• 대한의생명과학회지
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• 제21권4호
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• pp.233-240
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• 2015

Some virulence proteins of Helicobacter pylori, such as vacuolating cytotoxic protein A (VacA) and cytotoxin-associated gene protein A (CagA) have been reported to be causative agents of various gastric diseases including chronic gastritis, gastric ulcer or gastric adenocarcinoma. The expression level of these virulence proteins can be regulated when H. pylori is exposed to the antibacterial agent, cyanidin 3-O-glucoside (C3G) as previously reported. In this study, we analyzed the quantitative change of various virulence proteins including CagA and VacA by C3G treatment. We used 2-dimensional electrophoresis (2-DE) to analyze the quantitative change of representative ten proteome components of H. pylori 60190 ($VacA^+/CagA^+$; standard strain of Eastern type). After 2-DE analysis, spot intensities were analyzed using ImageMaster$^{TM}$ 2-DE Platinum software then each spot was identified using matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) or peptide sequencing using Finnigan LCQ ion trap mass spectrometer (LC-MS/MS). Next, we selected major virulence proteins of H. pylori among quantitatively meaningful ten spots and confirmed the 2-DE results by Western blot analysis. These results suggest that cyanidin 3-O-glucoside can modulate a variety of H. pylori pathogenic determinants.

• Natural Product Sciences
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• 제21권1호
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• pp.30-33
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• 2015

Anthocyanins are water soluble plant pigments which are responsible for the blue, red, pink, violet colors in several plant organs such as flowers, fruits, leaves and roots. In recent years, anthocyanin-rich foods have been favored as dietary supplements and health care products due to diverse biological activities of anthocyanins including antioxidant, anti-allergic, anti-diabetic, anti-microbial, anti-cancer and preventing cardiovascular disease. High-performance countercurrent chromatography (HPCCC) coupled with reversed-phase medium pressure liquid chromatography (RP MPLC) method was applied for the rapid and efficient isolation of cyanidin 3-glucoside (C3G) and peonidin 3-glucoside (P3G) from black rice (Oryza sativa L., Poaceae). The crude black rice extract (500 mg) was subjected to HPCCC using two-phase solvent system composed of tert-butyl methyl ether/n-butanol/ acetonitrile/0.01% trifluoroacetic acid (TBME/B/A/0.01% TFA, 1 : 3 : 1 : 5, v/v, flow rate - 4.5 mL/min, reversed phase mode) to give enriched anthocyanin extract (37.4 mg), and enriched anthocyanin extract was sequentially chromatographed on RP-MPLC to yield C3G (16.5 mg) and P3G (8.7 mg). The recovery rate and purity of isolated C3G were 76.0% and 98.2%, respectively, and those of P3G were 58.3% and 96.3%, respectively. The present study indicates that HPCCC coupled with RP-MPLC method is more rapid and efficient than multi-step conventional column chromatography for the separation of anthocyanins.

• Preventive Nutrition and Food Science
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• 제12권3호
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• pp.148-153
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• 2007

Obesity-induced inflammation plays a crucial role in obesity-related pathologies such as type II diabetes and atherosclerosis. Adipose tissue macrophages and the cell-derived proinflammatory chemokines are key components in augmenting inflammatory responses in obesity. Anthocyanins such as cyanidin and $cyanidin-3-O-{\beta}-D-glucoside$ (C3G) are known to elicit anti-inflammatory activities by suppressing the production of proinflammatory mediators such as tumor necrosis factor alpha and nitric oxide in LPS-stimulated macrophages. In the present study, we investigated whether cyanidin and C3G have the potential to suppress the inflammatory responses of adipose cells. Cyanidin and C3G not only suppressed the migration of RAW 264.7 macrophages induced by mesenteric adipose tissue-conditioned medium, but also inhibited the activation of the cells to produce inflammatory chemokines such as monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-related protein-2 (MRP-2) in a dose-dependent manner. Cyanidin and C3G also inhibited the release of MCP-1 and MRP-2 from adipocytes and/or macrophages. These findings suggest that cyanidin and C3G may suppress the inflammatory responses of adipose tissue in obesity.

• Natural Product Sciences
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• 제21권1호
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• pp.20-24
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• 2015

Cyanidin-3-glucoside (C3G) and cyanidin-3-rutinoside (C3R) were isolated by high-performance countercurrent chromatography (HPCCC) using a two-phase solvent system composed of tert-butyl methyl ether/n-butanol/acetonitrile/water/trifluoroacetic acid (1 : 3 : 1 : 5 : 0.01, v/v) to give pure C3G (34.1 mg) and C3R (14.3 mg) from 1.5 g crude mulberry fruit extract. Using the pure C3G and C3R, a reliable high-performance liquid chromatography (HPLC) method was developed and validated to determine the C3G and C3R contents in mulberry fruit. C3G and C3R were separated simultaneously using an Eclipse XDB-C18 column ($4.6{\times}250mm$ I.D., $5{\mu}m$) coupled with a photodiode array detector (PDA). The gradient elution of the mobile phase consisting of acetonitrile (0.5% formic acid) and water (0.5% formic acid) was applied (1.0 mL/min), and the detection wavelength was 520 nm. The calibration curves of C3G and C3R showed good linearity (both with $r^2=0.9996$) in the concentration range $15.625-500{\mu}g/mL$, and the relative standard deviations (RSD%) of intra- and inter-day variability were in the ranges 2.1 - 8.2% and 4.1 - 17.1%, respectively. The accuracies were ranged 96.5 - 102.6% for C3G and C3R, respectively. The developed HPLC method was used to determine the contents of C3G and C3R in newly harvested mulberry from eight different provinces of Korea.

• 한국육종학회지
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• 제43권3호
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• pp.196-200
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• 2011

'슈퍼자미'는 한국방송통신대학교 농학과에서 2009년도에 육성한 C3G 고함유 흑자색 품종으로 주요특성과 수량성을 요약하면 다음과 같다. 1. 중부평야지 평균 출수기는 8월 25일로 만생종이며, 간장은 75 cm 정도이고, 종피색이 흑자색인 유색메벼 품종이다. 2. 현미천립중은 26.2 g 정도이고, 장폭비는 1.87로 단원형이며, 종피는 흑자색으로 안토시아닌 주색소인 C3G 함량이 '흑진주벼'의 10배 수준으로 색택이 균일한 품종이다. 3. 중부평야지역 보통기 보비재배 2개소에서 평균 629.7 kg/10a 수준으로 '흑진주벼'의 162% 수준이다.

• 한국작물학회지
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• 제53권spc호
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• pp.58-64
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• 2008

Cyanidin-3-glucoside(C3G) 고함유 흑자색미 계통을 육성하기 위해 흑진주, 수원425호, 상해향혈나, 용금1호, C3GHi 품종 및 계통들 간에 완전이면교배에서 얻은 20개 조합 $F_1$의 현미 천립중과 C3G함량에 대한 diallel analysis 및 조합 능력 분석 결과는 다음과 같다. 1. 교배모본의 현미 천립중은 16.6 g(흑진주) - 30.4 g(상해향혈나)이고, 현미 100 g당 C3G 함량은 C3GHi가 흑진주의 5배 정도 높은 1043.5 mg으로 나타났다. 2. 완전이면교배 결과 현미 천립중과 C3G 함량은 비대립 유전자 효과는 나타나지 않았고, 유전자의 상가적 및 우성적 유전효과만 인정되었다. 3. 현미 천립중과 C3G 함량의 유전 양식은 두 형질 모두 상가적 및 우성효과에 의한 부분우성을 보였으며, 평균 우성정도는 각각 0.80 및 0.69였고, 교배모본에서 정의대립 유전자 비율이 높았으며, 우성친의 빈도가 높고, 후대에서는 감소하는 방향으로 작용하였다. 4. 두형질의 상가적 유전효과에 의한 협의의 유전력은 각 각 0.66, 0.70이고, 광의의 유전력은 각각 0.98, 0.98로 높게 나타났다. 5. 현미 천립중에 대한 일반조합능력 효과는 상해향혈나가 가장 높았고, 흑진주가 가장 낮았으며, 흑진주/수원425 호, 수원425호/C3GHi 조합의 특정조합능력 효과가 가장 높았다. 6. C3G함량에 대한 일반조합능력 효과는 C3GHi가 가장 높았고, 상해향혈나가 가장 낮았으며, 흑진주/C3GHi 조합에서 특정조합능력 효과가 가장 낮게 나타났다.

• 한국작물학회지
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• 제43권3호
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• pp.179-183
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• 1998

The cyanidin 3-glucoside (C3G) extracted from pigmented rice seeds in 0.5% TFA (Trifluoro acetic acid) -95% ethanol was separated by High Performance Liquid Chromatography (HPLC). A HPLC system using a Develosil ODS-5 column and 0.1 % TFA-$H_2O$～0.1 % TFA-$CH_3CN$ gradient elution was selected for separation and quantitative determination of C3G. Regression equation obtained for the standard content of C3G pigment was as Y=21.95293$^*$X-14.726771 (r=0.99$^{**}$). Using this method, 326 domestic and introduced collections were evaluated for the C3G content. The Korean bred cultivar 'Heugjinjubyeo', showed highest C3G content (552 mg/100g seed) among the tested cultivars. Among the pigmented rice cultivars ten cultivars were selected for containing a high content of C3G. The content of C3G per 100g seeds was in high order as follows: Heugjinjubyeo (552mg)>Cheng Chang (321mg)>Kilimgeugmi (240mg)>PI160979-2 (224mg)>Hong Shei Lo (221mg)>Heugnambyeo (191 mg)>Mitak =PIl60979-1 (186mg)>Suwon425 (163mg)>Sanghaehyanghyeolla (108mg). The C3G pigment was not detected in the common white rice cultivars.

• The Korean Journal of Physiology and Pharmacology
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• 제15권6호
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• pp.353-361
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• 2011

In this study, cyanidin-3-glucoside (C3G) fraction extracted from the mulberry fruit (Morus alba L.) was investigated for its neuroprotective effects against oxygen-glucose deprivation (OGD) and glutamate-induced cell death in rat primary cortical neurons. Cell membrane damage and mitochondrial function were assessed by LDH release and MTT reduction assays, respectively. A time-course study of OGD-induced cell death of primary cortical neurons at 7 days in vitro (DIV) indicated that neuronal death was OGD duration-dependent. It was also demonstrated that OGD for 3.5 h resulted in approximately 50% cell death, as determined by the LDH release assay. Treatments with mulberry C3G fraction prevented membrane damage and preserved the mitochondrial function of the primary cortical neurons exposed to OGD for 3.5 h in a concentration-dependent manner. Glutamate-induced cell death was more pronounced in DIV-9 and DIV-11 cells than that in DIV-7 neurons, and an application of $50{\mu}M$ glutamate was shown to induce approximately 40% cell death in DIV-9 neurons. Interestingly, treatment with mulberry C3G fraction did not provide a protective effect against glutamate-induced cell death in primary cortical neurons. On the other hand, treatment with mulberry C3G fraction maintained the mitochondrial membrane potential (MMP) in primary cortical neurons exposed to OGD as assessed by the intensity of rhodamine-123 fluorescence. These results therefore suggest that the neuroprotective effects of mulberry C3G fraction are mediated by the maintenance of the MMP and mitochondrial function but not by attenuating glutamate-induced excitotoxicity in rat primary cortical neurons.

• 한국작물학회지
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• 제51권2호
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• pp.173-178
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• 2006

Anthocyanin in blackish purple rice is composed of cyanidin, peonidin, malvidin, pelargonidin and delphinidin flavylium ion and their glucosides, which shows antioxidant activity similar to a-tocoperol. It has been demonstrated that cyanidin-3-glucoside(C3G) of anthocyanin has a high-potency antioxidant activity, and is a prominent in anticancer and antithrombotic activity. In analysis of content of anthocyanin from the genetic source of blackish purple rice, the level of C3G was approximately 80% of total contents of anthocyanins, and showed the highest content in Heugjinjubyeo, of which contains an amount of 500 mg by weight of 100g brown rice in comparison that most of varieties has less than 50mg by weight of 100g brown rice. We found that C3G pigment in blackish purple rice was considerably affected by cultural season, cultivation area, ripening temperature, and weather condition, etc. as well as genetic properties. In cross combination between Heugjinjubyeo and Suwon425, intermediate parent $F_9$ generation which was three times higher or more in C3G content than that of Heugjinjubyeo was bred, of which it contained an amount of 1678 mg by weight of 100g brown rice, and we called it 'C3GHi' Rice. And, we found that the C3GHi rice was more excellent than that of the existing Heugjinjubyeo in anticancer and antithrombotic activity. Methanol extract from Heugjinjubyeo was fractionated by organic solvents in order of n-hexane, $CH_{2}Cl_2$ and n-BuOH, and then oryzafuran, quercetin, vanillic acid and protocatechuic acid and their structures from the n-BuOH fraction were ascertained. Oryzafuran of these compounds was a natural compound found firstly in nature. Quercetin and protocatechuic acid besides new compound oryzafuran showed excellent antioxidant ability to vitamin C. These results suggest that blackish purple rice has very high value as a source of various functional food as well as staple food.

• Food Science and Biotechnology
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• 제15권6호
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• pp.993-996
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• 2006

Cyanidin 3-glucoside (C3G) isolated from black rice was investigated for hepatoprotective effects in HepG2 cells under oxidative stress. When an increase in the production of reactive oxygen species (ROS) was induced by gramoxone, cell viability was drastically decreased by 42%. However, in the presence of C3G, no hepatocytic damage was observed in HepG2 cells treated with gramoxone. C3G was found to manifest a stronger scavenging effect (91%) on superoxide anion radical ($O_2\;^{.-}$) than any of the other natural and synthetic antioxidants. Results suggest that C3G from black rice possesses hepatoprotective effects in vitro, which may be, at least in part, due to $O_2\;^{.-}$ scavenging.