• The Korean Journal of Physiology and Pharmacology
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• v.6 no.4
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• pp.183-186
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• 2002

Ischemic damage is one of the most serious problems. The openers of KATP channel have been suggested to have an effect to limit the ischemic damage. However, it is not yet clear how KATP channels of a cell correspond to hypoxic damage. To address the question, N2a cells were exposed to two different hypoxic conditions as follows: 6 hours hypoxia followed by 3 hours reperfusion and 12 hours hypoxia followed by 3 hours reperfusion. As the results, 6 hours hypoxic treatment increased glibenclamide- sensitive basal $K_{ATP}$ current activity (approximately 6.5-fold at 0 mV test potential) when compared with nomoxic condition. In contrast, 12 hours hypoxic treatment induced a relatively smaller change in the $K_{ATP}$ current density (2.5-fold at 0 mV test potential). Additionally, in experiments where $K_{ATP}$ channels were opened using diazoxide, the hypoxia for 6 hours significantly increased the current density in comparison to control condition (p＜0.001). Interestingly, the augmentation in the $K_{ATP}$ current density reduced after exposure to the 12 hours hypoxic condition (p＜0.001). Taken together, these results suggest that $K_{ATP}$ channels appear to be recruited more in cells exposed to the 6 hours hypoxic condition and they may play a protective role against hypoxia-reperfusion damage within the time range.

• Biomedical Science Letters
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• v.17 no.3
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• pp.185-190
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• 2011

Adenovirus type 5 (Ad5) vectors have been used for gene transfer to a wide variety of cell types in vivo and in vitro. The advantages of adenovirus vectors include the high titer of virus readily obtained in large scale preparations, their ability to transduce dividing and non dividing cells, and the high level of transgene expression. Since adenovirus vectors do not integrate in host cell DNA, there is a lack of insertional mutagenesis. However, many human tumor cells lack expression of the adenovirus 5 receptors and contain areas of hypoxia. In order to identify the pattern of replication and gene expression of oncolytic adenovirus in hypoxic condition, multiple different fiber modified Ads (Ad5F/S11, Ad5F/S35, Ad5F/K7, Ad5F/K21, and Ad5F/RGD) was compared. The replication of all fiber modified adenovirus was inhibited in hypoxic condition in HEK 293 cells, but gene expression has variety on different tumor cell lines and the level of coxackievirus and adenovirus receptor (CAR) expression. These data suggest that CAR expression pattern and hypoxic condition of tumor are considered for optimal oncolytic adenovirus application.

• Korean Journal of Exercise Nutrition
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• v.13 no.1
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• pp.45-50
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• 2009

This study evaluated the effects of D-ribose supplementation on exercise time in a hypoxic chamber. Eight healthy young males participated in experiments under all four different conditions; placebo+normoxic, placebo+hypoxic, ribose+normoxic, and ribose+hypoxic. Subjects took 1 g per 10 kg body weight of ribose dissolved in drinking water 30 minutes before and immediately before running. We observed the run-to-exhaustion time, the maximum heart rate, and the changing pattern of the heart rate during exercise. The longest running time was achieved when subjects ran under normoxic condition with ribose supplementation. The shortest running time occurred when subjects ran under hypoxic conditions without ribose supplementation. We measured MDA and GPx to determine any changes in oxidative stresses or antioxidative systems. MDA was affected by the environmental conditions and the running time. The activity of GPx showed a significant difference only with the different environmental conditions of exercise. The results of this study indicate that ribose can be considered a possible ergogenic during exercise in specific conditions, but more detailed and well-controlled studies are needed to make a definitive conclusion.

• The Journal of Pediatrics of Korean Medicine
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• v.19 no.2
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• pp.255-269
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• 2005

Objective : To evaluate the effect of Oriental Medical Treatment on a patient with Hypoxic Ischemic Encephalopathy followed by Cardiopulmonary Resuscitation Method : We applied various methodology of Oriental Medical Treatment including Acupuncture, Electroacupuncture, Physical treatment, Herbal Medicine, Moxibustion treatment and Western medication as well. Result: Herbal medicine was applied on the basis of the patient's history. We applied formular to remove phelgm as a pathogenic factor after Hypoxic Ischemic Encephalopathy. At the same time, considering the patient spent more than a month in ICU lacking appropriate nutrition, we used formuli on the basis of 'Deficiency of Spleen' focusing to vitalize the function of digestive system. As the condition of the patient changed, we also adapted formular accordingly. We prescribed Herbal medication to strengthen Yin and Yang equally as she got hospitalized for long time. Also we applied Acupuncture treatment and Moxibustion treatment to control Qi flow. The general condition of the patient got better with successful removal of Foley catheter and elevated Glasgow Coma scale. We used Electroacupuncture, Physical treatment and Western medication at the same to get maximized effect on relaxing the contracted muscle. According to the Modified Ashworth Scale (MAS), we have some changes in muscle spasticity but later, the effect was not that significant. Conclusion : We had a patient with Hypoxic Ischemic Encephalopathy followed by Cardiopulmonary Resuscitation. In the management of Hypoxic Ischemic Encephalopathy, Conservative treatments are the mainstream but there are not many alternatives. Therefore, We suggest that Oriental medical approach may contribute to the management of Hypoxic Ischemic Encephalopathy.

• Biomaterials Research
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• v.22 no.4
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• pp.271-278
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• 2018

Background: The chondrogenic differentiation of mesenchymal stem cells (MSCs) is regulated by many factors, including oxygen tensions, growth factors, and cytokines. Evidences have suggested that low oxygen tension seems to be an important regulatory factor in the proliferation and chondrogenic differentiation in various MSCs. Recent studies report that synovium-derived mesenchymal stem cells (SDSCs) are a potential source of stem cells for the repair of articular cartilage defects. But, the effect of low oxygen tension on the proliferation and chondrogenic differentiation in SDSCs has not characterized. In this study, we investigated the effects of hypoxia on proliferation and chondrogenesis in SDSCs. Method: SDSCs were isolated from patients with osteoarthritis at total knee replacement. To determine the effect of oxygen tension on proliferation and colony-forming characteristics of SDSCs, A colony-forming unit (CFU) assay and cell counting-based proliferation assay were performed under normoxic (21% oxygen) or hypoxic (5% oxygen). For in vitro chondrogenic differentiation, SDSCs were concentrated to form pellets and subjected to conditions appropriate for chondrogenic differentiation under normoxia and hypoxia, followed by the analysis for the expression of genes and proteins of chondrogenesis. qRT-PCR, histological assay, and glycosoaminoglycan assays were determined to assess chondrogenesis. Results: Low oxygen condition significantly increased proliferation and colony-forming characteristics of SDSCs compared to that of SDSCs under normoxic culture. Similar pellet size and weight were found for chondrogensis period under hypoxia and normoxia condition. The mRNA expression of types II collagen, aggrecan, and the transcription factor SOX9 was increased under hypoxia condition. Histological sections stained with Safranin-O demonstrated that hypoxic conditions had increased proteoglycan synthesis. Immunohistochemistry for types II collagen demonstrated that hypoxic culture of SDSCs increased type II collagen expression. In addition, GAG deposition was significantly higher in hypoxia compared with normoxia at 21 days of differentiation. Conclusion: These findings show that hypoxia condition has an important role in regulating the synthesis ECM matrix by SDSCs as they undergo chondrogenesis. This has important implications for cartilage tissue engineering applications of SDSCs.

• Journal of The Korean Dental Society of Anesthesiology
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• v.14 no.2
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• pp.107-114
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• 2014

Background: Angiogenesis has been recognized an essential precondition for osteogenesis. Because reduction and disruption of the blood supply to tissue cause tissue hypoxia, pathological bone loss affected by hypoxia often can occur in various clinical conditions. The effects of propofol on the process of osteogenesis have received little direct attention. Therefore, we investigated the effect of propofol on the growth and function of osteoblasts under hypoxic condition. Methods: After propofol (3, 30, $300{\mu}M$) preconditioning for 2 hours, hFOB 1.19 human osteoblast cells were cultured under 1 % oxygen tension for 48 hours. Using real time PCR and western blot analysis, we analyzed the expression of, BMP-2, TGF-${\beta}1$, type I collagen, osteocalcin, HIF-1s and Akt. Cell viability was also determined by MTT assay. Results: Propofol preconditioning on hypoxic-cultured osteoblast promoted the expressions of BMP-2, TGF-${\beta}1$, type I collagen and osteocalcin and induced hypoxia-mediated HIF-1 activation and the expression of Akt protein. Propofol with $300{\mu}M$ significant decreased cell viability compared to control. Conclusions: Clinically relevant concentrations of propofol are not cytotoxic to hypoxic osteoblasts in vitro. Propofol preconditioning on hypoxic-cultured osteoblast stimulates proliferation and differentiation of osteoblast through induced expression of BMP-2, TGF-${\beta}1$, type I collagen and osteocalcin. Propofol might promote angiogenesis and bone regeneration under hypoxic condition.

• IMMUNE NETWORK
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• v.2 no.1
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• pp.25-34
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• 2002

Background: Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by synovial hyperplasia and joint destruction. The synovial fibroblasts express cell adhesion molecules and have a role in adhesive interation with inflammatory cells in synovial tissue. It has been suggested that hypoxic conditioins are thought to exist in arthritic joints, and several studies indicate that reactive oxygen species (ROS) produced in hypoxic condition can initiate events that lead to pro-adhesive changes via increased expression of adhesion molecules. So, this study wsa designed to examine whether antioxidant can inhibit hypoxia-induced expression of ICAM-1 in cultured human synovial fibroblasts. Methods: Synovial fibroblasts were isolated from synovial tissue in patients with RA and cultured at hypoxic condition. Antioxidant, PDTC (pyrrolidine dithiocarbamate) were pre-treated for an hour before the hypoxic culture and synovial fibroblasts were harvested at 0, 6, 12, 24, 48 hours time points. Cell surface ICAM-1 expression in synovial fibroblasts was examined by the flow cytometric analysis. To analyse the expression of ICAM-1 mRNA, reverse-transcriptase polymerase chain reaction (RT-PCR) was performed. The levels of cytokines in culture supernatants were measured by ELISA, and activation of NF-${\kappa}B$ was analysed by electrophoretic mobility shift assay. The adhesive reaction between synovial fibroblasts and lymphocytes was assayed by measurement of fluorescent intensity of BCECF-AM in lymphocytes. Results: Hypoxic stimuli up-regulated the ICAM-1 expression as well as the adhesive interaction of human synvial fibroblasts to lymphocytes in a time-dependent manner, and PDTC inhibited hpyoxia-induced ICAM-1 expression and cell-cell interaction. PDTC also inhibited the hypoxia-induced activation of intracellular transcription factor, NF-${\kappa}B$. PDTC decreased the amount of hypoxia-induced production of IL-$1{\beta}$ and TNF-${\alpha}$. Conclusion: These studies demonstrate that PDTC inhibit the hypoxia-induced expression of the adhesion molecule, ICAM-1 and activation of NF-${\kappa}B$ in cultured human synovial fibroblasts.

• Journal of Korean Neurosurgical Society
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• v.29 no.9
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• pp.1161-1170
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• 2000

Objective : The transverse hippocampal slice is one of the most commonly studied in vitro models of mammalian brain physiology. However, despite its broad usage, there has been no standardization of slice preparation techniques or recording condition. It is well known that variations in recording conditions can result in profound different effects to neuronal responses. Evoked field potentials, recorded extracellularly, were used to investigate the effects of variations in hippocampal slice preparation protocol on hypoxia responses of CA1 neurones. Material & Methods : Before hypoxic injury, hippocampal slices were incubated for 4 hours. During incubation period, the slices were placed in a incubation chamber($21^{\circ}C$) for recovery from preparation injury and then transferred to recording chamber($34^{\circ}C$) for more recovery and baseline electric recording with current stimulation(0.1Hz). Various time periods in incubation chamber and recording chamber were applied to each experimental group(group 1=60min : 180min, group 2=90min : 150min, group 3=180min : 60min, time in incubation chamber : time in recording chamber) before 10 min hypoxia produced by replacing 95% $O_2$+5% $CO_2$ mixed gas to 95% $N_2$+5% $CO_2$ gas. Calcium, Magnesium ions and several drugs effecting on glutamate receptor also were studied. Recoveries from hypoxic injury of hippocampal slices were estimated by percent recovery of population spike(PS). Statistic analysis of study were performed using paired t-test. Results : The percent recovery of PS after 10min hypoxia was considerably enhanced by increasing the period of current stimulation during incubation period before hypoxic injury. Temperature effect on the result of this experiment was also studied(group 4) but the result from this showed no statistic significance. Low magnesium ion concentration of artificial CSF(Mg-free aCSF) during incubation period enhanced the recovery of PS but low calcium (calcium-free) and high magnesium ion concentration(2mM) reduced it after hypoxic injury. L-glutamate($100{\mu}M$) and AP-5($50{\mu}M$) had no effect on the recovery of PS but CNQX($10{\mu}M$) in artificial CSF during incubation period markedly enhanced the recovery of PS. Co-treatment of AP-5($50{\mu}M$), CNQX($10{\mu}M$) and high magnesium concentration(2mM) enhanced recovery of PS in immediate following period of hypoxic injury but the effect of cotreatment after then decayed rapidly and lost statistic significance. Conclusions : Judging from above results, the condition of baseline recording is important in observing the recovery of population spike after hypoxia, and the time and the condition should be controled more strictly to obtain reliable results.

• Journal of Korean Neurosurgical Society
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• v.49 no.1
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• pp.13-19
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• 2011

Objective: The purpose of this study is to investigate the combined effects of ginkgo biloba extract, ginkgolide A and B and aspirin on SK-N-MC, human neuroblastoma cell viability and mRNA expression of growth associated protein43 (GAP43), Microtubule-associated protein 2 (MAP2), B-cell lymphoma2 (Bcl2) and protein53 (p53) gene in hypoxia and reperfusion condition. Methods: SK-N-MC cells were cultured with Dulbecco's Modified Eagle's Medium (DMEM) media in $37^{\circ}C$, 5% $CO_2$ incubator. The cells were cultured for 8 hours in non-glucose media and hypoxic condition and for 12 hours in normal media and $O_2$ concentration. Cell survival rate was measured with Cell Counting Kit-8 (CCK-8) reagent assay. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to estimate mRNA levels of GAP43, MAP2, Bcl2, and p53 genes. Results: The ginkgolide A and B increased viable cell number decreased in hypoxic and reperfused condition. The co-treatment of ginkgolide B with aspirin also increased the number of viable cells, however, there was no additive effect. Although there was no increase of mRNA expression of GAP43, MAP2, and Bcl2 in SK-N-MC cells with individual treatment of ginkgolide A, B or aspirin in hypoxic and reperfused condition, the co-treatment of ginkgolide A or B with aspirin significantly increased GAP43 and Bcl2 mRNA levels. In MAP2, only the co-treatment of ginkgolide A and aspirin showed increasing effect. The mRNA expression of p53 had no change in all treating conditions. Conclusion: This study suggests that the combined treatments of Ginkgo biloba extracts and aspirin increase the regeneration of neuroblastoma cells injured by hypoxia and reperfusion.

• Biomolecules & Therapeutics
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• v.15 no.4
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• pp.261-265
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• 2007

Hydroxyproline (HYP) is a post-translational product of proline hydroxylation catalyzed by an enzyme prolyl 4-hydroxylase (P4H) which plays a crucial role in the synthesis of all collagens. Considering the role of collagen and its significance in many clinically important diseases such as liver fibrosis, a great deal of attention has been directed toward the development of an assay at cell-based system. The reason is that cell-based assay system is more efficient than enzyme-based in vitro system and takes much less time than in vivo system. Several assay procedures developed for P4H are laborious, time-consuming and not feasible for the massive-screening. Here, we report the cell-based assay method of prolyl 4-hydroxylase in immortalized rat hepatic stellate HSC-T6 cells. To optimize the cell culture condition to assay for HYP content, various concentrations of reagents were treated for different times in HSC-T6 cells. Our data showed that the treatment with ascorbate in a hypoxic condition for 24 h resulted in the maximal increase of HYP by 1.8 fold. Alternatively, cobalt chloride ($5\;{\mu}M$) and ascorbate ($50\;{\mu}M$) in normoxic states exhibited similar effect on the production of HYP as in a hypoxic condition. Therefore, cobalt chloride can be substituted for a hypoxic condition when an anaerobic chamber is not available. Rosiglitazone and HOE077, known as inhibitors of collagen, synthesis decreased P4H enzyme activity by 32.3% and 15%, respectively, which coincided with previous reports from liver tissues. The level of the smooth muscle ${\alpha}$-actin, a marker of activated stellate cells, was significantly increased under hypoxia, suggesting that our experimental condition could work for screening the anti-fibrotic compounds. The assay procedure took only 3 days after treatment with agents, while assays from the primary stellate cells or liver tissues have taken several weeks. Considering the time and expenses, this assay method could be useful to screen the compounds for the inhibitor of prolyl 4-hydroxylase.