• Title/Summary/Keyword: LDH release

검색결과 250건 처리시간 0.032초

감초 추출물이 glutathione S-transferase의 유도 활성에 미치는 영향 (Induction of Glutathione S-transferase Activity by the Extracts of Glycyrrhiza uralensis Fischer)

  • 윤미영;전경임;손은순;김지현;김용성;박은주;박해룡
    • Applied Biological Chemistry
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    • 제51권3호
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    • pp.228-232
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    • 2008
  • 본 연구에서는 항산화 황성을 가지면서 신경세포 보호 효과를 가지고 있는 석곡, 형개, 감초 및 작약 추출물을 대상으로 GST 유도 활성 효과를 실험하였다. 그 결과, 감초 추출물이 가장 높은 GST 유도 활성을 나타내었다. 감초를 methanol, ethanol, acetone으로 분획 추출한 뒤 GST 유도 활성을 살펴본 결과, 감초의 methanol 추출물을 50 ${\mu}g/ml$ 농도로 처리 시 대조구에 비해 2.23배로 가장 논은 GTS 유도환성이 나타났다. 그리고 감초의 각 추출물들은 처리 농도에서 세포의 형태학적인 변화를 유도하지 않았으며, 이는 LDH release assay 결과를 통해서도 매우 낮은 세포 독성을 확인할 수 있었다. 감초의 GST 유도 활성 물질들의 용매특성을 알아보기 위하여 methanol 추출물을 극성도에 따라 용매 분획하여 GST 유도 활성을 확인한 결과, diethyl ether층 분획물에서 가장 높은 GST 유도 활성을 나타냄을 확인하였다.

Protective Effect of Schizonepeta tenuifolia Briquet Extracts on Oxidative DNA Damage in Human Leucocytes and on Hydrogen Peroxide-induced Cytotoxicity in PC12 Cells

  • Yoon, Mi-Young;Lee, Hyun-Jin;Lee, Bo-Bae;Lee, Sang-Myeon;Kim, Ju-Young;Kim, Yong-Seong;Park, Eun-Ju;Park, Hae-Ryong
    • Food Science and Biotechnology
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    • 제16권5호
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    • pp.858-862
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    • 2007
  • The present study was conducted to examine the antioxidant activities and neuroprotective effects of methanolic extracts from Schizonepeta tenuifolia Briquet (STE). STE ($100\;{\mu}g/mL$) showed $43.33\;{\mu}M$ of total phenolic content, 64.43% of radical scavenging activity, and 0.157 of reducing power. In addition, the effect of STE on $H_2O_2$-induced DNA damage in human leucocytes was evaluated by the comet assay, where STE was a dose dependent inhibitor of DNA damage induced by $200{\mu}M$ of $H_2O_2$. The protective effect of STE against $H_2O_2$-induced oxidative damage on PC12 cells was investigated by an 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) reduction assay and lactate dehydrogenase (LDH) release assays. After 2 hr of cell exposure to $H_2O_2\;(500\;{\mu}M)$, a marked reduction in cell survival was observed. However, this reduction was significantly prevented by $1-50\;{\mu}g/mL$ of STE. Therefore, these results suggest that STE could be a new antioxidant candidate against neuronal diseases.

Combination Doxorubicin and Interferon-α Therapy Stimulates Immunogenicity of Murine Pancreatic Cancer Panc02 Cells via Up-regulation of NKG2D ligands and MHC Class I

  • Wang, Wen-Jia;Qin, Si-Hao;Zhang, Ji-Wei;Jiang, Yue-Yao;Zhang, Jin-Nan;Zhao, Lei
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권22호
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    • pp.9667-9672
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    • 2014
  • Background: Pancreatic adenocarcinoma is a malignant gastrointestinal cancer with significant morbidity and mortality. Despite severe side effects of chemotherapy, the use of immunotherapy combined with chemotherapy has emerged as a common clinical treatment. In this study, we investigated the efficacy of the combined doxorubicin and interferon-${\alpha}$ (IFN-${\alpha}$) therapy on murine pancreatic cancer Panc02 cells in vitro and in vivo and underlying mechanisms. Materials and Methods: A Panc02-bearing mouse model was established to determine whether doxorubicin and interferon-${\alpha}$ (IFN-${\alpha}$) could effectively inhibit tumor growth in vivo. Cytotoxicity of natural killer (NK) cells and cytotoxic T lymphocytes (CTLs) was evaluated using a standard LDH release assay. To evaluate the relevance of NK cells and CD8 T cells to the combination therapy-mediated anti-tumor effects, they were depleted in tumor-bearing mice by injecting anti-asialo-GM-1 antibodies or anti-CD8 antibodies, respectively. Finally, the influence of doxorubicin+interferon-${\alpha}$ (IFN-${\alpha}$) on the ligands of NK and T cells was assessed by flow cytometry. Results: The combination therapy group demonstrated a significant inhibition of growth of Panc02 in vivo, resulting from activated cytotoxicity of NK cells and CTLs. Depleting CD8 T cells or NK cells reduced the anticancer effects mediated by immunochemotherapy. Furthermore, the doxorubicin+IFN-a treatment increased the expression of major histocompatibility complex class I (MHC I) and NKG2D ligands on Panc02 cells, suggesting that the combined therapy may be a potential strategy for enhancing immunogenicity of tumors. All these data indicate that the combination therapy using doxorubicin and interferon-${\alpha}$ (IFN-${\alpha}$) may be a potential strategy for treating pancreatic adenocarcinoma.

HPV16 CTL Epitope Peptide-activated Dendritic Cell and Natural Killer Co-culture for Therapy of Cervical Cancer in an Animal Model

  • Hu, Yan-Xia;Li, Min;Jia, Xiao-Hui;Du, Qu-Xiao;Miao, Feng-Tai;Yao, Li;Shen, Ji-Duo
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권12호
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    • pp.7335-7338
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    • 2013
  • There is increasing evidence that natural killer (NK) cells play an important role in antitumor immunity following dendritic cell (DC) vaccination. Little is known, however, about the optimal stimulation of DCs by epitopes and NK interactions for cytotoxicity in tumors. In this study, DC cells activated by the HPV16E7.49-57 epitope and LPS were co-cultured with NK cells in vitro, and then used ot immunize mice to study CTL activity of TC-1, which constitutively expresses HPV16E6E7, with an LDH release assay. Cytotoxicity in mice immunized with DC loaded with epitope HPVE7.49-57 vaccine co-cultured with NK was enhanced significantly (p<0.01). In conclusion, talk-across between DC and NK cells enhances their functions, also improving cytotoxicity againsttumor cells, suggesting that activated DC-NK by epitopes has potential application for cancer-specific immuno-cellular therapy.

홍화자약침액(紅花子藥鍼液)이 수은(水銀)에 의한 가토(家兎)의 신세뇨관(腎細尿管) 물질이동(物質移動) 변화(變化)에 미치는 영향(影響) (Effect of Carthami Semen Aquacupunture(CSA) on Mercury-Induced Alterations in Tubular Transport Function in Rabbits)

  • 최영규;윤현민;송춘호;장경전;안창범
    • Journal of Acupuncture Research
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    • 제19권5호
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    • pp.199-208
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    • 2002
  • Objective : This study was undertaken to determine if Carthami Semen Aquacupunc- ture(CSA) exerts protective effect against alterations in membrane transport function rabbits with mercury chloride(HG)-induced acute renal failure. Methods : The administration of Hg at a subcutaneous single dose of 10 mg/kg caused a reduction in GFR and an increase in fractional Na excretion, indicating generation of acute renal failure. When CSA were given for 7 days prior to Hg administration, such changes were significantly attenuated. The fractional excretion of glucose and phosphate was increased in rabbits treated with Hg alone. Results : The increase in rabbits treated with Hg following CSA are significantly lower than that in animals treated with Hg alone. Uptakes of glucose and phosphate in purified isolated brush-border membrane and Na-K-ATPase activity in microsomal fraction were inhibited in rabbits treated with Hg alone. Such changes were prevented by CSA. Uptakes of organic ions, PAH and TEA, in renal cortical slices were inhibited by the administration of Hg, which was prevented by CSA. Exposure of renal cortical slices to Hg in vitro caused an increased LDH release and lipid peroxidation, which was significantly prevented by CSA extract. Conclusions : These results indicate that the administration of Hg causes impairment in reabsorption of solutes in the proximal tubule via the generation of reactive oxygen species. CSA provides the protection against the impairment in proximal reabsorption, and its effect may be resulted from its antioxidant effect.

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Protective Effect of Polysaccharide Fractions from Radix A. Sinensis against tert-Butylhydroperoxide Induced Oxidative Injury in Murine Peritoneal Macrophages

  • Yang, Xingbin;Zhao, Yan;Lv, You;Yang, Ying;Ruan, Yun
    • BMB Reports
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    • 제40권6호
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    • pp.928-935
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    • 2007
  • Three Angelica sinensis polysaccharide fractions (APFs), named APF1, APF2 and APF3, were isolated and purified from Radix A. sinensis and their antioxidant activities were evaluated in isolated mouse peritoneal macrophages by pretreatment with APFs before exposure to 0.2 mM tertbutylhydroperoxide (t-BHP). The results showed that pretreatment of the macrophages with APFs as low as $10{\mu}g$/ml could significantly enhance t-BHP-decreased cell survival, intracellular glutathione (GSH) content and superoxide dismutase (SOD) activity, and also inhibited t-BHP-increased lactate dehydrogenase (LDH) leakage and malondialdehyde (MDA) formation (p < 0.05), and APF3 was the most active fraction, followed by APF2 and APF1 in decreasing order. Furthermore, we found for the first time that the bound-protein in APF3 was associated closely with the protective effects and the polysaccharide inhibited the excess NO release from t-BHP-activated macrophages to protect host cells.

사물탕이 산화적 스트레스에 의하여 유발되는 신경세포의 세포 사멸에 미치는 보호효과 (Protective Effects of Samul-tang on Cell Death Inducded by Oxidative Stress in C6 Glial Cell)

  • 김형우;김경윤;김계엽;김재현;정종길;최찬헌;황귀성;이상영;정현우
    • 동의생리병리학회지
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    • 제23권5호
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    • pp.969-973
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    • 2009
  • Samul-tang (SMT), which was firstly described in (Hwajegukbang) Song dynasty, is well known remedy for blood diseases in Oriental medicine. SMT is traditional herbal-remedy composed of Rehmanniae Radix Preparat, Angelicae Gigantis Radix, Cnidii Rhizoma and Paeoniae Radix. Recently, SMT has known to have anti-oxidative action. However, the reports on anti-oxidantic action in neuroglial cells are rare. In addition, the exact mechanisms are unclear. For these reasons, we investigated the protective effects of SMT on cell death induced by oxidative stress using C6 glioma cells. In our results, SMT accelerated proliferation rates of C6 cells in vitro. In addition, levels of LDH release induced by oxidative stress were lowered by treatment with SMT. Finally, protective effects on cell death induced by chemicals such as paraquat and rotenone were observed. In conclusion, these results suggest the possibility to protect brain cell or neuronal cell from damage induced by oxidative stress.

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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Fucoidan Attenuates Perfluorooctane Sulfonate-induced Apoptosis of Neuronal Cells

  • Lee, Youn Ju;Sim, Kyung-Hwa;Yang, Jae-Ho
    • 한국해양바이오학회지
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    • 제9권1호
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    • pp.1-7
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    • 2017
  • Perfluorooctane sulfonate (PFOS) is one of the most widely distributed environmental pollutants and causes neurotoxicities. Fucoidan is a main bioactive constituent of the brown sea-weed and has many functions in a variety of physiological conditions. The present study attempted to investigate the potential role of fucoidan as neuroprotective marine polypeptide in environmental pollutant-induced apoptosis of neuronal cells in culture. MTT assay showed that cell viability was significantly reduced to 68 % at $30{\mu}M$ PFOS, which was recovered up to 77% and 92% in the presence of fucoidan 25 and $50{\mu}g/ml$, respectively. Cytotoxicity assay showed that LDH release was significantly increased to 160% at $30{\mu}M$ PFOS but was reduced to 150% and 122% in the presence of fucoidan 25 and $50{\mu}g/ml$, respectively. Caspase-3 activity, a hallmark of apoptosis, was measured to determine the cytotoxicity of PFOS and the cytoprotective effects of fucoidan. PFOS induced a 250% increase of caspase-3 activity at $30{\mu}M$ but the increase was dampened to 180% and 130% in the presence of fucoidan 25 and $50{\mu}g/ml$, respectively. PFOS $30{\mu}M$ induced 180 % increase in ROS accumulation, which was effectively blocked by $50{\mu}g/ml$ fucoidan (120% of control). Our results demonstrated that PFOS is a powerful neurotoxicant and fucoidan may be a protective marine bioactive polypeptide against the neurotoxic environmental pollutants. It may contribute to establishing the potential role of fucoidan as a neuroprotective polypeptide that prevents the risk of neurological disorders from the possible neurotoxic pollutants.

JAK-2 V617F Mutational Analysis in Primary Idiopathic Myelofibrosis: Experience from Southern Pakistan

  • Sultan, Sadia;Irfan, Syed Mohammed
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권17호
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    • pp.7889-7892
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    • 2015
  • Background: Primary idiopathic myelofibrosis (PMF) is a clonal Ph-chromosome negative myeloproliferative neoplasm characterized by dysregulated kinase signaling and release of abnormal cytokines. In the recent past, following JAK2 V617F mutation invention, important revolution has been made in the molecular diagnostic biology of this disease. The rational of this study was to determine the mutational status of JAK2 V617F in Pakistan patients with PMF. Materials and Methods: In this cross sectional study, 20 patients with PMF were enrolled from January 2011 to December 2014. Diagnosis was based on WHO criteria for PMF. All patients were screened for G-T point mutation (V617F) in the JAK2 gene on chromosome 9 by allele specific PCR. Results: The mean age was $57.9{\pm}16.5years$. The male to female ratio was 3:1. The frequency of JAK2 V617F positivity in our PMF patients was found to be 55%. Positive correlations of JAK2 V617F mutation were established with high TLC count, raised LDH and marked splenomegaly (P<0.05). No correlation of JAK2 V617F could be established with age and gender (P>0.05). Conclusions: The JAK2 V617F mutation frequency in our PMF patients was similar to those reported previously. In our hands JAK2 V617F mutated patients expressed an aggressive disease phenotype. Screening for the mutation in all suspected PMF cases could be beneficial in differentiating patients with reactive and clonal marrow fibrosis.