것을 확인하였으며, GST, SOD, GPx 및 reduced glutathione와 같은 항산화 인자 및 효소의 활성은 대조군보다 EtOH군이 유의적으로 감소했으며, SBJ 혼합물에 의해 개선되는 경향을 관찰하였다. 이를 통해 SBJ 혼합물은 ADH, ALDH 활성 증가 및 항산화 방어계를 향상시킴으로써 산화적 스트레스 감소를 통한 간보호 효과가 있는 것으로 생각되며, 이러한 간보호에 미치는 주요한 추출물은 황칠나무 추출물임을 알 수 있었다. 이러한 결과를 토대로 향후 숙취해소 작용을 갖는 신규 식품소재로 활용될 수 있을 것으로 기대된다.

송화분이 Chloroform 투여 흰쥐의 혈청 및 간장에 미치는 영향 (Effect of Pine Pollen on the Chloroform Toxicity in Rat Serum and Liver)

  • 백경연;하은주;신해경;한준표
    • 동아시아식생활학회지
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    • 제10권4호
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    • pp.305-310
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    • 2000
  • This study was attempted to investigate the effect of pine pollen on the chloroform toxicity in rat serum and liver. The subjects were administrated with the graded concentraton of tow different levels of chloroform(1%, 5%) and an additional amount of pollen (1%, 5%) to some groups. the results were as follows: The activity of aspartate aminotrasferase(AST), alanine aminotrasferase(ALT) and lactate dehydrogenase(LDH) in serum, liver, and kidney increased in proportion to the chloroform concentration, but decreased in the pine pollen-treated groups. The amount of total cholesterol in serum of he chloroform administration groups was higher than that of the control group, and it decreased gradually with pine pollen administration.

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Antioxidant potential of silk protein sericin against hydrogen peroxide-induced oxidative stress in skin fibroblasts

  • Dash, Rupesh;Acharya, Chitrangada;Bindu, P.C.;Kundu, S.C.
    • BMB Reports
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    • 제41권3호
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    • pp.236-241
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    • 2008
  • The antioxidant potential of silk protein sericin from the non-mulberry tropical tasar silkworm Antheraea mylitta cocoon has been assessed and compared with that of the mulberry silkworm, Bombyx mori. Skin fibroblast cell line (AH927) challenged with hydrogen peroxide served as the positive control for the experiment. Our results showed that the sericin obtained from tasar cocoons offers protection against oxidative stress and cell viability is restored to that of control on pre-incubation with the sericin. Fibroblasts pre-incubated with non-mulberry sericin had significantly lower levels of catalase; lactate dehydrogenase and malondialdehyde activity when compared to untreated ones. This report indicates that the silk protein sericin from the non-mulberry tropical tasar silkworm, A. mylitta can serve as a valuable antioxidant.

Protective effect of green tea extract on doxorubicin induced cardiotoxicity in rats

  • Patil, Leena;Balaraman, R.
    • Advances in Traditional Medicine
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    • 제5권2호
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    • pp.137-143
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    • 2005
  • Doxorubicin induces oxidative stress leading to cardiotoxicity causing electrocardiogram abnormalities and increases in biomarkers associated with toxicity. Green tea extract (GTE) is reported to possess antioxidant activity mainly via its polyphenolic constituent, catechins. This study was intended to determine the effect of various doses of GTE (25, 50 and 100 mg/kg/day p.o. for 30 days) on doxorubicin-induced electrocardiographic and biochemical changes in rat heart. The latter included lactate dehydrogenase, creatine kinase, and glutamic oxaloacetate transaminase in serum and superoxide dismutase, catalase, and reduced glutathione, as well as membrane bound enzymes like $Na^+K^+ATPase,\;Ca^{2+}ATPase,\;Mg^{2+}ATPase$ and decreased lipid peroxidation in heart tissue Results demonstrated that rats which received GTE were less susceptible to such changes indicating protection afforded by GTE.

목초액의 혈소판 응집억제를 통한 혈행개선 작용에 관한 연구 (Improvement of Haemostasis Mediated by Anti-Platelet Activities by Plant Vinegar)

  • 김영대;배옥남;정승민;정진호
    • Toxicological Research
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    • 제20권2호
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    • pp.137-142
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    • 2004
  • We investigated the effects of plant vinegar on platelets and blood coagulation system. Plant vinegar inhibited in vitro platelet aggregation in a concentration dependent manner, when platelets were activated by thrombin and collagen. In addition, plant vinegar showed inhibitory effects on the serotonin secretion induced by thrombin in a concentration dependent manner. However, treatment with plant vinegar to platelets did not induce any cytotoxicity, as determined by the release of lactate dehydrogenase. Plant vinegar did not change the coagulation parameters such as activated partial thromboplastin time (aPTT) and prothrombin time (PT) using rat citrated plasma. In vivo study revealed that, treatment with plant vinegar prolonged the bleeding time from mouse tail. All these results suggest that plant vinegar might improve blood hemostasis mediated via anti platelet activities.

몰약 전탕액이 배양 심근세포에 미치는 영향 (Effects of Myrrha Water Extract on Rat Myocardial Cells in Cultures)

  • 권강범;조현익;김구환;김상범;이호섭;황우준;박승택;류도곤
    • 대한한의학회지
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    • 제21권2호
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    • pp.79-86
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    • 2000
  • Objectives and Methods : In order to elucidate toxic mechanism of myocardial damage and protective effect of myrrha water extract against cytotoxic effect of xanthine oxidase/hypoxanthine(XO/HX), cardioprotective effect of myrrha water extract was examined by MTT assay, LDH (Lactate Dehydrogenase) activity and heart beating rate after cultured myocardial cells derived from neonatal mouse were treated with various concentration of XO/HX, a free radical. Results : XO/HX induced a decrease of cell viability, an increase in the amount of LDH, and a decrease of heart beating rate on cultured myocardial cells in a dose-dependent manner. In cardioprotective effect of myrrha water extract, it showed a decrease in the amount of LDH and an increase of heart beating rate on cultured myocardial cells damaged by XO/HX. Conclusions : From the above results, it is suggested that XO/HX showed toxic effect in cultured myocardial cells derived from neonatal mouse and that myrrha water extract is very effective in the prevention of XO/HX-induced cardiotoxicity.

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