• Title/Summary/Keyword: pH dependent

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In vivo protein expression changes in mouse livers treated with dialyzed coffee extract as determined by IP-HPLC

  • Yoon, Cheol Soo;Kim, Min Keun;Kim, Yeon Sook;Lee, Suk Keun
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.40
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    • pp.44.1-44.17
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    • 2018
  • Background: Coffee extract has been investigated by many authors, and many minor components of coffee are known, such as polyphenols, diterpenes (kahweol and cafestol), melanoidins, and trigonelline, to have anti-inflammatory, anti-oxidant, anti-angiogenic, anticancer, chemoprotective, and hepatoprotective effects. Therefore, it is necessary to know its pharmacological effect on hepatocytes which show the most active cellular regeneration in body. Methods: In order to determine whether coffee extract has a beneficial effect on the liver, 20 C57BL/6J mice were intraperitoneally injected once with dialyzed coffee extract (DCE)-2.5 (equivalent to 2.5 cups of coffee a day in man), DCE-5, or DCE-10, or normal saline (control), and then followed by histological observation and IP-HPLC (immunoprecipitation high performance liquid chromatography) over 24 h. Results: Mice treated with DCE-2.5 or DCE-5 showed markedly hypertrophic hepatocytes with eosinophilic cytoplasms, while those treated with DCE-10 showed slightly hypertrophic hepatocytes, which were well aligned in hepatic cords with increased sinusoidal spaces. DCE induced the upregulations of cellular proliferation, growth factor/RAS signaling, cellular protection, p53-mediated apoptosis, angiogenesis, and antioxidant and protection-related proteins, and the downregulations of NFkB signaling proteins, inflammatory proteins, and oncogenic proteins in mouse livers. These protein expression changes induced by DCE were usually limited to the range ± 10%, suggesting murine hepatocytes were safely reactive to DCE within the threshold of physiological homeostasis. DCE-2.5 and DCE-5 induced relatively mild dose-dependent changes in protein expressions for cellular regeneration and de novo angiogenesis as compared with non-treated controls, whereas DCE-10 induced fluctuations in protein expressions. Conclusion: These observations suggested that DCE-2.5 and DCE-5 were safer and more beneficial to murine hepatocytes than DCE-10. It was also found that murine hepatocytes treated with DCE showed mild p53-mediated apoptosis, followed by cellular proliferation and growth devoid of fibrosis signaling (as determined by IP-HPLC), and subsequently progressed to rapid cellular regeneration and wound healing in the absence of any inflammatory reaction based on histologic observations.

Simple and Rapid Evaluation System for Endosulfan Toxicity and Selection of Endosulfan Detoxifying Microorganism Based on Lumbricus rubellus (Lumbricus rubellus를 이용한 endosulfan의 간편, 신속 독성 평가 및 endosulfan 분해 미생물의 선별)

  • Sohn Ho-Yong;Kim Hong-Ju;Kum Eun-Joo;Lee Jung-Bok;Kwon Gi-Seok
    • Journal of Life Science
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    • v.16 no.1
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    • pp.108-113
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    • 2006
  • To compensate the problems of chemical assay in detoxification of recalcitrant and a practical approach in selection of bioremediation bacteria, a simple and rapid toxicity evaluation system was constructed based on Lumbricus rubellus. Long term-culture and specific equipment are not necessary, and semi-quantitative analysis of toxicity at sub-lethal concentration is possible by measuring of dose-dependent increased yellowish secreted compounds. When the toxicity of endosulfan, its metabolites and structurally related chemicals were measured for 24 h, the results were coincided with previous reports. Toxicity was found in endosulfan, endosulfan sulfate, aldrin, and dieldrin, respectively. Rapid and economic selection of endosulfan-detoxifying bacteria was possible using our system. Klebsiella pneumoniae KE-1, K. oxytoca KE-8 and Pseudomonas sp. KS-2P, reported endosulfan degrading bacteria, ameliorated the endosulfan toxicity, whereas E. coli, B. subtilis and other bacteria failed to protect the toxicity of endosulfan in L. rubellus. Our results suggest that the constructed system is useful to selection of microorganism as well as toxicity evaluation against toxic recalcitrants.

Growth of Landscape Tree Species at Two Planting Densities in a Planting Pilot System for Reclaimed Dredging Areas (임해준설매립지 식물재배공정에서 밀도에 따른 조경수목의 생장)

  • Lee, Deok-Beom;Nam, Woong;Kwak, Young-Se;Jeong, In-Ho;Lee, Sang-Suk
    • Journal of the Korean Institute of Landscape Architecture
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    • v.37 no.2
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    • pp.114-123
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    • 2009
  • To investigate the possible use of plants for landscaping in reclaimed soil, a planting pilot system experiment was performed over the course of four years in reclaimed dredging area with four species: Alnus firma, Alnus hirsuta, Pinus thunbergii, and Pyrachantha angustifolia for 4 years. The physicochemical characteristics of the tested soil showed that it was sandy through coming from a reclaimed dredging area. The average pH of the tested soil was 7.16(slight alkali), and electric conductivity(EC) was relatively low, $294{\mu}S/cm$, even though it came from a saltwater area. To test the effect of planting density vs. phytomass by plant specie from a planting basin, the experiment was designed using four plant species with high and low planting densities over 4 years. The planting conditions of the growth of landscape tree species exhibited growth height as follows: A. hirsuta, A. firma, P. thunbergii, and P. angustifolia, whill the DBH followed the order of A. hirsuta, A. firma, and P. thunbergii. The total phytomass of each plant was higher at low density planting areas than high density planting area in terms of total phytomass production and growth distribution in the reclaimed dredging area. Total phytomass per unit area increased as follows: A. hirsuta, A. firma, P. thunbergii, and P. angustifolia. The total phytomass per each tested plant was 2 times higher in low density planting areas than high density planting areas. Total phytomass per unit area, however, was similar or slighty higher in high density planting areas compared to low density areas. Among the tested plants, A. hirsuta showed the highest phytomass, implying that A. hirsuta adapted very well to the reclaimed area and has the capability of a fast growth, nitrogen fixation tree, and utilizing insoluble nutrients through inoculated root nodule bacteria. The yield of phytomass per individual in low density Alnus species was greater than that of the high density. However, those per unit areas had no difference in the density-dependent planting. The ratio of belowground to aboveground was $0.21{\sim}0.26$. Thus, it could be concluded that the Alnus species are potential candidates for ornamental tree species in reclaimed dredging areas. This study offers baseline data for the use of ornamental tree species in reclaimed dredging areas. Additional research is required for different ornamental species in order to increase phytomass of a planting conditions based on reclaimed dredging areas.

Effect of Asarum sieboldii Extracts on the Growth, Acid Production, Adhesion, and Insoluble Glucan Synthesis of Streptococcus mutans (세신 추출물의 Streptococcus mutans에 대한 성장, 산생성, 부착 및 비수용성 글루칸 합성억제에 미치는 영향)

  • Yu Hyeon Hee;Seo Se Jeong;Kim Yeon Hwa;Lee Heung Soo;Kim Kang Ju;Jeon Byung Hun;You Yong Ouk
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.3
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    • pp.666-671
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    • 2003
  • Dental caries are the most commonly occurring chronic diseases in the dental field. Because of increasing sugar consumption and extension of average human life, these diseases are widely found all over the world as the most typical cause for a person to lose a tooth. Therefore, the development of more effective, substantial and safe preventive agents against dental caries is strongly required. Streptococcus mutans is known as the causative bacterial playing the most important role informing plaque and it is being noticed as major causative bacteria of dental caries. The present study was designed to investigate the effect of Asarum sieboldii Miquel(Aristolochiaceae) extracts on the growth, acid production, adhesion, and insoluble glucan synthesis of Streptococcus mutans(S. mutans). Both methanol and aqueous extracts showed concentration dependent inhibitory activity against the growth and acid production of S. mutans, and produced significant inhibition at the concentration of 100, 1,000 and 2,000 μg/ml compared to the control group(p<0.05 - p<0.01). The extracts markedly inhibited S. mutans adherence to HA treated with saliva, and cell adherence was repressed by more than 50% at the concentration of 10 μg/ml and complete inhibition was observed at the concentration of 2,000 μg/ml. On the activity of glucosyltransferase which synthesizes water insoluble glucan from sucrose, methanol and aqueous extracts showed more than 70% inhibition over the concentration of 1,000 μg/ml. Hence, we conclude that Asarum sieboldii might be a candidate of anticaries agent.

The Biochemical Studies on Stored Soy-sauce (저장(貯藏)간장의 생화학적(生化學的) 연구(硏究))

  • Chang, Chi-Hyun
    • Applied Biological Chemistry
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    • v.9
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    • pp.9-27
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    • 1968
  • Studies were carried out in order to elucidate chemical components and microflora in three types of soy-sauce, 12-year aged soy-sauce prepared by improved method. 7-year aged and 20-year aged soy-sauce prepared by ordinary method. They results are summarized as follows: 1. The followings are found to be the important factors affecting the quality of soy sauce. a. Organic acids, reducing sugars and free amino acids were increased in the course of storage. b. In the aged soy-sauces under study non-volatile organic acid increased while volatile organic acid decreased and the total acidity was dependent only upon tie latter. c. It was found that suit concentration decreased during the storage. 2. The results of investigation of microflora in the stored soy-sauce are shown as follows. Soy-sauce Improved Ordinary Microbe 12-Y. 20-Y. 7-Y. Aerobic bacteria colony/1ml. 6 123 2 Halophilic lactic acid bacteria colony/1ml. 4 6 10 Osmophilic yeast colony/1ml. $828{\times}10^4$ 248 - b. In the stored soy-sauces, aerobic bacteria are incapable of growing due to drop in pH value and the influence of salt concentration. c. Halophilic lactic acid bacteria are incapable of growing due to drop in pH value, even the salt concentrations decreased during the storage. d. Osmophilic yeast are still growing in low pH value and in the decreasing salt concentration during the strage. 3. The results of amino acid analysis by paper partition chromatographic and calorimetric methods are shown as follows. a. Fourteen kinds of amino acid and thirteen amino acids were detected in the soy-sauce of 12-year aged improved soy-sauce and 7-year aged and 20-year aged ordinary one, respectively. b. The contents of aspartic acid, glutamic acid, serine, valine, leucine, lysine, histidine and methionine increased in the 20-year aged ordinary soy-sauce compared to the 7-year aged one. On the other hand those of alanine, tyrosine, phenylalanine and cystine decreased. 4. The results of sugar analysis by paper chromatography are as follows. a. In the 12-year aged improved soy-sauce, galactose, glucose, arabinose, xylose, rhamnose, maltose and an unknown were detected, and their amounts were in the above order except maltose and an unknown. b. Both in the 7 and 12-year aged ordinary soy-sauces, galactose, arabinose, xylose, glucose and rhamnose were detected and the amounts of the sugars were in the above order. c. In the non-aged ordinary soy-sauce, glucose was not detected but detected from 7-year and 20-year aged ordinary soy-sauce. 5. The results of organic acid analysis by paper chromatography were as follows. a. As volatile acids, acetic, propionic and butyric acids were detected in the 7-year aged ordinary soy-sauce. On the other hand in both the 20-year aged ordinary soy-sauce and the 12-year aged improved ones, only acetic acid was abundant while propionic and butyric acids were round in trace. It was found that propionic and butyric acids, as the unpleasant flavor components, decreased during the storage. b. In the ordinary soy-sauce, citric acid were produced during the storage and lack, malic and tartaric acids increased in the course of aging while succinic, glycolic, fumaric and malonic acids were shown to decrease. Glutaric and oxalic acids disappeared. Citric acid was produced also in the improved soy-sauce, but lactic, tartaric, succinic, malic, and glycolic acids decreased, while both malonic and glutaric acids disappeared. From the above results the citric acid production was considered to be a favorable factor for the taste. c. In the aged soy-sauces, pyruvic, α-ketoglutaric and probably acetoacetic and oxaloacetic acids (both in trace) were present and their amounts were in the above order. All of the α-keto acid abruptly decreased during the storage.

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Melanogenesis Inhibitory and Antioxidant Activities of Phellinus baumii Methanol Extract (장수진흙버섯 메탄올 추출물의 멜라닌 생성 저해작용)

  • Lee, J.S.;Shin, D.B.;Lee, S.M.;Kim, S.H.;Lee, T.S.;Jung, D.C.
    • The Korean Journal of Mycology
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    • v.41 no.2
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    • pp.104-111
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    • 2013
  • Phellinus baumii is a medicinal mushroom used in Asian countries for a long period of time. The purpose of this study was to investigate the skin whitening activities of methanol extracts from fruiting bodies of P. baumii. To evaluate the antioxidant activities of the extract, polyphenol and flavonoid contents, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity and chelating activity on ferrous ions were studied. For assay of skin whitening activities, tyrosinase and DOPA inhibitory activities, and tyrosinase and melanin synthesis inhibitory activities of B16/F10 melanoma cells treated with the methanol extract were investigated. The total polyphenol content of P. baumii methanol extract was 4.19. DPPH scavenging ability of P. baumii methanol extract was 88.26% in $25{\mu}g/mL$ concentration. We tested tyrosinase inhibitory activity and melanin contents in melanoma cells. The tyrosinase activity was inhibited to 65.17% at the concentration of $125{\mu}g/mL$ and melanin synthesis was inhibited to 57.61% at the concentration of $25{\mu}g/mL$. Overall, the experimental results showed that P. baumii methanol extract had inhibitory activities of tyrosinase and melanin synthesis by dose dependent manner in B16/F10 melanoma cells. Strong ultra-violet absorption spectra in the range of 270~370 nm indicated that ethanol extract of P. baumii could protect the skin from UV. Therefore, P. baumii methanol extract might be used for development of skin whitening, anti-UV and skin care agents.

Effects of Adenosine on the Action Potentials of Rabbit SA Nodal Cells (동방결절 활동전압에 대한 아데노신 효과)

  • Kim, Ki-Whan;Ho, Won-Kyung
    • The Korean Journal of Physiology
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    • v.18 no.1
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    • pp.19-35
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    • 1984
  • Since the first report of Drury and $Szent-Gy{\ddot{o}}rgyi$ in 1929, the inhibitory influences of adenosine on the heart have repeatedly been described by many investigators. These studies have shown that adenosine and adenine nucleotides have overall depressant effects, similar to those of acetylcholine. Heart beats become slow and weak. It is also well known that adenosine is a potent endogenous coronary vasodilator. Many investigations on the working mechanisms of adenosine have been focused mainly on the effects of the coronary blood flow. However, the cellular mechanisms underlying the inhibitory action of adenosine on sinus node are not well understood yet. Thus, this study was undertaken to examine the behavior of rabbit SA node under influence of adenosine. In these series of experiments three kinds of preparations were used: whole atrial pair, left atrial strip, and isolated SA node preparations. The electrical activity of SA node was recorded with conventional glass microelectrodes 30 to 50 $M{\Omega}$. The preparations were superfused with bicarbonate-buffered Tyrode solution of pH 7.35 and aerated with a gas mixture of $3%\;CO_2-97%\;O_2$ at $35^{\circ}C$. In whole atrial pair, adenosine suppressed sinoatrial rhythm in a dose-dependent manner. Effect of adenosine on atrial rate appeared at the concentration of $10^{-5}M$ and was enhanced in parallel with the increase in adenosine concentration. Inhibitory action of adenosine on pacemaker activity was more prominent in the preparation pretreated with norepinephrine, which can steepen the slope of pacemaker potential by increasing permeability of $Ca^{+2}$. Calcium ions in perfusate slowly produced a marked change in sinoatrial rhythm. Elevation of the calcium concentration from 0.3 to 8 mM increased the atrial rate from 132 to 174 beats/min, but over 10 mM $Ca^{+2}$ decreased. The inhibitory effect of adenosine on sinoatrial rhythm developed very rapidly. Atrial rate was recovered promptly from the adenosine-induced suppression by the addition of norepinephrine, but extra $Ca^{+2}$ was less suitable to restore the suppression of atrial rate. Adenosine suppressed also atrial contractility in the same dosage range that restricted pacemaker activity, even in the reserpinized preparation. In isolated SA node preparation, spontaneous firing rate of SA node at $35^{\circ}C$(mean{\pm}SEM, n=16) was $154{\pm}3.3\;beats/min. The parameters of action potentials were: maximum diastolic potential(MDP), $-73{\pm}1.7\;mV: overshoot(OS), $9{\pm}1.4\;mV: slope of pacemaker potential(SPP), $94{\pm}3.0\;mV/sec. Adenosine suppressed the firing rate of SA node in a dose-dependent manner. This inhibitory effect appeared at the concentration of $10^{-6}M$ and was in parallel with the increase in adenosine concentration. Changes in action potential by adenosine were dose-dependent increase of MDP and decrease of SPP until $10^{-4}M$. Above this concentration, however, the amplitude of action potential decreased markedly due to the simultaneous decrease of both MDP and OS. All these effects of adenosine were not affected by pretreatment of atropine and propranolol. Lowering extra $Ca^{2+}$ irom 2 mM to 0.3 mM resulted in a marked decrease of OS and SPP, but almost no change of MDP. However, increase of perfusate $Ca^{2+}$ from 2 mM to 6 or 8 mM produced a prominent decrease of MDP and a slight increase of OS and SPP. Dipyridamole(DPM), which is known to block the adenosine transport across the cell membrane, definately potentiated the action of adenosine. The results of this experiment suggest that adenosine suppressed pacemaker activity and atrial contractility simultaneously and directly, by decreasing $Ca^{2+}-permeability$ of nodal and atrial cell membranes.

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Anti-inflammatory Activity of Crinum asiaticum Linne var. Japonicum Extract and its Application as a Cosmeceutical Ingredient (문주란의 항염효과와 화장료적 특성)

  • Kim, Ki-Ho;Kim, Young-Heui;Kim, Ki-Soo;Park, Sun-Hee;Lee, Soo-Hee;Kim, Young-Jin;Kim, Young-Sil;Kim, Jong-Heon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.32 no.1 s.55
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    • pp.59-64
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    • 2006
  • Crinum asiaticum Linne var. japonicum has long been used as a rheumatic remedy, an anti-pyretic, an anti-ulcer treatment, and for the alleviation of local pain and fever in Korea and Malaysia. In order to investigate the possibility of Crinum asiaticum Linne var. japonicum extract as a cosmetic ingredient, we measured its anti-inflammatory effect by inhibition of iNOS (inducible nitric oxide synthase), and the release of PGE2, IL-6, and IL-8. HPLC experiment after extraction with 95% ethanol at pH 3.5 showed that Crinum asiaticum Linne var. japonicum was mainly composed of lycorine (up to 1%), a well-known immunosuppressant. The content of lycorine varied depending on the type of tissue analyzed and the extraction method. In anti-inflammatory assay for inhibition of nitric oxide formation on lipopolysaccharide (LPS)- activated mouse macrophage RAW 264.7 cells, the ethanolic extract of Crinum asiaticum showed inhibitory activity of NO production in dose-dependent manner ($IC_{50} = 83.5 {\mu}g/mL$). Additional study by RT-PCR demonstrated that the extract of Crinum asiaticum significantly suppressed the expression of the iNOS gene. Moreover, the extract of Crinum asiaticum did not show my cytotoxicity, but did show cell proliferation effect against LPS ($10{\sim}60%$ increase of tell viability). In an assay to determine inhibition of the $H_2O_2$-activated release of PGE2, IL-6, and IL-8 in human normal fibroblast cell lines, the release of PGE2 and IL-6 was almost completely inhibited above concentrations of 0.05% and 1%, respectively. Moreover, the release of IL-8 was completely inhibited over the entire range of concentrations (> 0.0025%). The result showed that the extract of Crinum asiaticum Linne var. japonicum has sufficient anti-inflammatory effect. There-fore, Crinum asiaticum Linne var. japonicum extract may be useful as an ingredient of cosmetic products.

Effects of Fermentation on the Metabolic Activities of Pine Needle Juice (발효과정이 솔잎 착즙액의 항산화, alpha-Glucosidase 및 Angiotensin Converting Enzyme 저해 활성에 미치는 영향)

  • Kim, So-Yun;Lee, Hyun-Jung;Park, Jae-Hee;Kim, Rae-Young;Cheong, Hyeonsook;Park, Eunju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.3
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    • pp.325-334
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    • 2013
  • The objective of this study was to compare the content and metabolic activities between fresh pine needle juice (PNJ) and fermented pine needle juice (FPNJ). A variety of factors were measured, including total phenolic content (TPC), antioxidant activity [DPPH radical scavenging activity (RSA), total radical-trapping antioxidant potential (TRAP), oxygen radical absorbance capacity (ORAC), cellular antioxidant capacity (CAC)], anti-genotoxic activity, ${\alpha}$-glucosidase inhibitory activity, and angiotensin converting enzyme (ACE) inhibitory activity. The TPC was $17.3{\pm}0.2$ and $4.6{\pm}0.0$ mg GAE/g in PNJ and FPNJ, respectively. The DPPH RSA, TRAP, and ORAC values increased in a dose-dependent manner for both PNJ and FPNJ, with significantly higher activities in PNJ than FPNJ. The CAC against AAPH-induced oxidative stress in HepG2 cells was protected by both PNJ and FPNJ. Pretreatment with PNJ and FPNJ in human leukocytes produced significant reductions in $H_2O_2$-induced DNA damage at a concentration of $50{\mu}g/mL$. ${\alpha}$-Glucosidase inhibitory activity was significantly higher in FPNJ than PNJ. The ACE inhibitory activity was about 87.1% and 60.0% in 1:1 diluted PNJ and FPNJ, respectively. This study suggests that the fermentation of PNJ could enhance the regulation of blood glucose metabolism and both PNJ and FPNJ might be a new potential source of natural antioxidant, anti-diabetic, and anti-hypertensive agents applicable to food.

Physiological Activity of Sarcodon aspratus Extracts (능이버섯(Sarcodon aspratus) 추출물의 생리활성)

  • 송재환;이현숙;황진국;한정환;노정근;금동혁;박기문
    • Food Science of Animal Resources
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    • v.23 no.2
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    • pp.172-179
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    • 2003
  • This study was carried out to find the preventive medical and therapeutic effects of Sarcodon asparatus on adult disease by employing several biological and biochemical assays. Nitrate scavenging ability(NSA) of Sarcodan asparatus extracts was displayed up to 99.9% at pH 1.2 in a dose-dependent manner. They also had 90.4% electron donating ability(EDA) at the concentration of 0.1 mg/mL. Extracts of Sarcodon asparatus were also able to function as a powerful antioxidant at all concentrations(0.01∼l.0 mg/mL). Furthermore, we observed that 1 mg/mL concentration of the extracts was more powerful than BHT, With respect to fibrolytic activity, Sarcodon asparatus showed 1,843.8 unit/g, which was higher than streptokinase(1,189 unit/g). The inhibitory effects of the extracts on angiotensin converting enzyme, measured by the normal and pretreatment methods, were 53 and 58%, respectively. We also performed cytotoxicity effect of Sarcodon asparatus extracts on a various cancer cell lines. The growth inhibitory effects of the extracts(5.0 mg/mL) on A549, HeLa, AGS, and SK-Hep-1 cells were 78.9, 55.3, 69.0, and 42.5 %, respectively. Interestingly, Sarcodon asparatusextracts induced mutation on Salmonella typhimurium TA98 and TA100 when Ames test was done.