• Archives of Pharmacal Research
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• 제29권1호
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• pp.50-58
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• 2006

Translationally controlled tumor protein (TCTP), also known as histamine releasing factor (HRF), is found abundantly in different eukaryotic cell types. The sequence homology of TCTP between different species is very high, belonging to the MSS4/DSS4 superfamily of proteins. TCTP is involved in both cell growth and human late allergy reaction, as well as having a calcium binding property; however, its primary biological functions remain to be clearly elucidated. In regard to many possible functions, the TCTP of Plasmodium falciparum (Pf) is known to bind with an antimalarial agent, artemisinin, which is activated by heme. It is assumed that the endoperoxide-bridge of artemisinin is opened up by heme to form a free radical, which then eventually alkylates, probably to the Cys14 of PfTCTP. Study of the docking of artemisinin with heme, and subsequently with PfTCTP, was carried out to verify the above hypothesis on the basis of structural interactions. The three dimensional (3D) structure of PfTCTP was built by homology modeling, using the NMR structure of the TCTP of Schizosaccharomyces pombe as a template. The quality of the model was examined based on its secondary structure and biological function, as well as with the use of structure evaluating programs. The interactions between artemisinin, heme and PfTCTP were then studied using the docking program, FlexiDock. The center of the peroxide bond of artemisinin and the Fe of heme were docked within a short distance of $2.6{\AA}$, implying the strong possibility of an interaction between the two molecules, as proposed. When the activated form of artemisinin was docked on the PfTCTP, the C4-radical of the drug faced towards the sulfur of Cys14 within a distance of $2.48{\AA}$, again suggesting the possibility of alkylation having occurred. These results confirm the proposed mechanism of the antimalarial effect of artemisinin, which will provide a reliable method for establishing the mechanism of its biological activity using a molecular modeling study.

• The Korean Journal of Physiology and Pharmacology
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• 제26권3호
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• pp.165-174
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• 2022

As the mechanism underlying glucose metabolism regulation by oxymatrine is unclear, this study investigated the effects of oxymatrine on pyroptosis in INS-1 cells. Flow cytometry was employed to examine cell pyroptosis and reactive oxygen species (ROS) production. Cell pyroptosis was also investigated via transmission electron microscopy and lactate dehydrogenase (LDH) release. Protein levels were detected using western blotting and interleukin (IL)-1β and IL-18 secretion by enzyme-linked immunosorbent assay. The caspase-1 activity and DNA-binding activity of nuclear factor kappa B (NF-κB) and nuclear factor (erythroid-derived 2)-like 2 protein (Nrf2) were also assessed. In the high glucose and high fat-treated INS-1 cells (HG + PA), the caspase-1 activity and LDH content, as well as Nod-like receptor family pyrin domain containing 3, Gsdmd-N, caspase-1, apoptosis-associated speck-like protein containing a CARD, IL-1β, and IL-18 levels were increased. Moreover, P65 protein levels increased in the nucleus but decreased in the cytoplasm. Oxymatrine attenuated these effects and suppressed high glucose and high fat-induced ROS production. The increased levels of nuclear Nrf2 and heme oxygenase-1 (HO-1) in the HG + PA cells were further elevated after oxymatrine treatment, whereas cytoplasmic Nrf2 and Keleh-like ECH-associated protein levels decreased. Additionally, the elevated transcriptional activity of p65 in HG + PA cells was reduced by oxymatrine, whereas that of Nrf2 increased. The results indicate that the inhibition of pyroptosis in INS-1 cells by oxymatrine, a key factor in its glucose metabolism regulation, involves the suppression of the NF-κB pathway and activation of the Nrf2/HO-1 pathway.

• 대한의생명과학회지
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• 제11권3호
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• pp.349-353
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• 2005

The globin in cytosol receives the heme in mitochondria and folds to the hemoglobin within erythrocyte. Two mechanisms have been proposed that the heme was transfered post-translationally or cotranslationally to the globin. In this research, how the globin in cytosol receives post-translationally the heme from membranes was studied according to pH and phospholipid composition. Globins dissolved in various pH buffer solutions $(PH\;3\~7)$ were rapidly added into the bulk of egg phosphatidylcholine $100\%\;or\;60\%$ liposomes containing hemin in pH 7 buffer solution. Hemin was very highly transferred to globin at pH 4 and 6. Also, hemin was more efficiently transfered to globin in egg phosphatidylcholine $100\%$ than in $60\%$ liposomes.

• Food Quality and Culture
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• 제1권1호
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• pp.46-57
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• 2007

The purpose of the present study was to examine the nutrient intake status of elderly dementia patients. We surveyed the dietary intake of 50 demented elderly patients receiving a regular diet, who were hospitalized in a geriatric hospital in Yongin, Gyeonggi-do. The average age of the subjects was $79.6{\pm}6.5$ for the males and $80.5{\pm}6.3$ for the females. The average heights were $169.5{\pm}6.0\;cm$ (males) and $154.6{\pm}5.2\;cm$ (females), and the average weights were $58.5{\pm}7.4$ (males) and $51.7{\pm}8.9\;kg$ (females). Depending on the type of dementia, the male patients showed significant differences in their intakes of vegetable protein, fiber, total iron, and non-heme iron between the Alzheimer's disease (AD) and the vascular dementia (VD) groups, and the female patients showed significant differences in their intakes of total protein, phosphorus, zinc, and vitamin $B_6$ between the two groups. The male patients showed significant differences in their intakes of animal-source calcium and carotene according to ability to self-feed. According to physical activity, the male patients showed significant differences in vegetable fat and vitamin A intake, and the females showed significant differences in their intakes of total protein, animal protein, fiber, ash, total calcium, vegetable-source calcium, animal-source calcium, phosphorous, total iron, non-heme iron, heme iron, sodium, vitamins $B_1$, $B_2$, $B_6$, niacin, vitamin C, folate, and potassium. We found that the patients consumed excessive protein, but low amounts of calcium, vitamin $B_2$, and folate. Also, the patients' physical activity abilities appeared to affect their nutrient intakes.

• Parasites, Hosts and Diseases
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• 제48권1호
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• pp.15-21
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• 2010

Astrocytes are the most abundant cells in the central nervous system that play roles in maintaining the blood-brain-barrier and in neural injury, including cerebral malaria, a severe complication of Plasmodium falciparum infection. Prostaglandin (PG) $D_2$ is abundantly produced in the brain and regulates the sleep response. Moreover, $PGD_2$ is a potential factor derived from P. falciparum within erythrocytes. Heme oxygenase-1 (HO-1) is catalyzing enzyme in heme breakdown process to release iron, carbon monoxide, and biliverdin/bilirubin, and may influence iron supply to the P. falciparum parasites. Here, we showed that treatment of a human astrocyte cell line, CCF-STTG1, with $PGD_2$ significantly increased the expression levels of HO-1 mRNA by RT-PCR. Western blot analysis showed that $PGD_2$ treatment increased the level of HO-1 protein, in a dose- and time-dependent manner. Thus, $PGD_2$ may be involved in the pathogenesis of cerebral malaria by inducing HO-1 expression in malaria patients.

• Nutrition Research and Practice
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• 제8권4호
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• pp.352-359
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• 2014

BACKGROUND/OBJECTIVES: In this study, we determined the anti-inflammatory activities and the underlying molecular mechanisms of the methanol extract from Erigeron Canadensis L. (ECM) in LPS-stimulated RAW264.7 macrophage cells. MATERIALS/METHODS: The potential anti-inflammatory properties of ECM were investigated by using RAW264.7 macrophages. We used western blot assays and real time quantitative polymerase chain reaction to detect protein and mRNA expression, respectively. Luciferase assays were performed to determine the transactivity of transcription factors. RESULTS: ECM significantly inhibited inducible nitric oxide synthase (iNOS)-derived NO and cyclooxygenase-2 (COX-2) derived PGE2 production in LPS-stimulated RAW264.7 macrophages. These inhibitory effects of ECM were accompanied by decreases in LPS-induced nuclear translocations and transactivities of $NF{\kappa}B$. Moreover, phosphorylation of mitogen-activated protein kinase (MAPKs) including extracellular signal-related kinase (ERK1/2), p38, and c-jun N-terminal kinase (JNK) was significantly suppressed by ECM in LPS-stimulated RAW264.7 macrophages. Further studies demonstrated that ECM by itself induced heme oxygenase-1 (HO-1) protein expression at the protein levels in dose-dependent manner. However, zinc protoporphyrin (ZnPP), a selective HO-1 inhibitor, abolished the ECM-induced suppression of NO production. CONCLUSIONS: These results suggested that ECM-induced HO-1 expression was partly responsible for the resulting anti-inflammatory effects. These findings suggest that ECM exerts anti-inflammatory actions and help to elucidate the mechanisms underlying the potential therapeutic values of Erigeron Canadensis L.

• IMMUNE NETWORK
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• 제20권3호
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• pp.26.1-26.9
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• 2020

Cereblon (CRBN), a negative modulator of AMP-activated protein kinase (AMPK), is highly expressed in the retina. We confirmed the expression of CRBN in ARPE-19 human retinal cells by Western blotting. We also demonstrated that CRBN knock-down (KD) could effectively downregulate IL-6 and MCP-1 protein and gene expression in LPS-stimulated ARPE-19 cells. Additionally, CRBN KD increased the phosphorylation of AMPK/acetylcoenzyme A carboxylase (ACC) and the expression of heme oxygenase-1 (HO-1) in ARPE-19 cells. Furthermore, CRBN KD significantly reduced LPS-induced nuclear translocation of NF-κB p65 and activation of NF-κB promoter activity. However, these processes could be inactivated by compound C (inhibitor of AMPK) and zinc protoporphyrin-1 (ZnPP-1; inhibitor of HO-1). In conclusion, compound C and ZnPP-1 can rescue LPS-induced levels of proinflammatory cytokines (IL-6 and MCP-1) in CRBN KD ARPE-19 cells. Our data demonstrate that CRBN deficiency negatively regulates proinflammatory cytokines via the activation of AMPK/HO-1 in the retina.

• Bulletin of the Korean Chemical Society
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• 제23권6호
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• pp.865-872
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• 2002

A systematic way of determining the equilibrium orientation of carbon monoxide (CO) in heme proteins using time-resolved polarized mid-IR spectroscopy is presented. The polarization anisotropy at pump-probe delay time of zero in the limit of zero photolysis and the angular distrbution function of CO are required to obtain the equilibrium orientation of CO. An approach is developed for determining the polarization anisotropy in the zero-photolysis limit from the anisotropy measured under finite photolysis conditions in an optically thick sample where the fraction of molecules photolyzed decreased as the pump pulse passes through and is absorbed by the sample. This approach is verified by measuring the polarization anisotropy of CO of carbonmonoxy myoglobin at various levels of photolysis. This method can be readily applied to other photoselection experiments determining precise angle between transition dipoles.

• Journal of Photoscience
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• 제9권3호
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• pp.33-36
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• 2002

This study describes RT-PCR and in situ hybridisation protocols, and the immunohistochemical detection method that we have developed to detect and localise cells that express HO-1 in the skin. We found that HO-1 mRNA was absent in normal mouse skin, but after UVA irradiation HO-1 mRNA was expressed in the dermal fibroblasts, and strongly in basal epidermal cells. HO-1 protein was also induced strongly in dermal fibroblasts, and also in epidermal cells. In addition, the HO substrate heme was reduced in skin microsome at 72 hrs post UVA (when HO activity is high). At the same time, the HO products bilirubin and iron levels were elevated in the cutaneous tissue. Thus in addition to a dermal response, there appears to be an epidermal HO response to UVA in vivo that may be relevant for immune modulation by UVA radiation.

• Bulletin of the Korean Chemical Society
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• 제20권5호
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• pp.543-546
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• 1999

Heme assignment of the 1HNMR spectrum of cytochrome c3 of D. vulgaris Miyazaki F was established [Reference: 12, 13]. The major reduction of the heme turned out to take place in the other of heme 4, 1, 2 and 3 (in the sequential numbering). The Hemes with the smallest and greatest solvent accessibility were reduced at the highest and lowest potentials in average, respectively. A cooperation interheme interaction was attributed to a pait of the closest hemes, namely, hemes 1 and 2. This assignment can provide the physicochemical bases for the elucidation of electron transfer of this protein.