• Journal of Applied Biological Chemistry
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• 제65권3호
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• pp.167-172
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• 2022

Pathophysiological reaction of platelets in the blood vessel is an indispensable part of thrombosis and cardiovascular disease, which is the most common cause of death in the world. In this study, we performed in vitro assays to evaluate antiplatelet activity of artemether in human platelets and attempted to identify the mechanism responsible for protein phosphorylation. Artemether is a derivative of artemisinin, known as an active ingredient of Artemisia annua, which has been reported to be effective in treating malaria, and is known to function through antioxidant and metabolic enzyme inhibition. However, the role of artemether in platelet activation and aggregation and the mechanism of action of artemether in collagen-induced human platelets are not known until now. In this study, the effect of artesunate on collagen-induced human platelet aggregation was confirmed and the mechanism of action of artemether was clarified. Artemether inhibited the phosphorylation of PI3K/Akt and Mitogen-activated protein kinases, which are phosphoproteins that are known to act in the signal transduction process when platelets are activated. In addition, artemether decreased TXA₂ production and decreased granule secretion in platelets such as ATP and serotonin release. As a result, artemether strongly inhibited platelet aggregation induced by collagen, a strong aggregation inducer secreted from vascular endothelial cells, with an IC₅₀ of 157.92 μM. These results suggest that artemether has value as an effective antithrombotic agent for inhibiting the activation and aggregation of human platelets through vascular injury.

• Applied Microscopy
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• 제28권3호
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• pp.363-377
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• 1998

After the investigation on the eye of Incilaria fruhstorieri with light and electron microscopes, the following results were obtained. The eye of Incilaria fruhstorferi comprises cornea, lens, vitreous body, retina, and optic nerve inward from the outside. Cornea is composed of squamous, cuboid, columnar and irregular cells, which appear to be light due to their low electron density. In their cytoplasms, glycogen granules, multivesicular body, and nucleus were observed. Vitreous body, located behind non-cellular transparent lens, is filled with long and short microvilli protruding from the retinal epithelia. Retinal epithelium, the organ to perceive objects, is divided into four parts; microvillar layer pigment layer, nuclear layer, and neutrophils layer, from the apical portion. Microvillar layer consists of the type-I photoreceptor cells and pigmented granule cells. In the apical portion of their cytoplasms, long microvilli (length, $19{\mu}m$) , short microvilli (length, $8{\mu}m$), and rolled microvilli grow thick in the irregular and mixed forms. Photoreceptor cells are classified into type-I and type-II, according to their structures. The type-I cell has the apical portion rising roundly like a fan and the lower part which looks like the helve of a fan. In the cytoplasm of the apical portion, there are clear vesicles, cored vesicles, ovoid mitochondria, and microfilaments, and in the cytoplasm of the lower part, photic vesicles with their diameters about 60nm aggregate densely. The type-II photoreceptor cell, located at the lower end of the type-I cells, has a very large ovoid nucleus 3nd no microvilli. In the cytoplasm of the type-II cell, the photic vesicles with sizes 60nm aggregate more densely than in the cytoplasm of the type-I cell. Pigmented cells are classified into type-A and type-B, according to their structures. The type-A is identified to be a large cell containing round granules (diameter, $0.5{\mu}m$) of very high electron density, while the type-B is identified as a small cell where the irregular granules (diameter, $0.6{\mu}m$) of a little lower electron density amalgamate. Nuclear layer ranges from the bottom of pigment layer to the top of the capsule, and contains three kinds of nuclei (nuclei of the type-II photoreceptor cell, pigmented granule cell, and accessory neuron). The capsules covering the outmost part of the eyeball are composed of collagenous fiber and three longitudinal muscle layers (the thickness of each longitudinal muscle layer, $0.4{\mu}m$) and thick circular muscle layer (thickness, $0.3{\mu}m$). Around the capsules, there is a neurophile layer consisting of neurons and nerve fibers. Each neuron has a relatively large ovoid nucleus for its cytoplasm, and in the karyosome, large lumps of keterochromatin form a wheel nucleus.

• The Korean Journal of Physiology and Pharmacology
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• 제9권1호
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• pp.9-15
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• 2005

We attempted to analyze the mechanism of polychlorinated biphenyl (PCB)-induced neurotoxicity and identify the target molecules in the neuronal cells for PCBs.Since the developing neuron is particularly sensitive to PCB-induced neurotoxicity, we isolated cerebellar granule cells derived from 7-day old Sprague Dawley (SD) rats and grew cells in culture for additional 7 days to mimic PND-14 conditions. Only non-coplanar PCBs at a high dose showed a significant increase of total protein kinase C (PKC) activity at phobol 12,13-dibutyrate ([$^3M$]PDBu) binding assay, indicating that non-coplanar PCBs are more neuroactive than coplanar PCBs in neuronal cells. PKC isozymes were immunoblotted with the selected monoclonal antibodies. PKC-${\alpha}$, ${\delta}$, and ε were activated with non-coplanar PCB exposure. Receptor for activated C kinase-1 (RACK-1), anchoring protein for activated PKC, was more induced with exposure to coplanar PCBs than non-coplanar PCBs. Reverse transcription PCR (RT-PCR) analysis showed induction of neurogranin (RC-3) and growth associated protein-43 (GAP-43) mRNA with non-coplanar PCBs. The results indicate that these factors may be useful biomarkers for differentiating non-coplanar PCBs from coplanar PCBs. The present study demonstrated that non-coplanar PCBs are more neuroactive congeners than coplanar PCBs.

• Applied Microscopy
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• 제36권2호
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• pp.109-118
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• 2006

자외선 B를 사람의 두피에서 채취한 모발에 6시간, 12시간, 24시간. 48시간 동안 각각 조사한 후 모발의 미세구조적인 변화 과정과 모발 단백질의 분해에 의한 원소 성분의 함량 변화를 전자현미경 및 에너지분산분광분석기를 이용하여 분석하였다. 자외선에 6시간 조사된 모발에서 모발 표면은 중첩된 큐티클층이 분리되어 있거나 부러져있는 상태로 나타났다. 큐티클세포의 내큐티클에는 직경이 30nm에서부터 700nm크기의 공포를 형성하고 있었다. 자외선에 12시간 조사된 모발에서 피질은 치밀하게 배열되어 있는 각화세포의 세포막이 파괴되어 공포가 형성되었는데 공포는 세포막을 따가서 일렬로 배열되어 있었다. 또한, 멜라닌 과립과 접촉하고 있는 거대원섬유에서 공포가 형성되어 멜라닌 과립과 분리되는 것을 확인할 수 있었다. 이 시기에도 멜라닌 과립의 모양이나 크기의 변화는 나타나지 않았다. 자외선에 24시간 조사된 모발에서 큐티클세포들은 절단된 상태로 관찰되었고 세포질의 내큐티클에 형성된 공포들은 인접해 형성된 공포들과 융합되었다. 자외선에 48시간 조사된 모발은 표면이 거칠게 나타났고, 큐티클의 안쪽 층에 있는 세포에서부터 형성된 공로로 인해 위쪽에 있는 큐티클 세포는 들뜨면서 조각나 떨어져 나갔다. 자외선 조사를 받아 손상된 모발의 원소성분을 에너지 분산 분광분석기로 분석한 결과 정상모발 보다 산소성분의 함량이 높게 나타났으나 황의 성분은 감소되었다.

• Molecules and Cells
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• 제41권3호
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• pp.214-223
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• 2018

Oligoadenylate synthetase (OAS) protein family is the major interferon (IFN)-stimulated genes responsible for the activation of RNase L pathway upon viral infection. OAS-like (OASL) is also required for inhibition of viral growth in human cells, but the loss of one of its mouse homolog, OASL1, causes a severe defect in termination of type I interferon production. To further investigate the antiviral activity of OASL1, we examined its subcellular localization and regulatory roles in IFN production in the early and late stages of viral infection. We found OASL1, but not OASL2, formed stress granules trapping viral RNAs and promoted efficient RLR signaling in early stages of infection. Stress granule formation was dependent on RNA binding activity of OASL1. But in the late stages of infection, OASL1 interacted with IRF7 transcripts to inhibit translation resulting in down regulation of IFN production. These results implicate that OASL1 plays context dependent functions in the antiviral response for the clearance and resolution of viral infections.

• Natural Product Sciences
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• 제9권4호
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• pp.249-255
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• 2003

Zizypus is one of the herbs widely used in Korea and China due to CNS calming effect. The present study aims to investigate the effect of the methanol extract of Zizyphi Jujube Semen (ZJS) on kainic acid (KA)-induced neurotoxicity in cultured rat cerebellar granule neuron. ZJS, over a concentration range of 0.05 to $5\;{\mu]g/ml$, inhibited KA $(500\;{\mu}M)-induced$ neuronal cell death, which was measured by a trypan blue exclusion test and a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay. Pretreatment of ZJS $(0.5\;{\mu}g/ml)$ inhibited KA$(50\;{\mu}M)$-induced elevation of cytosolic calcium concentration $([Ca^{2+}]_c)$, which was measured by a fluorescent dye, Fura 2-AM, and generation of reactive oxygen species (ROS). ZJS $(0.5\;{\mu}g/ml)$ inhibited glutamate release into medium induced by KA $(500\;{\mu}M)$, which was measured by HPLC. These results suggest that ZJS prevents KA-induced neuronal cell damage in vitro.

• Biomolecules & Therapeutics
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• 제16권4호
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• pp.328-335
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• 2008

This study was undertaken to investigate whether honokiol could enhance the pentobarbitalinduced sleeping behaviors through $\gamma$-aminobutyric acid (GABA) receptor $Cl^-$ channel activation. Thirty minutes after the oral administration of honokiol, mice were received sodium pentobarbital (42 mg/kg, i.p.). The time elapsed from pentobarbital injection to the loss of the righting reflex was taken as sleeping latency. The time elapsed between the loss and voluntary recovery of the righting reflex was considered as the total sleeping time. Western blot technique and $Cl^-$ sensitive fluorescence probe were used to detect the expression of $GABA_A$ receptor subunits and $Cl^-$ influx in the primary cultured cerebellar granule cells. Honokiol (0.1 and 0.2 mg/kg) prolonged the sleeping time induced by pentobarbital (42 mg/kg) in a dosage-dependent manner. Honokiol (20 and 50 ${\mu}M$) increased $Cl^-$ influx in primary cultured cerebellar granule cells, and selectively increased the $GABA_A$ receptor $\alpha$-subunit expression, but had no effect on the abundance of $\beta$ or $\gamma$-subunits. Chronic treatment with 20 ${\mu}M$ honokiol in primary cultured cerebellar neurons did not affect the abundance of GAD65/67. The results suggested that honokiol could potentiate pentobarbital-induced sleeping through $GABA_A$ receptor $Cl^-$ channel activation.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2008년도 Proceedings of the Convention
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• pp.119-142
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• 2008

Zizyphi Spinosi Semen (ZSS) has been widely used for the treatment of anxiety and insomnia in Korea and China. This experiment was performed to know whether sanjoinine A, one of major alkaloid compounds of ZSS has anxiolytic and hypnotic effects through the GABAergic systems. Our results showed that administration of sanjoinine A increased open arm entries and spent time in open arm in the elevated plus-maze and increased head dips in hole board test. Different from traditional anxiolytic, diazepam, sanjoinine A itself did not decrease locomotor activity and strength level in mice. Furthermore, Sanjoinine A (0.5-2.0 mg/kg) prolonged sleeping time and reduced sleeping latency induced by pentobarbital in a dose-dependent manner similar to muscimol, a $GABA_A$ receptor agonist. Sanjoinine A (0.25-1.0 mg/kg) also increased sleeping rate and sleeping time in the combined administration at the sub-hypnotic dose of pentobarbital and showed synergic effects with muscimol in potentiating sleeping onset and enhancing sleeping time induced by pentobarbital. However, sanjoinine A itself did not induce sleeping at the higher dose. In addition, both of sanjoinine A and pentobarbital increased chloride influx in primary cultured cerebellar granule cells. Sanjoinine A decreased the $GABA_A$ receptor ${\alpha}$-subunit expression and increased ${\gamma}$-subunit expression, and had no effects on abundance of ${\beta}$-subunit in primary cultured cerebellar granule cells, showing different expression of subunits from pentobarbital. In conclusion, sanjoinine A shows anxiolytic-like effects and augments pentabarbital-induced sleeping behaviors through the modification of GABAergic systems. [This work was supported by the Korea Research Foundation Grant funded by the Korean Government (MOEHRD) (The Regional Research Universities Program/Center for Healthcare Technology Development)].

• 한국약용작물학회지
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• 제11권4호
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• pp.298-305
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• 2003

Polygalae Radix (PR) from Polygala tenuifolia. (Polygalaceae) is traditionally used in China and Korea, since this herb has a sedative, antiinflammatory, and antibacterial agent. To extend pharmacological actions of PR in the CNS on the basis of its CNS inhibitory effect, the present study examined whether PR has the neuroprotective action against kainic acid (KA) -induced cell death in primarily cultured rat cerebellar granule neurons. PR, over a concentration range of 0.05 to $5{\mu}g/ml$ inhibited KA $(500\;{\mu}M)$-induced neuronal cell death, which was measured by a trypan blue exclusion test and a 3-[4,5-dimethylthiazol-2-y1]-2,5-diphenyl-tetrazolium bromide (MTT) assay. PR $(0.5{\mu}g/ml)$ inhibited glutamate release into medium induced by KA $(500\;{\mu}M)$, which was measured by HPLC. Pretreatment of PR $(0.5{\mu}g/ml)$ inhibited KA $(500\;{\mu}M)$-induced elevation of cytosolic calcium concentration $([Ca^{2+}]_c)$ which was measured by a fluorescent dye, Fura 2-AM, and generation of reactive oxygen species (ROS). These results suggest that PR prevents KA-induced neuronal cell damage in vitro.

• IMMUNE NETWORK
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• 제6권4호
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• pp.179-184
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• 2006

Background: Identification of antigen-specific T cells has yielded valuable information on pathologic process and the disease state. Assays for quantification of inflammatory cytokines or lytic-granule molecules have been generally used to evaluate antigen specific T cell response, however their applicability have been hampered due to the limited source of autologous antigen-presenting target cells (APC). Methods: K562, a leukemic cell line deficient of human leukocyte antigen (HLA), was transfected with a gene encoding HLA-A*02 (K562/ A*02) and its function as stimulator cells in inducing activation of HLA-matched T cells was evaluated by IFN-${\gamma}$ enzyme linked immunospot (ELISPOT) assay. Results: The stable transfectant K562/ A*02 pulsed with HLA- A*02 restricted peptide could specifically induce IFN-${\gamma}$ secretion by CD8+ T cells compared to no detectable secretion by CD4+ T cells. However, CD56+ NK cells secreted IFN-${\gamma}$ in both K562/ A*02 with peptide and without peptide. The number of IFN-${\gamma}$ secreted CD8+ T cells was increased according to the ratio of T cells to K562 and peptide concentration. Formalin-fixed K562/ A*02 showed similar antigen presenting function to live K562/ A*02. Moreover, K562/ A*02 could present antigenicpeptide to not only A*0201 restricted CD8+ T cells but also CD8+ T cells from A*0206 donor. Conclusion: These results suggest that K562/ A*02 could be generally used as target having specificity and negligible background for measuring CD8+ T cell responses and selective use of K562 with responsder matched HLA molecules on its surface as APC may circumvent the limitation of providing HLA-matched autologous target cells.