• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.2
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• pp.244-249
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• 2003

To investigate the hepatic oxygen free radical metabolizing system and changes of serum cholesterol levels in rats fed a diet supplemented with Monascus pigment (MP), Sprague-Dawley rats weighing about 300 g have been fed a diet supplemented with 2% or 4% MP for a month. The rats fed 2% MP supplemented diet gained less body weight than the control rats and those fed 4% W supplemented diet. Those fed 2% or 4% MP supplemented diet had no remarkable changes in liver function on basis of liver weight/body weight, serum levels of xanthine oxidase, alanine amino transferase activity In rats fed 2% and 4% MP supplemented diet, hepatic cytochrome P45O dependent aniline hydroxylase activity significantly (p<0.05) declined about 32%, 37% respectively and showed no significant differences between rats fed 2% and 4% MP supplemented diet whereas those fed 2% MP supplemented diet showed about 29% increased hepatic xanthine oxidase activity. And hepatic glutathione S-transferase and glutathione peroxidase activites in rats fed 2% MP supplemented were more increased by about 17%, 28% respectively than the control rats. There were no significant differences both in between those fed 2% and 4% MP supplemented diet. Especially rats fed 2% or 4% MP supplemented diet showed a significant (p<0.05) increase in hepatic catalase activity by 41%, 25% compared with control rats and those fed 4% MP supplemented diet showed more decrease in tendency of catalase activity than those 2% MP supplemented diet. But hepatic superoxide dismutase activity and glutathione content were appeared to be similar value among three groups. On the other hand, rats fed 2% MP supplement diet showed 17% increased levels of serum HDL-choresterol and 26% decreased value of LDL-cholesterol and serum level of triglyceride. But no different value were appeared between those fed 2% and 4% MP supplemented diet. Especially in those fed 2% and 4% MP supplemented diet, artherogenic index were significantly (p<0.05) declined by 37%, 29% respectively compared with control. In conclusion, it is likely that rats fed a diet supplemented with a proper quantity of MP may have the potential of oxygen free radical detoxication and lowering of artherogenic index.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.04a
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• pp.205-205
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• 1996

본 실험은 간장허혈 및 재관류시 야기되는 간장 손상에 대해 vitamin C와 E 각각의 효과와 이들의 병용효과를 알아보고자 하였다. 실험군은 흰쥐에 vitamin E(25mg/kg)를 실험전 3일간 투여한 군, vitamin C(100mg/kg)를 실험 5분전 경정맥주사한 군 및 vitamin C와 E의 병용 투여군등의 3군으로 하여 각각에 허혈을 유발시킨 후 (60분) 재관류 1시간, 5시간에 간세포 손상정도(AI.T, AST, liver wet-weight to dry-weight ratio), 지질과산화(MDA), 담즙분비변동(bile flow, bilirubin, cholate output) 및 약물대사효소계의 변동(cytochrome P$_{450}$, aminopyrine-N-demethylase, aniline p-hydroxylase activity) 등을 관찰하였다. 실험결과로는 허혈 및 재관류로 인한 ALT, AST MDA는 재관류 5시간에 최고치를 이루었으며 이는 vitamin C와 vitamin E의 각각 투여로 억제되었고, 특히 vitamin C와 E의 병용투여로 더욱 현저하게 억제되었다. 간세포 부종의 지표인 liver wet-weight to dry-weight ratio도 vitamin C와 E의 병용투어로 유의성있게 억제되었다. 담즙분비량 및 담즙산량은 vitamin C 투여와 vitamin C와 E 병용투여로 허혈 및 재관류로 감소된 양을 증가시켰고, 특히 vitamin C와 E의 병용투여는 담즙분비량에 있어 현저한 상승을 나타내었다. 허혈 및 재관류로 인한 cytochrome P$_{450}$양의 감소와 aminopyrine N-demethylase 활성의 억제는 vitamin C 투여와 vitamin C와 E의 병용투여에 의해 유의성 있게 증가하였다. 이상의 결과로 보아 vitamin C와 vitamin E는 각각 허혈 및 재관류로 인한 간장손상을 완화시켰으며 특히 vitamin C와 E의 병용투여는 상승적으로 적용하여 간세포손상을 더욱 억제시킴을 알 수 있었다.

• Nutrition Research and Practice
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• v.10 no.5
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• pp.501-506
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• 2016

BACKGROUNG/OBJECTIVES: Corn silk (CS) extract contains large amounts of maysin, which is a major flavonoid in CS. However, studies regarding the effect of CS extract on cholesterol metabolism is limited. Therefore, the purpose of this study was to determine the effect of CS extract on cholesterol metabolism in C57BL/6J mouse fed high-fat diets. MATERIALS/METHODS: Normal-fat group fed 7% fat diet, high-fat (HF) group fed 25% fat diet, and high-fat with corn silk (HFCS) group were orally administered CS extract (100 mg/kg body weight) daily. Serum and hepatic levels of total lipids, triglycerides, and total cholesterol as well as serum free fatty acid, glucose, and insulin levels were determined. The mRNA expression levels of acyl-CoA: cholesterol acyltransferase (ACAT), cholesterol 7-alpha hydroxylase (CYP7A1), farnesoid X receptor (FXR), lecithin cholesterol acyltransferase (LCAT), low-density lipoprotein receptor, 3-hyroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA reductase), adiponectin, leptin, and tumor necrosis factor ${\alpha}$ were determined. RESULTS: Oral administration of CS extract with HF improved serum glucose and insulin levels as well as attenuated HF-induced fatty liver. CS extracts significantly elevated mRNA expression levels of adipocytokines and reduced mRNA expression levels of HMG-CoA reductase, ACAT, and FXR. The mRNA expression levels of CYP7A1 and LCAT between the HF group and HFCS group were not statistically different. CONCLUSIONS: CS extract supplementation with a high-fat diet improves levels of adipocytokine secretion and glucose homeostasis. CS extract is also effective in decreasing the regulatory pool of hepatic cholesterol, in line with decreased blood and hepatic levels of cholesterol though modulation of mRNA expression levels of HMG-CoA reductase, ACAT, and FXR.

• Journal of Microbiology and Biotechnology
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• v.25 no.6
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• pp.782-787
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• 2015

In this study, we developed an assay system for missense mutations in human phenylalanine hydroxylases (hPAHs). To demonstrate the reliability of the system, eight mutant proteins (F39L, K42I, L48S, I65T, R252Q, L255V, S349L, and R408W) were expressed in a mutant strain (pah^-) of Dictyostelium discoideum Ax2 disrupted in the indigenous gene encoding PAH. The transformed pah - cells grown in FM minimal medium were measured for growth rate and PAH activity to reveal a positive correlation between them. The protein level of hPAH was also determined by western blotting to show the impact of each mutation on protein stability and catalytic activity. The result was highly compatible with the previous ones obtained from other expression systems, suggesting that Dictyostelium is a dependable alternative to other expression systems. Furthermore, we found that both the protein level and activity of S349L and R408W, which were impaired severely in protein stability, were rescued in HL5 nutrient medium. Although the responsible component(s) remains unidentified, this unexpected finding showed an important advantage of our expression system for studying unstable proteins. As an economic and stable cell-based expression system, our development will contribute to mass-screening of pharmacological chaperones for missense PAH mutations as well as to the in-depth characterization of individual mutations.

• Journal of Microbiology and Biotechnology
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• v.25 no.10
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• pp.1634-1639
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• 2015

Cytochrome P450 hydroxylase (CYP) in actinomycetes plays an important role in the biosynthesis and bioconversion of various secondary metabolites. Two unique CYPs named CYP-sb21 and CYP-pa1, which were identified from Sebekia benihana and Pseudonocardia autotrophica, respectively, were proven to transfer a hydroxyl group at the 4^th or 9^th N-methyl leucine position of immunosuppressive agent cyclosporin A (CsA). Interestingly, these two homologous CYPs showed different CsA regio-selectivities. CYP-sb21 exhibited preferential hydroxylation activity at the 4^th position over the 9^th position, whereas CYP-pa1 showed the opposite preference. To narrow down the CYP domain critical for CsA regio-selectivity, each CYP was divided into four domains, and each domain was swapped with its counterpart from the other CYP. A total of 18 hybrid CYPs were then individually tested for CsA regio-selectivity. Although most of the hybrid CYPs failed to exhibit a significant change in regio-selectivity in the context of CsA hydroxylation, hybrid CYP-pa1 swapped with the second domain of CYP-sb21 showed a higher preference for the 9^th position. Moreover, hybrid CYPsb21 containing seven amino acids from the 2nd domain of CYP-pa1 showed higher preference for the 4^th position. These results imply that the 2nd domain of CsA-specific CYP plays a critical role in CsA regio-selectivity, thereby setting the stage for biotechnological application of CsA regio-selective hydroxylation.

• Journal of Microbiology and Biotechnology
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• v.17 no.4
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• pp.655-662
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• 2007

This study examined the effects of Lactobacillus acidophilus ATCC 43121 (LAB) on cholesterol metabolism in hypercholesterolemia-induced rats. Four treatment groups of rats (n=9) were fed experimental diets: normal diet, normal $diet+LAB(2{\times}10^6\;CFU/day)$, hypercholesterol diet (0.5% cholesterol, w/w), and hypercholesterol diet+LAB. Body weight, feed intake, and feed efficiency did not differ among the four groups. Supplementation with LAB reduced total serum cholesterol (25%) and VLDL+IDL+LDL cholesterol (42%) in hypercholesterol diet groups, although hepatic tissue cholesterol and lipid contents were not changed. In the normal diet group, cholesterol synthesis (HMG-CoA reductase expression), absorption (LDL receptor expression), and excretion via bile acids (cholesterol $7{\alpha}-hydroxylase$ expression) were increased by supplementation with LAB, and increased cholesterol absorption and decreased excretion were found in the hypercholesterol diet group. Total fecal acid sterols excretion was increased by supplementation with LAB. With proportional changes in both normal and hypercholesterol diet groups, primary bile acids (cholic and chenodeoxycholic acids) were reduced, and secondary bile acids (deoxycholic and lithocholic acids) were increased. Fecal neutral sterol excretion was not changed by LAB. In this experiment, the increase in insoluble bile acid (lithocholic acid) reduced blood cholesterol level in rats fed hypercholesterol diets supplemented with LAB. Thus, in the rat, L. acidophilus ATCC 43121 is more likely to affect deconjugation and dehydroxylation during cholesterol metabolism than the assimilation of cholesterol into cell membranes.

• Korean Journal of Pharmacognosy
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• v.46 no.1
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• pp.84-92
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• 2015

Tyrosinase is a key enzyme to control the biosynthesis of melanin pigments and has two enzyme activities, namely of 1-tyrosine hydroxylase and of 1-dopa oxidase. Thus, tyrosinase is regarded as a target in skin-whitening and therapeutic intervention of local hyperpigmentation diseases. We have tested tyrosinase inhibitory activity on the water extracts of 50 species oriental medicinal plant. Among them, five medicinal plants, Linderae Radix, Clematidis Radix, Cinnamomi Cortex Spissus, Fritillariae Thunbergii Bulbus and Bulbus Fritillariae Cirrhosae were investigated strong inhibition effect. Five medicinal plants were fractionated using organic solvents (methylene chloride, ethyl acetate, n-butanol, water). Cinnamomi Cortex Spissus (ethyl acetate fraction) was investigated strong inhibition effect. Tyrosinase inhibitory activity below $IC_{50}\;40{\mu}g/ml$ is confirmed in five herbal plants that are Linderae Radix, Clematidis Radix, Cinnamomi Cortex Spissus, Fritillariae Thunbergii Bulbus and Bulbus Fritillariae Cirrhosae. Tyrosinase inhibitory levels ($IC_{50}\;{\mu}g/ml$) of each plants were 15.56, 35.02, 25.14, 15.20 and 39.77. We also investigate the effect of effective plant's fraction. in dose of $100{\mu}g/ml$, Cinnamomi Cortex Spissus (P-36) EtOAc fraction significant inhibitory effect over 50%. Clematidis Radix (P-35) and Cinnamomi Cortex Spissus (P-36) MC fraction inhibit tyrosinase each 36.60% and 43.21%. inhibitory rates of Fritillariae Thunbergii Bulbus (P-40) EtOAc and $H_2O$ fraction are 31.40% and 31.51%. Bulbus Fritillariae Cirrhosae (P-45) BuOH fraction regulate tyrosinase activity to 37.71%. We examined tyrosinase inhibitory activity of natural products and these results suggest that several herbs have potential as a new whitening material.

• Animal cells and systems
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• v.10 no.4
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• pp.203-209
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• 2006

Azoles are widely used antifungal agents, which inhibit the biosynthesis of fungal cell-membrane ergosterol. In this study, using an amphibian follicle culture system, the effects of azoles on follicular steroidogenesis in frogs were examined. Itraconazole (ICZ), clotrimazole (CTZ) and ketoconazole (KCZ) suppressed pregnenolone ($P_5$) production by the follicles ($ED_{50};\;0.04_{\mu}M,\;0.33_{\mu} M,\;and\;0.91_{\mu}M$, respectively) in response to frog pituitary homogenates (FPH). However, fluconazole (FCZ), miconazole (MCZ) and econazole (ECZ) were not effective in the suppression of $P_5$ production. Not all the azoles examined suppressed the conversion of exogenous $P_5$ to progesterone ($P_4$) (by $3{\beta}$- HSD) or $P_4$ to $17{\alpha}$-hydroxyprogesterone ($17{\alpha}$-OHP) (by $17{\alpha}$-hydroxylase), or androstenedione (AD) to testosterone (T) (by $17{\beta}$-HSD). In contrast, CTZ, MCZ and ECZ in medium partially suppressed the conversion of $17{\alpha}$-OHP to AD (by C17-20 lyase) ($ED_{50};\;0.25{\mu} M,\;4.5{\mu}M,\;and\;0.7{mu}M$, respectively) and CTZ, KCZ, ECZ and MCZ strongly suppressed the conversion of exogenous T to estradiol ($E_2$) (by aromatase) ($ED_{50};\;0.02{\mu}M,\;8{\mu}M,\;0.07{\mu}M,\;0.8{\mu}M$, respectively). These results demonstrated that some azole agents strongly suppress amphibian follicular steroidogenesis and particularly, P450scc and aromatase are more sensitive to azoles than other steroidogenic enzymes.

• Biomedical Science Letters
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• v.9 no.2
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• pp.75-84
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• 2003

To evaluate the influence of hepatic oxygen free radical systems on liver injury by topical p-phenylenediamine (PPD) application on rat skin, PPD (25 mg/16.5 $\textrm{cm}^2$) was topically applied to the abdominal region 5 times every other day and sacrificed. By PPD treatment, increasing rate of liver weight/body weight (%), serum activities of alanine aminotransferase and aspartate aminotransferase and decreasing rate of microsomal glucose-6-phosphatase activity were higher in the rats fed tungstate supplemented diet than those fed a standard diet. These findings indicate that group fed tungstate supplemented diet have more severe liver injury compared with group fed standard diet on topical PPD application. However, the activities of oxygen free radical generating enzymes such as xanthine oxidase (XO) and cytochrome P450 dependent aniline hydroxylase and those of oxygen free radical scavenging enzymes were not found to be different between these two animal groups. In the present study, a novel monitoring method to detect the generating of oxygen free radicals in liver extract was devised. Throughout this method, the oxidized PPD produced by oxygen free radicals was determined colorimetrically. The increasing rate of PPD oxidation by liver homogenate was higher in tungstate fed animals than in standard diet fed ones. Among the fractionations of liver extract, the mitochondrial and postmitochondrial fractions in the liver extract of tungstate fed animals led to a higher availability of PPD oxidation by PPD treatment compared with standard diet fed ones. In conclusion, these results suggest that an enhanced liver injury in tungstate fed animals treated with PPD may be due to oxygen free radicals produced in other systems except oxygen free radicals generating from cytosolic XO system. Especially, oxidative availability by PPD can be used for oxygen free radical detection in some tissue.

• The Korea Journal of Herbology
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• v.23 no.3
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• pp.53-60
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• 2008

Objectives : Oxidative stress has a critical role in neurodegenerative diseases. In this study, we investigated the antioxidant and neuroprotective effects of the ethanolic extract of Banryong-hwan (BRHE) in SH-SY5Y cells and primary rat mesencephalic dopaminergic neurons. Methods : To assess the antioxidant effects, we carried out 1,1-diphenyl-2-picrylhydrazyl(DPPH) free radical scavenging assay, 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)(ABTS) radical cation decolorization assay, and determination of total polyphenolic content. We evaluated the effect of BRHE treatment on neuroprotection against 6-hydroxydopamine(6-OHDA) toxicity using thiazolyl blue tetrazolium bromide(MTT) assay, nitric oxide(NO) assay, reactive oxygen species(ROS) assay in SH-SY5Y cells and tyrosine hydroxylase(TH) immunocytochemistry in primary rat mesencephalic dopaminergic neurons. Results : BRHE showed IC50 values of 328.10 ${\mu}g/mL$ and 43.12 ${\mu}g/mL$ in DPPH assay and in ABTS assay, respectively. Total polyphenolic content was 180.76 ${\mu}g/mL$. In SH-SY5Y cells, BRHE significantly attenuated the toxicity induced by 6-OHDA at the concentrations of 25-100 ${\mu}g/mL$ pre- and post- treatment in MTT assay. While 6-OHDA increased the NO and ROS contents, BRHE decreased them in a dose dependent manner. Moreover, in primary dopaminergic neuron culture, BRHE significantly protect-ed the dopaminergic cell loss against 6-OHDA toxicity up to 136% at the concentration of 75 ${\mu}g/mL$. Conclusions : These results demonstrate that BRHE has neuroprotective effect against 6-OHDA induced neurotoxicity through decreasing NO and ROS generation.