• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.7
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• pp.1029-1035
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• 2013

The objective of this study was to examine the ability of extracts from Phellinus linteus (PL) and rice with Phellinus linteus mycelium (PLM) to inhibit obesity and diabetes. The efficacy of PL and PLM were evaluated using Oil Red O staining, cholesteryl ester transfer protein (CETP) levels, protein tyrosine phosphate 1B (PTP1B) levels, organ weight, and serum lipid levels. Lipid accumulation significantly decreased by 76% and 59% upon treatment with $300{\mu}g/mL$ of PL and PLM, respectively (P<0.01). The inhibition of CETP activity increased 99% upon treatment with $300{\mu}g/mL$ of PL or PLM. Treatment with 3, 10, 30, 100, and $300{\mu}g/mL$ of PL, changed PTP1B activity by 10, 11, 14, 12, and 18% respectively. Also, treatment with increasing concentrations of PLM led to a significant concentration-dependent inhibition of PTP1B activity (P<0.01). PL and PLM were orally administered for 28 days after a high fat diet (HFD). PL significantly (P<0.05) reduced triglyceride and cholesterol levels. In addition, PLM significantly (P<0.05) reduced triglyceride, cholesterol, and HDL-cholesterol levels. GOT and GPT were not significantly affected. These results indicate that PL and PLM extracts have potent and useful activities for the treatment of obesity and diabetes mellitus.

• Applied Biological Chemistry
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• v.41 no.6
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• pp.414-420
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• 1998

We studied the difference effects of dietary ${\alpha}-linolenic\;acid\;({\alpha}-LA,\;18:3\;n-3)$, eicosapentaenoic acid (EPA, 20:5 n-3) and docosahexaenoic acid (DHA, 22:6 n-3) on the lowering of triacylglycerol in the liver and serum on lipid metabolism in rats. Rats were fed semipurified diets containing 10% fat with constant polyunsaturated/monounsaturated/saturated fatty acids (1:1:1) and n-6/n-3 ratio (1:2). EPA (98%) and DHA (98%) were added in diets as the ethyl esters. The concentration of liver triacylglycerol was significantly lower in rats fed both EPA and DHA than in those fed ${\alpha}-LA$. The concentration of liver phospholipid was significantly higher in rats fed DHA than in those fed ${\alpha}-LA$ and EPA. Both EPA and DHA reduced serum triacylglycerol concentration compared with ${\alpha}-LA$, but this effect was more pronounced in the EPA diet. The activity of phophatidate phosphohydrolase in the liver microsome was significantly lower in rats fed both EPA and DHA than in those fed ${\alpha}-LA$. but, there was no significant difference on the activities of diacylglycerol acyltransferase among the three groups. The concentration of liver triacylglycerol were correlated with changes in the microsomal phosphatidate phosphohydrolase activity (r=0.84). Hepatic NADPH generating enzyme, glucose-6-phosphate dehydrogenase was more effective to reduce the activity in rats fed both EPA and DHA than in those fed ${\alpha}-LA$. In conclusion, EPA or DHA reduced the hepatic triacylglycerol concentration by inhibiting microsomal phosphatidate phosphohydrolase, thereby inhibiting synthesis of triacylglycerol in the liver.

• The Korean Journal of Physiology and Pharmacology
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• v.14 no.3
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• pp.177-183
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• 2010

Tetrahydrobiopterin (BH4), an essential cofactor for nitric oxide synthase (NOS) activity, is known to play important roles in modulating both NO and superoxide production during vascular diseases such as atherosclerosis. However, the role of BH4 in functions of vascular smooth muscle cells is not fully known. In this study, we tested the effects of BH4 and dihydrobiopterin (BH2), a BH4 precursor, on migration and proliferation in response to platelet-derived growth factor-BB (PDGF-BB) in rat aortic smooth muscle cells (RASMCs). Cell migration and proliferation were measured using a Boyden chamber and a 5-bromo-2'-deoxyuridine incorporation assay, respectively, and these results were confirmed with an ex vivo aortic sprout assay. Cell viability was examined by 2,3-bis [2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide assays. BH4 and BH2 decreased PDGF-BBinduced cell migration and proliferation in a dose-dependent manner. The inhibition of cell migration and proliferation by BH4 and BH2 was not affected by pretreatment with $N^G$-nitro-L-arginine methyl ester, a NOS inhibitor. Moreover, the sprout outgrowth formation of aortic rings induced by PDGF-BB was inhibited by BH4 and BH2. Cell viability was not inhibited by BH4 and BH2 treatment. The present results suggest that BH4 and BH2 may inhibit PDGF-stimulated RASMC migration and proliferation via the NOS-independent pathway. Therefore, BH4 and its derivative could be useful for the development of a candidate molecule with an NO-independent anti-atherosclerotic function.

• Natural Product Sciences
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• v.13 no.1
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• pp.33-39
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• 2007

Eleven polyhydroxyursane triterpenoids (PHUTs) were tested to determine their cytoprotective, immunosuppressive and anti-inflammatory effects. To compare the bioactivities of $19{\alpha}$-hydroxyursane-type triterpenoids {23-hydroxytormentic acid (6), its methyl ester (7), tormentic acid (8), niga-ichigoside $F_1$ (9),euscaphic acid (10) and kaji-ichigoside $F_1$ (11)} of the Rosaceae crude drugs (Rubi Fructus and Rosa rugosae Radix) with PHUTs possessing no $19{\alpha}-hydroxyl$ of Centella asiatica (Umbelliferae), the four PHUTs, asiaticoside (1), madecassoside (2), asiatic acid (3), and madecassic acid (4) were isolated from C. asiatica and 23-hydroxyursolic acid (5) from Cussonia bancoensis. Cytoprotective effects were assessed by measuring cell viabilities against cisplatin-induced cytotoxocity in $LLC-PK_1$, cells (proximal tubule, pig kidney) to determine whether these agents have protective effects against nephrotoxicity caused by cisplatin. The inhibitory effect of 11 PHUTS on nitric oxide (NO) and prostaglandin $E_2\;(PGE_2)$ were evaluated by measuring nitrite accumulation in lipopolysaccharide (LPS)-induced macrophage RAW 264.7 cells, and their anti-inflammatory effects were tested in 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced mouse ear edema model. Six MHUTs (compounds 1, 2, 4, 6, 10, and 11) exhibited higher cell viabilities during cisplatin-induced cytotoxicity testing even at a concentration of $200\;{\mu}g/ml$ than cisplatin only-treated group, suggesting that ese compounds have the potentcytoprotective efffcts. Compounds 1 and 3 of the C. asiatica and niga-ichigoside $F_1$ exhibited no inhibitory effect on NO and/or $PGE_2$ production whereas other PHUTs produced mild to significant NO and/or $PGE_2$ production.The four compounds (2, 5, 9, and 10) potently inhibited mouse ear edema induced by TPA whereas two compounds (1 and 3) had no activity in this test. These results suggest that many PHUTs are potentchemopreventives. Structure-activity relationship (SAR) was also discussed in each assay with regard to the significant role of OHs at the position of 2, 3, 6, 19, and 23 and to the glycoside linkage at the 28-carboxyl.

• Korean Journal of Food Science and Technology
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• v.30 no.3
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• pp.638-643
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• 1998

The volatile flavor components in Takju made from mashed glutinous rice and barley rice were identified by GC and GC-MS using polar column. Twenty eight kinds of flavor compounds including 10 alcohols, 9 esters, 2 acids, 7 aldehydes were identified. At the beginning of fermentation 14 kinds of volatile components were detected and the kinds of volatile components were increased up to maximum after fermentation for 16 days. The contents of volatile components in the sample made from glutinous rice contained slightly higher than the sample made from barley rice. Ethyl alcohol, n-propyl alcohol, iso-amyl alcohol, n-hexyl alcohol, hexenyl alcohol, 2-phenylethyl alcohol, ethyl acetate, ethyl succinate, 2-phenylethyl acetate, butyic acid, benzaldehyde and 3-methoxybenzaldehyde were detected in the both of the sample throughout the fermentation process. Five kinds of aldehyde including ethyl propionate, iso-amyl acetate, ethyl caprylate and ethylphenyl acetate were detected only in the sample fermented for 16 days. The main components of the both sample were ethyl alcohol, iso-amyl alcohol, 2-phenylethyl alcohol, 2-phenylethyl acetate, hexenyl alcohol and iso-butyl alcohol. Besides ethyl acetate and benzaldehyde from glutinous rice Takju and hexenyl alcohol from barley rice Takju were found as main components.

• Natural Product Sciences
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• v.27 no.2
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• pp.86-98
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• 2021

This study was designed to characterize the effect of ginsenoside-Rg2 (Rg2), one of panaxatriol saponins isolated from Korean ginseng root, on the release of catecholamines (CA) in the perfused model of the rat adrenal medulla, and also to establish its mechanism of action. Rg2 (3~30 µM), administered into an adrenal vein for 90 min, depressed acetylcholine (ACh)-induced CA secretion in a dose- and time-dependent manner. Rg2 also time-dependently inhibited the CA secretion induced by 3-(m-chloro-phenyl-carbamoyl-oxy)-2-butynyltrimethyl ammonium chloride (McN-A-343), 1.1-dimethyl-4-phenyl piperazinium iodide (DMPP), and angiotensin II (Ang II). Also, during perfusion of Rg2, the CA secretion induced by high K⁺, veratridine, cyclopiazonic acid, methyl-1,4-dihydro-2,6-dimethyl-3-nitro-4-(2-trifluoro-methyl-phenyl)-pyridine-5-carboxylate (Bay-K-8644) depressed, respectively. In the simultaneous presence of Rg2 and N^ω-nitro-L-arginine methyl ester hydrochloride ʟ-NAME), the CA secretion induced by ACh, Ang II, Bay-K-8644 and veratridine was restored nearly to the extent of their corresponding control level, respectively, compared to those of inhibitory effects of Rg2-treatment alone. Virtually, NO release in adrenal medulla following perfusion of Rg2 was significantly enhanced in comparison to the corresponding spontaneous release. Also, in the coexistence of Rg2 and fimasartan, ACh-induced CA secretion was markedly diminished compared to the inhibitory effect of fimasartan-treated alone. Collectively, these results demonstrated that Rg2 suppressed the CA secretion induced by activation of cholinergic as well as angiotensinergic receptors from the perfused model of the rat adrenal gland. This Rg2-induced inhibitory effect seems to be exerted by reducing both influx of Na⁺ and Ca²⁺ through their ionic channels into the adrenomedullary cells as well as by suppressing Ca²⁺ release from the cytoplasmic calcium store, at least through the elevated NO release by activation of NO synthase, which is associated to the blockade of neuronal cholinergic and AT₁-receptors. Based on these results, the ingestion of Rg2 may be helpful to alleviate or prevent the cardiovascular diseases, via reduction of CA release in adrenal medulla and consequent decreased CA level in circulation.

• Applied Biological Chemistry
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• v.42 no.3
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• pp.229-234
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• 1999

The effects of linoleic acid(LA, 18 : 2) and/or bovine serum albumin(BSA) on the lipid metabolism in human hepatoma cell line HepG2 cells were evaluated. HepG2 cells were cultured in basal Dulbecco's modified Eagle's(DME) medium(Basal medium), DME medium containing 0.2 mM LA(LA medium), or DME medium containing both 0.2 mM LA and 0.2-1.0% BSA(LA+BSA medium). $[^{14}C]Acetate(0.3\;{\mu}Ci/ml\;medium)$ was added as a radioactive lipid precursor and the cells were incubated for 6 hours. An addition of LA to basal medium resulted in a decrease in the incorporation of $[^{14}C]acetate$ into total cholesterol fraction. In contrast, an addition of BSA to LA-containing medium tended to increase the incorporation of $[^{14}C]acetate$ into total cholesterol. The alteration of cholesterol metabolism in HepG2 cells incubated in LA+BSA medium was attributed by an increase in the incorporation of $[^{14}C]acetate$ into free cholesterol, but not cholesteryl ester fraction. In addition, the secretion of cholesterol was increased by LA+BSA medium, suggesting that BSA stimulates cholesterol secretion. No significant change in the incorporation of $[^{14}C]acetate$ into cellular total lipids was observed among the experimental groups. However, an increased incorporation of $[^{14}C]-labelled$ fatty acid into cellular triacylglycerol and decreased incorporation into phospholipid were observed in cells incubated with LA+BSA medium as compared to those of LA medium. The secretions of $[^{14}C]-labelled$ triacylglycerol, phospholipid, and free fatty acid were also stimulated in HepG2 cells incubated with LA+BSA medium. In conclusion, the present study suggests that in human hepatocytes, LA and BSA influence lipid metabolism, and BSA enhances the secretion of lipids.

• The Korean Journal of Nuclear Medicine
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• v.33 no.6
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• pp.527-536
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• 1999

Purpose: Radiotracers that bind to the central benzodiazepine receptor are useful for the investigation of various neurological and psychiatric diseases. [C-11]Flumazenil, a benzodiazepine antagonist, is the most widely used radioligand for central benzodiazepine receptor imaging by PET. We synthesized 3-(2-[F-18]fluoro)flumazenil, a new fluorine-18 ($t_{1/2}$= 110 min) labeled analogue of benzodiazepine receptor imaging agent, and evaluated in vivo for biodistribution in mice. Materials and Methods: Flumazenil (Ro 15-1788) was synthesized by a modification of the reported method. Precursor of 3-(2-[F-18]fluoro)flumazenil, the tosylated flumazenil derivative was prepared by the tosylation of the ethyl ester by ditosylethane. [F-18] labeling of tosyl substitued flumazenil precursor was performed by adding F-18 ion at $85^{\circ}C$ in the hot ceil for 20 min. The reaction mixture was trapped by C18 cartridge, washed with 10% ethanol, and eluted by 40% ethanol. Bidistribution in mice was determined after intravenous injection. Results: The total chemical yield of tosylated flumazenil derivative was ${\sim}40%$. The efficiency of labeling 3-(2-[F-18]fluoro)flumazenil was 66% with a total synthesis time of 50 min. Brain uptakes of 3-(2-[F-18]fluoro)flumazenil at 10, 30, 60 min after injection, were $2.5{\pm}0.37,\;2.2{\pm}0.26,\;2.1{\pm}0.11$ and blood activities were $3.7{\pm}0.43,\;3.3{\pm}0.07,\;3.3{\pm}0.09%ID/g$, respectively. Conclusion: We synthesized a tosylated flumazenil derivative which was successfully labeled with no-carrier-added F-18 by nucleophilic substitution.

• Natural Product Sciences
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• v.20 no.3
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• pp.160-169
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• 2014

44 compounds and 9 minerals were isolated from and detected in the New Zealand deer velvet antler Cervus elaphus var. scoticus L$\ddot{o}$nnberg. The chemical structures of (1 - 26) were identified on the basis of the spectroscopic methods and comparisons with literature, respectively. The structures were identified as cholesterol (CS, 6), 7-keto-CS (7), $7{\beta}$-hydroxy-CS (8), and $7{\alpha}$-hydroxy-CS (9), and included 12 steroid $3{\beta}$-O-(palmitic/stearic/myristic acid esters; PM/SA/MS) [CS-$3{\beta}$-O-PM (1 - 1), CS-$3{\beta}$-O-SA (1 - 2), CS-$3{\beta}$-O-MR (1 - 3), 7-keto-CS-$3{\beta}$-O-PM (2 - 1), 7-keto-CS-$3{\beta}$-O-SA (2 - 2), 7-keto-CS-$3{\beta}$-O-MR (2 - 3), $7{\beta}$-hydroxy-CS-$3{\beta}$-O-SA (3 -1), $7{\beta}$-hydroxy-CS-$3{\beta}$-O-PM (3 - 2), $7{\beta}$-hydroxy-CS-$3{\beta}$-O-MR (3 - 3), $7{\alpha}$-hydroxy-CS-$3{\beta}$-O-SA (4 - 1), $7{\alpha}$-hydroxy-CS-$3{\beta}$-O-PM (4 - 2), and $7{\alpha}$-hydroxy-CS-$3{\beta}$-O-MR (4 - 3)], dinonyl phthalate (5), 8 nucleic acids analogues [uracil (10), deoxyguanosine (11), deoxyuridine (12), uridine (13), deoxyadenosine (14), adenosine (15), inosine (16), and guanosine (17)], and the 9 free amino acids [L-phenylalanine (18), L-isoleucine (19), L-leucine (20), L-tyrosine (21), L-valine (22), L-proline (23), L-threonine (24), L-alanine (25), and L-hydroxyproline (26)]. Also, there are 8 kinds of amino acids [asparagine, serine, glutamine, glycine, histidine, arginine, methionine, and lysine], 2 sialic acids [N-acetylneuraminic acid (27), ketodeoxynonulosonic acid (28)], and 9 minerals [Na > K > Ca > Mg > Fe > Zn > B > Al > Cu] were detected from the autoaminoacid analyzer and ICP spectrometer, HPAEC-PAD/HPLC-FLD, respectively. 9 kinds of oxycholesterol-$3{\beta}$-O-fatty acid ester (2 - 1, 2 - 2, 2 - 3, 3 - 1, 3 - 2, 3 - 3, 4 - 1, 4 - 2, and 4 - 3) and 3 nucleic acids (12, 14, and 15) were isolated from the velvet antler for the first time. 6 kinds of steroids (7, 8, 9, 2 - 1, 3 - 1, and 4 - 1) were examined for their anti-proliferative effects against L1210, P388D1, K562, MEG-01, KG-1, MOLT-4, A549, HepG2, MCF-7, SK-OV-3, and SW-620 cancer cell lines. They showed anti-proliferative effects with $IC_{50}$ values of 0.06, 2.16, 2.42, > 50.0, 1.66 and $8.31{\mu}M$ against L1210, while the values were 24.05, 9.44, 5.22, 0.25. 9.48 and $49.77{\mu}M$ against P388D1, respectively. The others were inactive.

• Korean Journal of Microbiology
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• v.52 no.2
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• pp.192-201
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• 2016

Recently, Psychrobacter sp. ArcL13 strain showing the extracellular lipase activity was isolated from the Chuckchi Sea of the Arctic Ocean. However, due to the low expression levels of the enzyme in the natural strain, the production of recombinant lipase is crucial for various applications. Identification of the gene for the enzyme is prerequisite for the production of the recombinant protein. Therefore, in the present study, a novel lipase gene (ArcL13-Lip) was isolated from Psychrobacter sp. ArcL13 strain by gene prospecting using PCR, and its complete nucleotide sequence was determined. Sequence analysis showed that ArcL13-Lip has high amino acid sequence similarity to lipases from bacteria of some Psychrobacter genus (84-90%) despite low nucleotide sequence similarity. The lipase gene was cloned into the bacterial expression plasmid and expressed in E. coli. SDS-PAGE analysis of the cells showed that ArcL13-Lip was expressed as inclusion bodies with a molecular mass of about 35 kDa. Refolding was achieved by diluting the unfolded protein into refolding buffers containing various additives, and the highest refolding efficiency was seen in the glucose-containing buffer. Refolded ArcL13-Lip showed high hydrolytic activity toward p-nitrophenyl caprylate and p-nitrophenyl decanoate among different p-nitrophenyl esters. Recombinant ArcL13-Lip displayed maximal activity at $40^{\circ}C$ and pH 8.0 with p-nitrophenyl caprylate as a substrate. Activity assays performed at various temperatures showed that ArcL13-Lip is a cold-active lipase with about 40% and 73% of enzymatic activity at $10^{\circ}C$ and $20^{\circ}C$, respectively, compared to its maximal activity at $40^{\circ}C$.