• Archives of Pharmacal Research
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• v.20 no.2
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• pp.103-109
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• 1997

The effects of ginseng total saponins (GTS) on hypoxic damage of primary cultures of astrocytes were studied. Hypoxia was created by placing cultures in an air tight chamber that was flushed with 95% $N_2/5%CO_2$ for 15 min before being sealed. Cultures showed evidence of significant cell injury after 24 h of hypoxia (increased lactate dehydrogenase (LDH) content in the culture medium, cell swelling and decreased glutamate uptake and protein content). Addition of GTS (0.1, 0.3 mg/ml) to the cultures during the exposure to hypoxic conditions produced dose-dependent inhibition of the LDH efflux. GTS (0.1, 0.3 mg/ml) also produced significant inhibition of the increased cell volume of astrocytes measured by $[^3H]$ O-methyl-D-glucose uptake under the hypoxic conditions. Decreased glutamate uptake and protein content was inhibited by GTS. These data suggest that GTS prevents astrocytic cell injury induced by severe hypoxia in vitro.

• Korean Journal of Exercise Nutrition
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• v.24 no.3
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• pp.7-12
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• 2020

[Purpose] This study was performed to investigate the acid-base and ion balance at rest and after exercise in healthy males under normoxia, moderate hypoxia, and severe hypoxia. [Methods] Ten healthy Korean males completed three different trials on different days, comprising exercise under normoxia (F_iO₂ = 20.9%, N trial), moderate hypoxia (F_iO₂ = 16.5%, MH trial), and severe hypoxia (F_iO₂ = 12.8%, SH trial). They undertook endurance exercise for 30 min on a cycle ergometer at the same relative exercise intensity equivalent to 80% maximal heart rate under all conditions. Capillary blood samples were obtained to determine acid-base and ion balance at rest and after exercise. [Results] Exercise-induced blood lactate elevations were significantly increased as hypoxic conditions became more severe; SH > MH > N trials (P = 0.003). After exercise, blood glucose levels were significantly higher in the SH trial than in the N and MH trials (P = 0.001). Capillary oxygen saturation (S_CO₂) levels were significantly lowered as hypoxic conditions became more severe; SH > MH > N trials (P < 0.001). The pH levels were significantly lower in the MH trial than that in the N trial (P = 0.010). Moreover, H_CO_3^- levels were significantly lower in the SH trial than in the N trial, with significant interaction (P = 0.003). There were no significant differences in blood Na⁺, K⁺, and Ca²⁺ levels between the trials. [Conclusion] MH and SH trials induced greater differences in glucose, lactate, S_CO₂, pH, and HCO_3^- levels in capillary blood compared to the N trial. Additionally, lactate, S_CO₂, and HCO_3^- levels showed greater changes in the SH trial than in the MH trial. However, there were no significant differences in Na⁺, K⁺, and Ca²⁺ levels in MH and SH trials compared to the N trial.

• Journal of The Korean Dental Society of Anesthesiology
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• v.14 no.1
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• pp.49-56
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• 2014

Background: Propofol (2.6-diisopropylphenol) is a widely used intravenous anesthetic agent for the induction and maintenance of anesthesia during surgeries and sedation for ICU patients. Propofol has a structural similarity to the endogenous antioxidant vitamin E and exhibits antioxidant activities.13) However, the mechanism of propofol on hypoxia/reoxygenation (H/R) injury has yet to be fully elucidated. We investigated how P-PostC influences the autophagy and cell death, a cellular damage occurring during the H/R injury. Methods: The groups were randomly divided into the following groups: Control: cells were incubated in normoxia (5% CO2, 21% O2, and 74% N2) without propofol treatment. H/R: cells were exposed to 24 h of hypoxia (5% CO2, 1% O2, and 94% N2) followed by 12 h of reoxygenation (5% CO2, 21% O2, and 74% N2). H/R + P-PostC: cells post-treated with propofol were exposed to 24 h of hypoxia followed by 12 h of reoxygenation. 3-MA + P-PostC: cells pretreated with 3-MA and post-treated propofol were exposed to 24 h of hypoxia followed by 12 h of reoxygenation Results: The results of our present study provides a new direction of research on mechanisms of propofol-mediated cytoprotection. There are three principal findings of these studies. First, the application of P-PostC at the onset of reoxygenation after hypoxia significantly increased COS-7 cell viability. Second, the cellular protective effect of P-PostC in H/R induced COS-7 cells was probably related to activation of intra-cellular autophagy. And third, the autophagy pathway inhibitor 3-MA blocked the protective effect of P-PostC on cell viability, suggesting a key role of autophagy in cellular protective effect of P-PostC. Conclusions: These data provided evidence that P-PostC reduced cell death in H/R model of COS-7 cells, which was in agreement with the protection by P-PostC demonstrated in isolated COS-7 cells exposed to H/R injury. Although the this study could not represent the protection by P-PostC in vivo, the data demonstrate another model in which endogenous mechanisms evoked by P-PostC protected the COS-7 cells exposed to H/R injury from cell death.

• International Journal of Oral Biology
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• v.39 no.2
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• pp.97-105
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• 2014

The aim of this study was to determine the beneficial effect of propofol on human keratinocytes that have undergone hypoxia reoxygenation (H/R) injury and to investigate whether autophagy is associated with the protective mechanism. Thus, we evaluated how propofol influences the intracellular autophagy and apoptosis during the H/R process in the HaCaT cells. The cultured human keratinocyte cells were exposed to 24 h of hypoxia (5% $CO_2$, 1% $O_2$, 94% $N_2$) followed by 12 h of reoxygenation (5% $CO_2$, 21% $O_2$, 74% $N_2$). The experiment was divided into 4 groups: (1) Control=Normoxia ; (2) H/R=Hypoxia Reoxygenation ; (3) PPC+H/R=Propofol Preconditioning+Hypoxia Reoxygenation; (4) 3-MA+PPC+ H/R=3-MA-Methyladenine+Propofol Preconditioning+ Hypoxia Reoxygenation. In addition, Western blot analysis was performed to identify the expression of apoptotic pathway parameters, including Bcl-2, Bax, and caspase 3 involved in mitochondrial-dependent pathway. Autophagy was determined by fluorescence microscopy, MDC staining, AO staining, and western blot. The H/R produced dramatic injuries in keratinocyte cells. In our study, the viability of Propofol in H/R induced HaCaT cells was first studied by MTT assay. The treatment with 25, 50, and $100{\mu}M$ Propofol in H/R induced HaCaT cells enhanced cell viability in a dose-dependent manner and $100{\mu}M$ was the most effective dose. The Atg5, Becline-1, LC3-II, and p62 were elevated in PPC group cells, but H/R-induced group showed significant reduction in HaCaT cells. The Atg5 were increased when autophagy was induced by Propofol, and they were decreased when autophagy was suppressed by 3-MA. These data provided evidence that propofol preconditioning induced autophagy and reduced apoptotic cell death in an H/R model of HaCaT cells, which was in agreement with autophagy playing a very important role in cell protection.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1998.11a
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• pp.138-138
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• 1998

Effects of nitric oxide and hypoxia on ethoxyresorufin deethylase in Hepa I cells and MCF-7 human breast cancer cells were examined. TCDD treatment have resulted in the stimulation of ethoxyresorufin deethylase activity based on fluorometry in Hepa I in dose and time dependent manner. 0.1 nM TCDD showed maximal stimulation of ethoxyresorufin deethylase activity and 24 hour treatment also showed maximal stimulation of ethoxyresorufin deethylase activity. In MCF-7 human breast cancer cells, untreated cells showed high basal level of ethoxyresorufin deethylase activity. TCDD treatment to MCF-7 cells resulted minor stimulation of ethoxyresorufin deethylase activity compared to that in Hepa I cells. Nitric oxide and hypoxia inhibit TCDD effects on ethoxyresorufin deethylase activity in both cell lines. And also flavonoids, such as quercetin showed an inhibition of ethoxyresorufin deethylase activity that is stimulated with TCDD or 3-Methylcholanthrene. Estrogen and estrogen metabolite such as 16 a-estriol and 2-hydroxyestradiol also affects the ethoxyresorufin deethylase activity in MCF-7 cells.

• Journal of Yeungnam Medical Science
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• v.15 no.1
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• pp.97-113
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• 1998

The sympathoadrenal system plays an important role in homeostasis in widely varing external environments. Conflicting findings, however, have been reported on its response to hypoxia. We investigated the effect of hypoxia on the sympathoadrenal system in dogs under halothane anesthesia by measuring levels of circulating catecholamines in response to graded hypoxia. Ten healthy mongreal dogs were mechanically ventilated with different hypoxic gas mixtures. Graded hypoxia and reoxygenation were induced by progressively decreasing the oxygen fraction in the inhalation gas mixture from 21%(control) to 15%, 10% and 5% at every 5 minutes, and then reoxygenated with 60% oxygen. Mean arterial pressure, central venous pressure and mean pulmonary arterial pressure were measured directly using pressure transducers. Cardiac output was measured by the thermodilutional method. For analysis of blood gas, saturation and content, arterial and mixed venous blood were sampled via the femoral and pulmonary artery at the end of each hypoxic condition. The concentration of plasma catecholamines was determined by radioenzymatic assay. According to the exposure of graded hypoxia, not only did arterial and mixed venous oxygen tension decreased markedly at 10% and 5% oxygen, but also arterial and mixed venous oxygen saturation decreased significantly. An increased trend of the oxygen extraction ratio was seen during graded hypoxia. Cardiac output, mean arterial pressure and systemic vascular resistance were unchanged or increased slightly. Pulmonary arterial pressure(PAP) and pulmonary vascular resistance(PVR) were increased by 55%, 76% in 10% oxygen and by 82%, 95% in 5% oxygen, respectively(p<0.01). The concentrations of plasma norepinephrine, epinephrine and dopamine increased by 75%, 29%, 24% in 15% oxygen and by 382%, 350%, 49% in 5% oxygen. These data suggest that the sympathetic nervous system was activated to maintain homeostasis by modifying blood flow distribution to improve oxygen delivery to tissues by hypoxia, but hemodynamic changes might be blunted by high concentration of nitrous oxide except PAP and PVR. It would be suggested that hemodynamic changes might not be sensitive index during hypoxia induced by high concentration of nitrous oxide exposure.

• Clinical and Experimental Reproductive Medicine
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• v.37 no.3
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• pp.217-229
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• 2010

Objective: Apoptosis plays an important role for the maintenance of the normal pregnancy. Expression of X-linked inhibitor of apoptosis (XIAP) is able to effectively prevent apoptosis and controls trophoblast cells death throughout placental development, but it is still unknown in the function of XIAP in trophoblast cells exposed to hypoxic condition, which is one of the factors causing preeclampsia. Therefore, we conducted to compare XIAP expression in normal and pre-eclamptic placenta tissues and analyzed the function of XIAP in HTR-8/SVneo trophoblast cell line exposed to hypoxic condition. Methods: The expression of XIAP was analyzed in placental tissues from the following groups of patients (none underwent labor): 1) term normal placenta (n=15); 2) term with pre-eclamptic placeneta (n=15); and 3) pre-term with pre-eclamptic placenta (n=11) using semi-quantitative RT-PCR, immunohistochemistry, and Western blot. In order to evaluate the function of XIAP in HTR-8/SVneo trophoblast cells under hypoxic condition, HIF-$1{\alpha}$ plasmids, and hypoxic condtion were transfected and treated into HTR-8/SVneo trophoblast cells for 24 hours, respectively. Results: We observed that XIAP are expressed in the syncytiotrophoblasts and syncytial knot of placental villi. The expression of XIAP was significantly decreased in preeclamptic placenta tissues than in normal placenta tissues without labor (p<0.05). Furthermore, we confirmed the XIAP expression in HTR-8/SVneo trophbolast cells exposed to hypoxia was translocated from cytoplasm into nucleus and decreased XIAP by hypoxic condition induced apoptosis in HTR-8/SVneo trophoblast cells through up-regulation of pro-apoptotic proteins. Conclusion: These results suggest that the expression of XIAP is involved in placental development as well as decreased expression of XIAP by hypoxia is associated with pre-eclampsia through inducing trophoblast cells apoptosis.

• Journal of The Korean Dental Society of Anesthesiology
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• v.14 no.2
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• pp.101-106
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• 2014

Background: Remifentanil, an ultra-short-acting mu-opioid receptor agonist, is unique from other opioids because of its esterase-based metabolism, minimal accumulation, and very rapid onset and offset of clinical action. Remifentanil can prevent the inflammatory response and can suppress inducible nitric oxide synthase expression in a septic mouse model. However, the effects of remifentanil on human keratinocyte and autophagy have yet to be fully elucidated during hypoxia-reoxygenation. Here we investigated whether remifentanil confers protective effect against hypoxia-reoxygenation in human keratinocyte and, if so, whether autophagy mediates this effect. Methods: The human keratinocytes were cultured under 1% oxygen tension. The cells were gassed with 94% $N_2$, and 5% $CO_2$ and incubated for 24 h at $37^{\circ}C$. To determine whether the administration of affects human keratinocytes hypoxia-reoxygenation injury, cells were then exposed to various concentrations of remifentanil (0.01, 0.1, 0.5 and 1 ng/ml) for 2 h. After remifentanil treatment, to simulate reoxygenation and recovery, the cells were reoxygenated for 12 h at $37^{\circ}C$. Control group did not receive remifentanil treatment. Normoxia group did not receive hypoxia and remifentanil treatment for 36 h. 3-MA group was treated 3-methyladenine (3-MA) for 1h before remifentanil treatment. Cell viability was measured using a quantitative colorimetric assay with MTT, showing the mitochondrial activity of living cells. Cells were stained with fluorescence and analyzed with Western blot analysis to find out any relations with activation of autophagy. Results: Prominent accumulation of autophagic specific staining MDC was observed around the nuclei in RPT group HaCaT cells. Similarly, AO staining, red fluorescent spots appeared in RPT group HaCaT cells, while the Normoxia, control and 3-MA groups showed mainly green cytoplasmic fluorescence. We here examined activation of autophagy related protein under H/R-induced cells by Western blotting analysis. Atg5, Beclin-1, LC3-II (microtubule-associated protein 1 light chain 3 form II) and p62 was elevated in RPT group cells. But they were decreased when autophagy was suppressed by 3-MA (Fig. 5). Conclusions: Although the findings of this study are limited to an in vitro interpretation, we suggest that remifentanil may have a beneficial effect in the recovery of wound from hypoxia-reoxygenation injury.

• Journal of Korean Medicine Rehabilitation
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• v.24 no.2
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• pp.41-50
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• 2014

Objectives The purpose of this study was to evaluate the effects of Clematidis radix extract on $CoCl_2$-induced apoptosis in SH-SY5Y human neuroblastoma cells. Methods In order to investigate the protective effect of Clematidis radix on $CoCl_2$-induced cytotoxicity in neuronal cells, MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, DAPI(4,6-diamidino-2-phenylindoleI) staining, TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling) assay, DNA fragmentation assay and western blotting were performed on SH-SY5Y human neuroblastoma cells. Results Cells treated with $CoCl_2$ exhibited several apoptotic features, while cells pre-treated with Clematidis radix prior to $CoCl_2$ exposure showed a decrease in the occurrence of apoptotic features. $CoCl_2$ increased HIF-$1{\alpha}$ expression, in contrast, Clematidis radix treatment decreased $CoCl_2$-induced HIF-$1{\alpha}$ expression. Pre-treatment with the extract of Clematidis radix suppressed Bax, cytochrome c, and caspase-3 expressions, and also increased Bcl-2 expression in SH-SY5Y human neuroblastoma cells. Conclusions These results suggest that Clematidis radix may exert a protective effect on $CoCl_2$-induced apoptosis in SH-SY5Y human neuroblastoma cells.

• Clinical and Experimental Reproductive Medicine
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• v.36 no.3
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• pp.163-174
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• 2009

Objectives: Members of the immortalization-upregulated protein (IMUP) family are nuclear proteins implicated in SV40-mediated immortalization and cellular proliferation, but the mechanisms by which their expression is regulated are still unknown in placenta. To investigate to expression and functions of IMUPs in placenta, we conducted to compare IMUPs expression in normal and preeclamptic placenta tissues and analyzed the function of IMUP-2 in HTR-8/SVneo trophoblast cells after IMUP-2 gene transfection. Methods: The expression of IMUPs was analyzed in placental tissues from the following groups of patients (none underwent labor): 1) term normal placenta (n=15); 2) term with preeclamptic placeneta (n=15); and 3) pre-term with preeclamptic placenta (n=11) using semi-quantitative RT-PCR, RNA in situ hybiridization, immunohistochemistry, and Western blot. In order to evaluate the function of IMUP-2 in HTR-8/SVneo trophoblast cells, IMUP-2 plasmids were transfected into HTR-8/SVneo trophoblast cells for 24 hours. Results: We observed that IMUPs are mainly expressed in the syncytiotrophoblasts and syncytial knot of placental villi. The expression of IMUP-1 was not differences between normal and preeclamptic placenta tissues. However, IMUP-2 expression was significantly higher in preterm preeclamptic placenta tissues than in normal placenta tissues without labor (p<0.001). Furthermore, we confirmed overexpression of IMUP-2 induced apoptosis in HTR-8/SVneo trophoblast cells through up-regulation of pro-apoptotic proteins. Conclusions: These results suggest that the expression of IMUP-2 is involved in placental development as well as increased IMUP-2 expression is associated with preeclampsia through the inducing of trophoblast apoptosis.