• 제목/요약/키워드: Serum deprivation

검색결과 57건 처리시간 0.029초

Korean Red Ginseng protects endothelial cells from serum-deprived apoptosis by regulating Bcl-2 family protein dynamics and caspase S-nitrosylation

  • Kim, Young-Mi;Kim, Jung Hwan;Kwon, Hyuk Min;Lee, Dong Heon;Won, Moo-Ho;Kwon, Young-Guen;Kim, Young-Myeong
    • Journal of Ginseng Research
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    • 제37권4호
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    • pp.413-424
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    • 2013
  • Korean Red Ginseng extract (KRGE) is a traditional herbal medicine utilized to prevent endothelium dysfunction in the cardiovascular system; however, its underlying mechanism has not been clearly elucidated. We here examined the pharmacological effect and molecular mechanism of KRGE on apoptosis of human umbilical vein endothelial cells (HUVECs) in a serum-deprived apoptosis model. KRGE protected HUVECs from serum-deprived apoptosis by inhibiting mitochondrial cytochrome c release and caspase-9/-3 activation. This protective effect was significantly higher than that of American ginseng extract. KRGE treatment increased antiapoptotic Bcl-2 and Bcl-$X_L$ protein expression and Akt-dependent Bad phosphorylation. Moreover, KRGE prevented serum deprivation-induced subcellular redistribution of these proteins between the mitochondrion and the cytosol, resulting in suppression of mitochondrial cytochrome c release. In addition, KRGE increased nitric oxide (NO) production via Akt-dependent activation of endothelial NO synthase (eNOS), as well as inhibited caspase-9/-3 activities. These increases were reversed by co-treatment of cells with inhibitors of eNOS and phosphoinositide 3-kinase (PI3K) and pre-incubation of cell lysates in dithiothreitol, indicating KRGE induces NO-mediated caspase modification. Indeed, KRGE inhibited caspase-3 activity via S-nitrosylation. These findings suggest that KRGE prevents serum deprivation-induced HUVEC apoptosis via increased Bcl-2 and Bcl-$X_L$ protein expression, PI3K/Akt-dependent Bad phosphorylation, and eNOS/NO-mediated S-nitrosylation of caspases. The cytoprotective property of KRGE may be valuable for developing new pharmaceutical means that limit endothelial cell death induced during the pathogenesis of vascular diseases.

PHYSIOLOGICAL RESPONSES, FEED INTAKE, URINE VOLUME AND SERUM OSMOLALITY OF AARDI GOATS DEPRIVED OF WATER DURING SPRING AND SUMMER

  • El-Nouty, F.D.;Al-Haidary, A.A.;Basmaeil, S.M.
    • Asian-Australasian Journal of Animal Sciences
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    • 제3권4호
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    • pp.331-336
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    • 1990
  • In order to evaluate the adaptability of Aardi goats to arid environment, 5 Aardi bucks were deprived of water for four days during spring and summer seasons. The rise in average maximum ambient temperature from $24.8^{\circ}C$ in spring to $35.8^{\circ}C$ in summer caused a significant rise in rectal temperature ($0.3^{\circ}C$), respiratory rate (62%), serum osmolaity (8%) and serum sodium concentration (17%). While, it resulted in a significant decline in dry matter intake (50%), urine volume (74%) and fecal water excretion (60%) compared with their values in spring, but had no significant effect on the volume of drinking water. At the end of the 4-days deprivation period during spring, respiratory rate, dry matter intake and urine volume were reduced by 18, 77 and 91% relative to their average in control goats. The corresponding reduction in summer were 58, 100 and 100%. Serum osmolaity was risen by 15% in spring deprived goats and 29% in summer deprived goats. Rectal temperature rose by a mean value of $1.3^{\circ}C$ only in goats deprived of water in summer. Percent of moisture in the feces declined from 64 in control goats, to 24% in water deprived goats during spring season. The corresponding values in summer were 25 and 6%. These responses of Aardi goats deprived of water in summer indicate that they possess a water economy mechanism enable them to tolerate infrequent drinking in hot-arid environment.

Role of Protein Kinases on NE-$_{\kappa}B$ Activation and Cell Death in Bovine Cerebral Endothelial Cells

  • Ahn, Young-Soo;Kim, Chul-Hoon;Kim, Joo-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • 제3권1호
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    • pp.11-18
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    • 1999
  • Nuclear factor $_{\kappa}B\;(NF-_{\kappa}B)$ activation is modulated by various protein kinases. Activation of $NF-_{\kappa}B$ is known to be important in the regulation of cell viability. The present study investigated the effect of inhibitors of protein tyrosine kinase (PTK), protein kinase C (PKC) and protein kinase A (PKA) on $NF-_{\kappa}B$ activity and the viability of bovine cerebral endothelial cells (BCECs). In serum-deprivation-induced BCEC death, low doses of $TNF{\alpha}$ showed a protective effect. $TNF{\alpha}$ induced $NF-_{\kappa}B$ activation within 4 h in serum-deprivation. PTK inhibitors (herbimycin A and genistein) and PKC inhibitor (calphostin C) prevented $NF-_{\kappa}B$ activation stimulated by $TNF{\alpha}.$ Likewise, these inhibitors prevented the protective effect of $TNF{\alpha}.$ In contrast to $TNF{\alpha}-stimulated\;NF-_{\kappa}B$ activity, basal $NF-_{\kappa}B$ activity of BCECs in media containing serum was suppressed only by calphostin C, but not by herbimycin A. As well BCEC death was also induced only by calphostin C in serum-condition. H 89, a PKA inhibitor, did not affect the basal and $TNF{\alpha}-stimulated\;NF-_{\kappa}B$ activities and the protective effect of $TNF{\alpha}$ on cell death. These data suggest that modulation of $NF-_{\kappa}B$ activation could be a possible mechanism for regulating cell viability by protein kinases in BCECs.

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Ebb-and-Flow of Macroautophagy and Chaperone-Mediated Autophagy in Raji Cells Induced by Starvation and Arsenic Trioxide

  • Li, Cai-Li;Wei, Hu-Lai;Chen, Jing;Wang, Bei;Xie, Bei;Fan, Lin-Lan;Li, Lin-Jing
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권14호
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    • pp.5715-5719
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    • 2014
  • Autophagy is crucial in the maintenance of homeostasis and regenerated energy of mammalian cells. Macroautophagy and chaperone-mediated autophagy(CMA) are the two best-identified pathways. Recent research has found that in normal cells, decline of macroautophagy is appropriately parallel with activation of CMA. However, whether it is also true in cancer cells has been poorly studied. Here we focused on cross-talk and conversion between macroautophagy and CMA in cultured Burkitt lymphoma Raji cells when facing serum deprivation and exposure to a toxic compound, arsenic trioxide. The results showed that both macroautophagy and CMA were activated sequentially instead of simultaneously in starvation-induced Raji cells, and macroautophagy was quickly activated and peaked during the first hours of nutrition deprivation, and then gradually decreased to near baseline. With nutrient deprivation persisted, CMA progressively increased along with the decline of macroautophagy. On the other hand, in arsenic trioxide-treated Raji cells, macroautophagy activity was also significantly increased, but CMA activity was not rapidly enhanced until macroautophagy was inhibited by 3-methyladenine, an inhibitor. Together, we conclude that cancer cells exhibit differential responses to diverse stressor-induced damage by autophagy. The sequential switch of the first-aider macroautophagy to the homeostasis-stabilizer CMA, whether active or passive, might be conducive to the adaption of cancer cells to miscellaneous intracellular or extracellular stressors. These findings must be helpful to understand the characteristics, compensatory mechanisms and answer modes of different autophagic pathways in cancer cells, which might be very important and promising to the development of potential targeting interventions for cancer therapies via regulation of autophagic pathways.

Differential Proteome Expression of in vitro Proliferating Hanwoo Stromal Vascular Cells from Omental, Subcutaneous and Intramuscular Depots in Response to Hormone Deprivation and IGF-1, Estradiol-17β Addition

  • Rajesh, Ramanna Valmiki;Kim, Seong-Kon;Park, Min-Ah;Kwon, Seulemina;Chang, Jong-Soo;Yoon, Du-Hak;Kim, Tae-Hun;Lee, Hyun-Jeong
    • Journal of Animal Science and Technology
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    • 제52권3호
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    • pp.175-186
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    • 2010
  • The aim of this study was to analyze the proteome expressions of proliferating stromal vascular cells from Hanwoo omental, subcutaneous and intramuscular depots subjected to hormone deprivation and IGF-1, Estradiol-$17{\beta}$ addition. For hormone deprivation or addition studies, the cells were either grown in 10% charcoal-dextran stripped fetal bovine serum (CD-FBS) or in 10% FBS supplemented medium. Further, to analyze the effect of insulin like growth factor (IGF-1) and $17\beta$-Estradiol (E2), cells were grown in 10% CD-FBS containing IGF-1 (10 ng/ml) or E2 (10 nM). The results showed that hormone deprivation had a negative impact on proliferation among the cells from all depots without any growth difference. On comparison of proliferation levels, higher levels were observed in cells that were grown in 10% FBS than in 10% CD-FBS alone or with IGF-1/E2. Proteome expression from preadipocytes grown in hormone deprivation conditions were compared by 2D-DIGE and MALDIToF/ToF. A total of twelve different proteins were found to be differentially expressed under hormone deprivation conditions. Further, our proteomic analysis with DIGE under IGF-1 and E2 addition revealed four proteins with differential expression levels. Moreover, the results highlighted in this study offer a role for each differentially expressed protein with respect to their effect in positive or negative regulation on proliferation.

Differential Proteome Expression of In vitro Proliferating Bovine Satellite Cells from Longissimus Dorsi, Deep Pectoral and Semitendinosus Muscle Depots in Response to Hormone Deprivation and Addition

  • Rajesh, Ramanna Valmiki;Kim, Seong-Kon;Park, Mi-Rim;Park, Min-Ah;Jang, Eun-Joung;Hong, Seung-Gu;Chang, Jong-Soo;Yoon, Du-Hak;Kim, Tae-Hun;Lee, Hyun-Jeong
    • Journal of Animal Science and Technology
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    • 제51권6호
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    • pp.459-470
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    • 2009
  • The aim of this study was to analyze the proteome of proliferating bovine satellite cells from longissimus dorsi, deep pectoral and semitendinosus muscle depots which had been subjected to hormonal deprivation or addition in culture. For hormone deprivation or addition studies, the cells were either grown in 10% charcoal-dextran stripped fetal bovine serum (CD-FBS) or in 10% FBS supplemented medium. Further to analyze the effect of insulin like growth factor (IGF-1) and testosterone (TS), the cells were grown in 10% CD-FBS containing IGF-1 (10 ng/ml) or TS (10 nM). Results have shown that hormone deprivation had a negative impact on proliferation of the cells from each of the muscle depots. In case of IGF-1 and TS addition, the proliferation levels were low compared with that of the cells grown in 10% FBS. Hence, to gain the insights of the proteins that are involved in such divergent levels of proliferation, the proteome of such satellite cells proliferating under the above mentioned conditions were analyzed using 2D-DIGE and MALDI-ToF/ToF. Thirteen proteins during hormone deprivation and nine proteins from hormone addition were found to be differentially expressed in all the cultures of the cells from the three depots. Moreover, the results highlighted in this study offer a role for each differentially expressed protein with respect to its effect on positive or negative regulation of cell proliferation.

단미 한약 복용중 발견한 Gilbert's syndrome 2예 임상고찰 (Report on Two Cases of Gilbert's Syndrome Found in the Process of Administering Herbs)

  • 이종덕;김동웅
    • 동의생리병리학회지
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    • 제28권6호
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    • pp.657-661
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    • 2014
  • Gilbert's syndrome is one that shows a benign course with intermittent unconjugate hyperbilirubinemia without any evidence of hepatobiliary tract disease or hemolysis. It is often found in a health examination or blood laboratory test by chance. In particular, patients who are taking drugs, including herbal medicine should be careful for their medication due to the possibility of associations with changes in liver function because of drug metabolism, sometimes they have to quit the use of the medication for a certain period and often they should get an additional test. Two male patients increased serum total bilirubin level without other systemic symptoms in screening test for clinical herb medicine pharmacokinetics study. Therefor they was diagnosed with suspected Gilbert's syndrome. They had been calory deprivation test with 24 hours fasting state. They also performed liver function test and ultrasonogram for evaluation of hepatobiliary tract disease. Total serum bilirubin was markedly increased, especially unconjugate bilirubin level higher over the two times than base line after they had been calory deprivation for 24 hours, They was not found another abnormality all laboratory results and physical examination. This study is a report on two cases of hyperbilirubinemia, diagnosed as Gilbert's syndrome, which were found in the process of a clinical pharmacokinetic study of a decoction of medicinal herbs.

허혈 상태의 PC12 세포에 대한 대보원전(大補元煎)의 신경보호효과 (Neuroprotective Effects of Daebowonjeon on PC12 Cells Exposed to Ischemia)

  • 김봉상;이선우;문병순
    • 동의생리병리학회지
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    • 제21권1호
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    • pp.117-125
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    • 2007
  • Neuronal ischemia is a pathological process caused by a lack of oxygen (anoxia) and glucose (hypoglycemia), resulting in neuronal death. It is believed that apoptosis is one of the mechanisms involved in ischemic cell death. Neuronal apoptosis is a process characterized by nuclear DNA fragmentation, changes of plasma membrane organization. To elucidate the mechanism of neuronal death following ischemic insult and to develop neuroprotective effects of Daebowonjeon(DBWJ) against ischemic damage, in vitro models are used. In vitro models of cell death have been devloped with pheochromocytoma (PC12) cell, which have become widely used as neuronal models of oxidative stress, trophic factor, serum deprivation and chemical hypoxia. Using a special ischemic device and PC12 cultures, we investigated an in vitro model of ischemia based on combined Oxygen and Glucose Deprivation (OGD) insult, followed by reoxygenation, mimicking the pathological conditions of ischemia. In this study, Daebowonjeon rescued PC12 cells from Oxygen-Glucose Deprivation (OGD)-induced cell death in a dose-dependent manner The nuclear staining of PC12 cells clearly showed that DBWJ attenuated nuclear condensation and fragmentation which represent typical neuronal apoptotic characteristics. DBWJ also prevents the LDH release and induction of Hypoxia Inducing Factor (HIF)-1 by OGD-exposed PC12 cells. Furthermore, DBWJ reduced the activation of polyADP-ribose polymerase (PARP) by OGO-exposed PC12 cells. These results suggest that apoptosis is an important characteristic of OGD-induced neuronal death and that oriental medicine, such as DBWJ, may prevent PC12 cell from OG D-induced neuronal death by inhibiting the apoptotic process.

감맥대조탕 복합추출물의 만성스트레스 유발 생쥐의 항우울 작용에 관한 실험연구 (Experimental Study on the Anti-Depressant Effects of Gammakdaejo-tang Complex Extracts in Rats Induced with Chronic Mild Stress)

  • 이유찬;김보경
    • 동의신경정신과학회지
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    • 제29권1호
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    • pp.57-68
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    • 2018
  • Objectives: To experimentally assess the anti-depressant effects of Gammakdaejo-tang Complex Extracts (GMDJ-Tang) in rats. Methods: Twenty Wistar Hannover (160~170 g) stressed rats were treated with different concentrations of GMDJ-Tang extracts i.e., 0, 62.5 mg/kg, 125 mg/kg, and 250 mg/kg. Chronic mild stress was induced by food deprivation, empty bottles, forced swimming, flickered light, tilt cages, shaking cages, high density breeding, water deprivation, and by soaking the litter cover according to fixed schedule. Blood and brain tissue samples were collected for biochemical analysis. Tests included serotonin and ELISA assays. Results: GMDJ-tang increased the weight of treated rats as well as levels of serotonin, BDNF and TrkB; however, the differences were not significant. In contrast, the extracts significantly decreased blood glucose in stressed rats. GMDJ-tang extracts did not significantly impact Serum AST, ALT, leukocytes, erythrocytes and thrombocytes when comparing treatment groups to control rats. Likewise, hemoglobin, hematocrit and PLT increased in treatment groups following treatment with GMDJ-tang, but this change was without significance. Conclusions: These results suggest that GMDJ-tang can alleviate chronic mild stress in rats, possibly through anti-depressant activity.