• Biomolecules & Therapeutics
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• v.16 no.3
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• pp.190-196
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• 2008

Deoxypodophyllotoxin (DPT) is a medicinal herb product isolated from Anthriscus sylvestris. DPT possesses beneficial activities in regulating immediate-type allergic reaction and anti-inflammatory activity through the dual inhibition of cyclooxygenase-2 and 5-lipoxygenase. In the present study, the metabolism of DPT was further characterized in rat liver microsomes isolated from male Sprague Dawley rats. The metabolism of DPT was NADPH-dependent. In addition, when liver microsomes were incubated with SKF-525A, a well-known CYP inhibitor, in the presence of $\beta$-NADPH, the metabolism of DPT was significantly inhibited. Using enriched rat liver microsomes, the anticipated isoforms of cytochrome P450s (CYPs) in the metabolism of DPT were partially characterized. Phenobarbital-induced microsomes increased in the formation of metabolite M1. The metabolite M3 was only produced in the enriched microsomes isolated from dexamethasone-treated rats. The results indicated that the metabolism of DPT would be CYP-dependent and that CYP2B and CYP3A might be important in the metabolism of DPT in rats.

• The Korean Journal of Physiology and Pharmacology
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• v.19 no.6
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• pp.491-497
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• 2015

Traumatic brain injury (TBI) is a major cause of mortality and long-term disability, which can decrease quality of life. In spite of numerous studies suggesting that Epigallocatechin-3- gallate (EGCG) has been used as a therapeutic agent for a broad range of disorders, the effect of EGCG on TBI remains unknown. In this study, a weight drop model was established to evaluate the therapeutic potential of EGCG on TBI. Rats were administered with 100 mg/kg EGCG or PBS intraperitoneally. At different times following trauma, rats were sacrificed for analysis. It was found that EGCG (100 mg/kg, i.p.) treatment significantly reduced brain water content and vascular permeability at 12, 24, 48, 72 hour after TBI. Real-time PCR results revealed that EGCG inhibited TBI-induced IL-$1{\beta}$ and TNF-${\alpha}$ mRNA expression. Importantly, CD68 mRNA expression decreasing in the brain suggested that EGCG inhibited microglia activation. Western blotting and immunohistochemistry results showed that administering of EGCG significantly inhibited the levels of aquaporin-4 (AQP4) and glial fibrillary acidic protein (GFAP) expression. TBI-induced oxidative stress was remarkably impaired by EGCG treatment, which elevated the activities of SOD and GSH-PX. Conversely, EGCG significantly reduced the contents of MDA after TBI. In addition, EGCG decreased TBI-induced NADPH oxidase activation through inhibition of $p47^{phox}$ translocation from cytoplasm to plasma membrane. These data demonstrate that EGCG treatment may be an effective therapeutic strategy for TBI and the underlying mechanism involves inhibition of oxidative stress.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.5
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• pp.295-302
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• 2002

To understand the utilization property of light energy, Synechococcus sp. MA19, a poly-${\beta}$-hydroxybutyrate (PHB) producer, was cultivated at the different incident light intensities of 15.3, 50.0 and 78.2 W/$m^2$ using media with and without phosphate. From the results of metabolic flux analysis, it was found that the cell yield based on ATP synthesis was estimated as $3.5{\times}10^{-3}$ kg-biomass/mol-ATP in these cultures. Under the examined conditions, there were no significant differences in the efficiency of light energy conversion to chemical energies estimated as ATP synthesis and reducing potential (NADH + NADPH) formation whether the PHB synthesis took place or not. The energy converted from light to ATP was kept relatively high around the energy absorbed by the cells of $2.5-3.0{\times}10^{6} J\;h^{-1}\;kg^{-1}$, whereas the energy of reducing potential was hardly changed in the examined range of the energy absorbed by the cells.

• Journal of Microbiology and Biotechnology
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• v.9 no.2
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• pp.140-146
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• 1999

When methanol was the sole carbon source, Methylobacterium organophilum NCIB 11278, a facultative methylotroph, accumulated Poly-$\beta$-hydroxybutyrate (PHB) as 59% (w/w) of dry cell weight under potassium limitation, 37% under sulfate limitation, and 33% under nitrogen limitation. Based on a stoichiometric analysis of PHB synthesis from methanol, it was suspected that PHB synthesis is accompanied by the overproduction of energy, either 6-10 ATP and 1 $FADH_2$ or 6 ATP and 3 NADPH to balance the NADH requirement, per PHB monomer. This was confirmed by observation of increased intracellular ATP levels during PHB accumulation. The intracellular ATP with limited potassium, sulfate, and ammonium increased to 0.185, 0.452, and 0.390 $\mu$moles ATP/g Xr (residual cell mass) during PHB accumulation, respectively. The intracellular ATP level under potassium limitation was similar to that when there was no nutrient limitation and no PHB accumulation, 0.152- 0.186 $\mu$moles ATP/g Xr. We propose that the maximum PHB accumulation observed when potassium was limited is a result of the energy balance during PHB accumulation. Microorganisms have high energy requirements under potassium limitation. Enhanced PHB accumulation, in ammonium and sulfate limited conditions with the addition of 2,4-dinitrophenol, which dissipates surplus energy, proves this assumption. With the addition of 1 mM of 2,4-dinitrophenol, the PHB content increased from 32.4% to 58.5% of dry cell weight when nitrogen limited and from 15.1 % to 31.0% of dry cell weight when sulfate limited.

• Journal of Life Science
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• v.8 no.6
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• pp.695-701
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• 1998

This study was carried out to investigate levels of the components of microsomal mixed function oxidase (MFO) system and activities of the hepatic microsomal cytochrome P45O (P45O)-dependent monooxygenases of grass snake (Natrix tigrina Lateralis) and to compare with those of rat. The levels of P45O and cytochrome b$_{5}$, (b$_{5}$) of snake were much lower than those in rat. NADPH-cytochrome c reductase activity in the snake was also only 40% of that in the rat. Activities of 7-ethoxycoumarin 0-deethylase (ECOD) and benzphetamine N-demethylase (BPDM) of snake hepatic microsomes, when compared with those of rat, were markedly low. But, aryl hydrocarbon hydroxylase (AHH) and testosterone hydroxylase (TSH) activities were nearly the same or higher than those of the rat. Of the P45O-dependent TSHs measured, 7$\alpha$-hydroxylase activity was the highest in snake, whereas, 6$\beta$-hydroxylase activity was the highest in rat. However, stereoselectivity of the enzyme from the snake to C2 and C6 positions of testoste-rone was the same as rat. The result of radioimmunoassay (RIA) for the identification of five P45O isozymes with MAbs shows that relatively high content of ethanol-inducible P45O isozyme, CYP2El, exists in the rat, whereas MC-inducible P45O isozyme, CYP2A1/1A2, does in the snake. From the analyses of SDS-PAGE and RIA of partially pu-rified P45O, we suggest the possibility of the presence of a certain P45O isozyme(s) in hepatic microsomes of snake different from those of rat.

• Journal of Ginseng Research
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• v.45 no.6
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• pp.665-675
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• 2021

Background: Ginsenoside Rg1 (Rg1), an active ingredient in ginseng, may be a potential agent for the treatment of Alzheimer's disease (AD). However, the protective effect of Rg1 on neurodegeneration in AD and its mechanism of action are still incompletely understood. Methods: Wild type (WT) and APP/PS1 AD mice, from 6 to 9 months old, were used in the experiment. The open field test (OFT) and Morris water maze (MWM) were used to detect behavioral changes. Neuronal damage was assessed by hematoxylin and eosin (H&E) and Nissl staining. Immunofluorescence, western blotting, and quantitative real-time polymerase chain reaction (q-PCR) were used to examine postsynaptic density 95 (PSD95) expression, amyloid beta (Aβ) deposition, Tau and phosphorylated Tau (p-Tau) expression, reactive oxygen species (ROS) production, and NAPDH oxidase 2 (NOX2) expression. Results: Rg1 treatment for 12 weeks significantly ameliorated cognitive impairments and neuronal damage and decreased the p-Tau level, amyloid precursor protein (APP) expression, and Aβ generation in APP/PS1 mice. Meanwhile, Rg1 treatment significantly decreased the ROS level and NOX2 expression in the hippocampus and cortex of APP/PS1 mice. Conclusions: Rg1 alleviates cognitive impairments, neuronal damage, and reduce Aβ deposition by inhibiting NOX2 activation in APP/PS1 mice.

• The Korean Journal of Physiology and Pharmacology
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• v.15 no.6
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• pp.319-326
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• 2011

Quercetin-3-O-${\beta}$-D-glucuronopyranoside (QGC) is a flavonoid glucoside extracted from Rumex Aquaticus Herba. We aimed to explore its protective effect against ethanol-induced cell damage and the mechanism involved in the effect in feline esophageal epithelial cells (EEC). Cell viability was tested and 2',7'-dichlorofluorescin diacetate assay was used to detect intracellular $H_2O_2$ production. Western blotting analysis was performed to investigate MAPK activation and interleukin 6 (IL-6) expression. Exposure of cells to 10% ethanol time-dependently decreased cell viability. Notably, exposure to ethanol for 30 min decreased cell viability to 43.4%. When cells were incubated with $50{\mu}M$ QGC for 12 h prior to and during ethanol treatment, cell viability was increased to 65%. QGC also inhibited the $H_2O_2$ production and activation of ERK 1/2 induced by ethanol. Pretreatment of cells with the NADPH oxidase inhibitor, diphenylene iodonium, also inhibited the ethanol-induced ERK 1/2 activation. Treatment of cells with ethanol for 30 or 60 min in the absence or presence of QGC exhibited no changes in the IL-6 expression or release compared to control. Taken together, the data indicate that the cytoprotective effect of QGC against ethanol-induced cell damage may involve inhibition of ROS generation and downstream activation of the ERK 1/2 in feline EEC.

• The Journal of Korean Medicine
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• v.22 no.3
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• pp.92-97
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• 2001

목적 :본 연구는 최근 임상에서 많이 사용하는 자하거 약침액이 과산화수소($H_2O_2$)로 야기된 송과선 세포의 Apoptosis에 있어서 세포 보호에 미치는 영향과 그 기전을 분석하였다. 방법 :송과선 세포주에서 자하거 약침액의 $H_2O_2$로 인한 Apoptosis에 대한 방어 기전을 관찰하기 위하여 면역세포화학법. 세포화학법 및 reverse transcription-polymerase chain reaction (RT-PCR)을 시행하였다. 결과 : 자하거 약침액 투여군에서는 nuclear factor kappa B (NFkB), inducible nitric oxide synthase (iNOS), nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase의 발현이 $H_2O_2$ 투여군보다 감소하였다. RT-PCR에서는 caspase-3의 발현이 자하거 약침액 투여군에서 $H_2O_2$ 투여군보다 억제되었다. 결론： 이상의 결과를 통하여 자하거 약침액이 $H_2O_2$로 유발된 Apoptosis에서 세포보호 효과가 있으며 그 기전은 iNOS와 caspase-3의 억제에 기인할 가능성을 시사한다고 하겠다.

• Journal of Korean Medicine Rehabilitation
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• v.23 no.3
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• pp.55-68
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• 2013

Objectives This study was to investigate the effects of Yuhyangjeongtong-san on the carrageenin-induced acute inflammation and adjuvant-induced arthritis in rats. Methods Acute inflammation was induced by injection of 2% carrageenin 0.15 ml into right hind foot of rats. Control group was carrageenin injected and taken distilled water, treated group was carrageenin injected and taken Yuhyangjeongtong-san by orally once. 2 hours after injection, plantar temperature and paw volume were measured. 3 hours after, counts of white blood cell (WBC) were performed. Arthritis was induced by injection of complete freund's adjuvant (CFA) into base of tail. Control group was CFA injected and taken distilled water, treated group was CFA injected and taken Yuhyangjeongtong-san by orally for 10 days. 0,5,10 day, body weight, thickness of ankle joint and paw edema were measured. 10 day, counts of WBC, $interleukin-1{\beta}$ ($IL-1{\beta}$) and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) level were analysed. Histochemical study of NADPH-d and immunohistochemical study of c-fos were performed. Results In the carrageenin-induced acute inflammation, neutrophils of treated group were significantly decreased compared with control group. In the adjuvant-induced arthritis, paw edema, total counts of WBC and TNF-${\alpha}$ level in paw exudates were significantly decreased compared with control group, and the number of c-fos positive neurons of treated group was significantly decreased compared with control group. Conclusions On the basis of these results, we concluded that Yuhyangjeongtong-san have anti-arthritic and anti-inflammatory effects.

• Biomolecules & Therapeutics
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• v.31 no.5
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• pp.515-525
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• 2023

The most common heart valve disorder is calcific aortic valve stenosis (CAVS), which is characterized by a narrowing of the aortic valve. Treatment with the drug molecule, in addition to surgical and transcatheter valve replacement, is the primary focus of researchers in this field. The purpose of this study is to determine whether niclosamide can reduce calcification in aortic valve interstitial cells (VICs). To induce calcification, cells were treated with a pro-calcifying medium (PCM). Different concentrations of niclosamide were added to the PCM-treated cells, and the level of calcification, mRNA, and protein expression of calcification markers was measured. Niclosamide inhibited aortic valve calcification as observed from reduced alizarin red s staining in niclosamide treated VICs and also decreased the mRNA and protein expressions of calcification-specific markers: runt-related transcription factor 2 and osteopontin. Niclosamide also reduced the formation of reactive oxygen species, NADPH oxidase activity and the expression of Nox2 and p22^phox. Furthermore, in calcified VICs, niclosamide inhibited the expression of β-catenin and phosphorylated glycogen synthase kinase (GSK-3β), as well as the phosphorylation of AKT and ERK. Taken together, our findings suggest that niclosamide may alleviate PCM-induced calcification, at least in part, by targeting oxidative stress mediated GSK-3β/β-catenin signaling pathway via inhibiting activation of AKT and ERK, and may be a potential treatment for CAVS.