• Fisheries and Aquatic Sciences
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• 제21권6호
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• pp.16.1-16.7
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• 2018

Background: Matrix metalloproteinases (MMPs) are linked with several complications such as metastasis of cancer progression, oxidative stress, and hepatic fibrosis. Brown seaweeds are being extensively studied for their bioactive molecule content against cancer progression. In this context, Sargassum horneri was reported to possess various bioactivities including antiviral, antimicrobial, and anti-inflammatory partly due to its phenolic compound content. Methods: In this study, potential of S. horneri was evaluated through anti-MMP effect in HT1080 fibrosarcoma cells. S. horneri crude extract was fractionated with organic solvents, namely, water ($H_2O$), n-buthanol (n-BuOH), 85% aqueous methanol (85% aq. MeOH), and n-hexane. The non-toxicity of fraction samples (Sargassum horneri solvent-partitioned extracts (SHEs)) was confirmed by cell-viability assay. SHEs were tested for their ability to inhibit MMP enzymatic activity through gelatin digestion evaluation and cell migration assay. Expressions of MMP-2 and MMP-9 and tissue inhibitors of MMP (TIMPs) were evaluated by reverse transcription and Western blotting. Results: All fractions inhibited the enzymatic activities of MMP-2 and MMP-9 according to gelatin zymography. Except $H_2O$ fraction, fractions hindered the cell migration significantly. All tested fractions suppressed both mRNA and protein levels of MMP-2, MMP-9, TIMP-1, and TIMP-2. Conclusion: Overall, current results suggested that S. horneri has potential to be a good source for anti-MMP agents, and further investigations are underway for better understanding of the action mechanism and isolation and elucidation of the bioactive molecules.

• Fisheries and Aquatic Sciences
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• 제21권9호
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• pp.21.1-21.8
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• 2018

In the present study, we first evaluated the melanin inhibitory effect of four crude 70% ethanol extracts separated from soft corals abundantly growing along the seawaters of Jeju Island, South Korea, including Dendronephthya castanea (DC), Dendronephthya gigantea (DG), Dendronephthya puetteri (DP), and Dendronephthya spinulosa (DS). Among the four ethanol extracts, the ethanol extract of DP (DPE) did not possess any cytotoxic effect on B16F10 cells. However, all other three extracts showed a cytotoxic effect. Also, DPE reduced the melanin content and the cellular tyrosinase activity without cytotoxicity, compared to the ${\alpha}-MSH$-stimulated B16F10 cells. Specifically, DPE downregulated the expression levels of tyrosinase and microphthalmia-associated transcription factor by activating the ERK signaling cascade in ${\alpha}-MSH$-stimulated B16F10 cells. Interestingly, the melanin inhibitory effect of DPE was abolished by the co-treatment of PD98059, an ERK inhibitor. According to these results, we suggest that DPE has whitening capacity with the melanin inhibitory effects by activating ERK signaling and could be used as a potential natural melanin inhibitor for cosmeceutical products.

• Parasites, Hosts and Diseases
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• 제36권2호
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• pp.133-142
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• 1998

참굴큰입흡충 (Gymnophalloides seoi)의 병원성을 규명하기 위한 연구의 일환으로 성충의 조효 소에서 단백분해효소를 분리 정제한 후 생화학적 특성을 관찰하였다. 조효소를 0.1 M sodium acetate (pH 4.5)로 투석한 후 원심분리하여 얻은 상층액을 Sephacris S-200 HR column chromatography로 부분 정제한 후 DEAE-Sephacelcolumnchromatography를 실시하여 순수 정 제하였다. SDS-PAGE를 실시하여 각 정제 단계별 시료의 정제도를 확인한 결과 분자량이 40 kDa 인 단일 분획이 관찰되었다. 정제된 효소는 시스테인 단백분해효소의 특이억제제인 L-lorans- epoxysuccinylleucylamido (4-guanidino) butane (E-64)와 iodoacetic acid, 세린 및 시스테인 단백분해효소의 일반 억제제인 leupeptin에 의해 활성이 억제되어 시스테인 단백분해효소임을 확 인하였다. 정제된 효소는 콜라겐, 파이브로넥틴과 같은 세포외 기질을 분해하였으나 헤모글로빈은 분해정도가 낮아 반응 12시간 후에도 단량체 (monomer)와 이합체 (dimer)의 양이 대조군과 큰 차이가 없었으며, IgGEa와 slgAE 거의 분해하지 믓하였다. 따라서, 참굴큰입흡충 성충의 40 kDa 시스테인 단백분해효소는 충체가 숙주 체내에서 기생생환을 하는데 필요한 영양분 섭취에 주로 관여할 것으로 생각되었다.

• Parasites, Hosts and Diseases
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• 제50권3호
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• pp.225-227
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• 2012

The seroepidemiological status of toxoplasmosis was surveyed among the residents of Cheorwon-gun, Gangwon-do by means of ELISA using a crude extract antigen of Toxoplasma gondii. The sera of 1,661 adult residents (866 males and 795 females) were collected and checked for IgG antibody titers, which showed 17.0% positive rate (282 sera). The positive rate was significantly different between the sex; 20.6% for males and 13.1% for females (P<0.05). The positive rates were higher in fifties of males (28.7%) and forties of females (20.0%). This positive rate of toxoplasmosis in Cheorwon-gun residents is regarded as the highest among the surveys of different geographical regions of Korea. This high positive rate may due in part to peculiar geographical locality of the surveyed area near the naturally well preserved demilitarized zone (DMZ) or presumably consumption of the pork imported from high endemic nations. Therefore, it is necessary to study further the epidemiology of toxoplasmosis in Cheorwon-gun.

• 생약학회지
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• 제30권3호
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• pp.255-260
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• 1999

In the previous report, E-kong-san, which is usually used for recovering health in traditional medicine, has been shown to decrease cisplatin induced nephrotoxicity in vivo and in vitro. The significant reduction of E-kong-san on the cisplatin induced nephrotoxicity led us to investigate whether the effect of this water extract was a result of triggering antioxidation. In monkey kidney Vero cells, E-kong-san at $5{\sim}10\;mg/ml$ was able to attenuate 2mM cisplatin-stimulated cell death by 46.8% and 31.8%, respectively. E-kong-san showed strong free radical scavengering activities on 1,1-diphenyl-2-picrylhydrazil (DPPH) radical and xanthine/xanthine oxidase (XOD) generated superoxide anion radical $(O_2^{-.})$. We further studied the effects of E-kong-san on lipid peroxidation in rat liver microsomes induced by enzymatic and nonenzymatic methods. Moreover, E-kong-san exhibited significant inhibition on both ascorbic $acid/Fe^{2+}$ and $ADP/NADPH/Fe^{3+}$ induced lipid peroxidation in rat liver microsomes. Based on these results, we suggest that E-kong-san protects the cisplatin induced cytotoxicity by its antioxidative mechanism.

• 생약학회지
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• 제36권4호통권143호
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• pp.324-331
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• 2005

The roots of Rosa rugosa have been used to treat diabetes mellitus in the folkloric society of Korea. To demonstrate the active component for the rat obesity induced by high fat diet for 6 weeks, the phytochemical fractionation and the pharmacological activity test were performed on this crude drug. It was shown that the methanolic extract and its EtOAc fraction inhibited the weight increase of the rat body, abdominal fat pad and hyperlipidemia at 200 mg/kg dose. Further, the triterpenoids, euscaphic acid and tormentic acid, isolated from R. rugosa roots were active at 30 mg/kg in the same assay. The two components shifted serum total-, HDL, and LDL-cholesterol levels toward the values of the unteated group, suggesting that the active compounds has hypolipidemic effects. The rats fad euscaphic acid and tormentic acid also reduced thiobarbituric acid-reactive substance (TBARS) and hydroxyl radical in the rat blood and increased superoxide dismutase activity compared to the control. TBARS values and carbonyl contest of the hepatic protein were reduced by treatment with the two triterpenoids. Antioxidative enzyme (SOD, glutathione peroxidase, and catalase) activities in hepatic were increased by treatment of rats with the triterpenoids, which suggests that triterpenoids inhibited the reduction of hepatic antioxidative activity caused by high fat diet. Taken together, these results support that euscaphic acid and tormentic acid improve a high fat diet-induced hyperlipidemia via the activation of antioxidative mechanism.

• 한국식품과학회지
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• 제35권6호
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• pp.1216-1220
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• 2003

밤꽃의 항산화효과를 측정하기 위해 밤꽃의 수꽃을 수집하여 메탄올로 추출한 후 추출물의 항산화 효과를 측정하였다. 밤꽃 조추출물의 추출수율은 6.26% 였으며, 총 페놀함량은 0.199 mg/mL 인 것으로 나타났다. DPPH법으로 free radical 소거능을 측정한 결과 밤꽃 추출물이 17.22%로 기존의 항산화제에 비해서는 소거능이 다소 낮은 것으로 측정되었다. SOD 유사활성 측정은 밤꽃 추출물이 65.10%로 BHT(95.70%)와 quercetin(93.29%)에 비해 낮았지만, ascorbic acid(30.30%) 보다는 높은 활성을 나타내었다. Lecithin 산화저해율은 밤꽃 추출물이 51.45%, BHT(67.08%), quercetin(63.46%), ascorbic acid(23.60%), chlorogenic acid(48.76%), tannic acid(50.10%)로 나타났다. 이상의 연구결과 밤꽃 추출물이 DPPH free radical 소거능에서 기존 항산화제에 비해 낮은 활성을 나타냈지만, 시간경과에 따른 활성의 지속성과 50% 이상의 지질과산화 억제효과가 있어 천연항산화제로서의 가능성이 충분히 있는 것으로 추정된다.

• 한국식품과학회지
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• 제18권4호
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• pp.270-277
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• 1986

Ammonium sulfate 염분획과 CM-cellulose chromatography로 Fig latex에서 ficin을 분리하여 다음과 같은 결과를 얻었다. 분리한 crude ficin을 전기영동한 결과 main band 와 3개의 minor band로 나타났으며 main peak의 정제에서 얻은 ficin은 단일 band로 나타났다. 분리한 ficin의 pH 및 열에 대한 영향을 PH7.0, $60^{\circ}C$에서 최적 작용을 나타내었다. 분리한 ficin의 pH에 대한 안정성은 pH $2{\sim}8$로 비교적 넓은 범위로 나타났고 열에 대한 안정성은 $0{\sim}55^{\circ}C$의 범위로 안정성이 나타났다. 분리한 ficin의 분자량은 19500으로 나타났다. 산성 아미노산 21.8%. 염기성 아미노산 3.5%, 중성아미노산74.7%로 나타났으며 cystine과 tryptophan을 제외한 아미노산 잔기수는 174로 나타났다.

• 한국축산식품학회지
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• 제29권2호
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• pp.157-163
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• 2009

This study was conducted for the isolation, purification, and characterization of a protease from Korean pear, to see its proteolytic activity on chicken actomyosin and to find the optimum pH and temperature of activity on chicken actomyosin. The protease was isolated from crude extract of Korean pear by ammonium sulfate precipitation. Further purification was done by DEAE-Sepharose ion-exchange chromatography, Mono-Q and Mini-Q column chromatography. The purified enzyme gave a single protein band on SDS polyacrylamide gel electrophoresis and the molecular weight was found to be 38 kDa. The specific activity of purified enzyme was 34,907 unit/mg with 25 fold purification and the yield was 2%. The purified enzyme incubated with chicken actomyosin showed high activity. The optimum pH and temperature for enzyme activity on chicken actomyosin were 6.5 and $70^{\circ}C$, respectively. A protease was purified from Korean pear for the first time and characterized. It was found to be promising for meat tenderization.

• 동의생리병리학회지
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• 제22권2호
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• pp.350-356
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• 2008

Etiological studies of diabetes and its complications showed that oxidative stress might play a major role. Therefore, many efforts have been tried to regulate free oxygen radicals for treating diabetes and its complications. Cheonghyul-san has been known to be effective for the antidiabetic, antihyperlipidemic and antiobesitic prescription, and composed of four crude herbs. In male ob/ob mouse with severe obesity, hyperinsulinemia, hypergiycemia, hyperlipidemia, the acting mechanisms of Cheonghyul-san were examined. Mice were grouped and treated for 5 weeks as follows. Both the lean (C57/BL6J black mice) and diabetic (ob/ob mice) control groups received standard chow. The experimental groups were fed with a diet of chow supplemented with 7.5, 15 and 30 mg Cheonghyul-san per 1 kg of body weight for 14 days. The effects of Cheonghyul-san extract on the ob/ob mice were observed by measuring the serum levels of glucose, insulin, lipid components, and the kidney levels of reactive oxygen species (ROS), MDA+HAE, GSH and also the enzyme activities involved in polyol pathway. Cheonghyul-san lowered the levels of serum glucose and insulin in a dose dependent manner. Total cholesterol, triglyceride and free fatty acid levels were decreased, while the HDL-cholesterol level was increased, in Cheonghyul-san treated groups. Renal aldose reductase and sorbitol dehydrogenase activities were increased in the ob/ob mice, whereas those were inhibited in the Cheonghyul-san-administered groups. Cheonghyul-san inhibited the generation of ROS in the kidney. Finally, MDA+HAE level was increased and the GSH level was decreased in the ob/ob mice, whereas those were improved in the Cheonghyul-san-administered groups. The results suggested that Cheonghyul-san exerted the antidiabetic and antihyperlipidemic activities by regulating the activities of polyol pathway enzymes, scavenging ROS, regulating the MDA+HAE and GSH levels in the ob/ob mice.