• Journal of Plant Biotechnology
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• 제40권4호
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• pp.217-223
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• 2013

국내 자생하는 야생칡(Pueraria lobata)은 푸에라린, 다이드진, 다이드제인 등 다양한 이소플라본을 함유하고 있는 주요한 콩과 약용작물이다. 이들 이소플라본은 장내에서 특정 세균과 반응으로 생물전환이 일어나 디하이드로다이드제인, 테트라하이드로다이드제인을 거쳐 약리작용이 타 이소플라본 보다 월등한 이퀄로 물질대사가 이루어지는 것으로 알려져 있다. 본 연구에서는 국내 야생칡으로부터 유도된 캘러스를 이용하여 배양배지, 배양조건 및 배양기간에 따른 캘러스의 증식과 이소플라본의 합성 정도를 구명하고 배양된 캘러스 추출물을 생물전환 능력이 있는 것으로 밝혀진 Pediococcus pentosaceus 균주와의 반응을 통하여 이퀄 전구체인 디하이드로다이드제인의 생합성을 구명하고자 수행되었다. 캘러스의 증식은 광조건에서 1.0 mg/L TDZ과 1.0 mg/L NAA가 첨가된 배지에서 12일 배양했을 때 가장 좋다. 푸에라린 함량은 암조건에서 2.0 mg/L kinetin과 1.0 mg/L NAA가 첨가된 배지에서 16일 배양했을 때 가장 높게 나타났으나, 다이드진과 다이드제인 함량은 처리에 따른 유의한 차이가 나타나지 않았다. 캘러스 추출물을 P. pentosaceus 균주와 반응하여 발효시켰을 때 생물전환에 의해 이퀄의 전구체인 디하이드로다이드제인이 성공적으로 생합성되는 것이 확인되었다. 본 연구의 결과는 국내 야생칡을 대상으로 캘러스의 대량증식을 통한 이퀄의 전구물질인 이소플라본의 대량생산에 기여할 수 있으며, 기내에서 생물전환을 통하여 이퀄을 생산하기 위한 기반이 될 것이다.

• 대한한의학회지
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• 제31권6호
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• pp.8-15
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• 2010

Objectives: We investigated to develop and validate HPLC-PDA methods for simultaneous determination of eight constituents in Galgeun-tang (GGT). Methods: Reverse-phase chromatography using a Gemini C18 column operating at $40^{\circ}C$, and photodiode array (PDA) detection at 230 nm, 254 nm, and 280 nm, were used for quantification of the eight marker components of GGT. The mobile phase using a gradient flow consisted of two solvent systems. Solvent A was 1.0% (v/v) aqueous acetic acid and solvent B was acetonitrile with 1.0% (v/v) acetic acid. Results: Calibration curves were acquired with $r^2$ > 0.9999, and the relative standard deviation (RSD) values (%) for intra- and inter-day precision were less than 3.0%. The recovery rate of each component was in the range of 87.33-101.38%, with an RSD less than 7.0%. The contents of the eight components in GGT were 1.98-12.17 mg/g. Conclusions: The established method will be applied for the quantification of marker components in GGT.

• 생약학회지
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• 제40권3호
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• pp.224-228
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• 2009

For the quality control of traditional herbal medicine, Galgeun tang, simultaneous determination of glycyrrhizin, paeoniflorin, puerarin, 6-gingerol was established by using a high performance liquid chromatographic (HPLC) method with diode array detector. To separate five four constituents, DIONEX $C_{18}$ column ($5{\mu}m$, $120{\AA}$, $4.6\;mm{\times}150\;mm$) was used with gradient elution system of water and methanol. Validation of the chromatography method was evaluated by linearity, recovery, and precision test. Calibration curve of standard components showed excellent linearity ($R^2$>0.9906). Limits of detection (LOD) and limits of quantification (LOQ) varied from 0.15 to $0.52{\mu}g$/ml and 0.27 to $0.80{\mu}mg$/ml, respectively. The relative standard deviations (RSDs) of data of the intra-day and inter-day experiments were less than 2.88% and 1.21%, respectively. The results of recovery test were ranged from 96.71 to 106.29% with RSD values 0.01-0.80%.

• Natural Product Sciences
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• 제7권3호
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• pp.63-67
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• 2001

${\beta}-Glucuronidase-inhibitory$ and hepatoprotective effects of Chungpesagan-tang, which has been used for liver diseases and stroke, on $CCl_4-induced$ hepatotoxicity of rats was investigated. Chungpesagan-tang potently inhibited ${\beta}-glucuronidases$. Serum AST, ALT and LDH levels of the $CCl_4$ group orally treated with Chungpesagan-tang (100 mg/kg) were lowered to 47, 28 and 58% of the $CCl_4-treated$ group, respectively. Among the ingredients of Chungpesagan-tang, Puerariae Radix, Scutellariae Radix and Rhei Rhizoma potently inhibited the ${\beta}-glucuronidase$ and protected $CCl_4-induced$ liver injury. The hepatoprotective activity of Puerariae Radix was affected by ingredients of Chungpesagan-tang: Scutellariae Radix had the synergistic activity, but Angelicae Tenussimae Radix exhibited the antagonistic activity. These results suggest that the ${\beta}-glucuronidase$ inhibitor of herbal medicines may protect $CCl_4-induced$ liver injury and puerarin should be a natural prodrug for the hepatoprotective effect.

• Archives of Pharmacal Research
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• 제21권1호
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• pp.17-23
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• 1998

Flavonoid glycosides were metabolized to phenolic acids via aglycones by human intestinal microflora producing ${\alpha}$-rhamnosidase, exo-${\beta}$-glucosidase, endo- ${\beta}$-glucosidase and/or ${\beta}$-glucuronidase. Rutin, hesperidin, naringin and poncirin were transformed to their aglycones by the bacteria producing ${\alpha}$-rhamnosidase and ${\beta}$-glucosidase or endo- ${\beta}$-glucosidase, and baicatin, puerarin and daidzin were transformed to their aglycones by the bacteria producing ${\beta}$glucuronidase, C-glycosidase and ${\beta}$-glycosidase, respectively. Anti-platelet activity and cytotoxicity of the metabolites of flavonoid glycosides by human intestinal bacteria were more effective than those of the parental compounds. 3,4-Dihydroxyphenylacetic acid and 4-hydroxyl-phenylacetic acid were more effective than rutin and quercetin on anti-platelet aggregation activity. 2,4,6-Trihydroxybenzaidehyde, quercetin and ponciretin were more effective than rutin and ponciretin on the cytotoxicity for tumor cell lines. We insist that these flavonoid glycosides should be natural prodrugs.

• 생약학회지
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• 제40권1호
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• pp.46-50
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• 2009

The antitumor activity of the roots extract of Pueraria thunbergiana was investigated on the basis of cytotoxicity upon the cultured human tumor cell lines, in vitro. The purification of methylene chloride (MC) soluble part and ethylacetate (EA) soluble part of extract by column chromatography furnished seven isoflavonoids, two triterpenoids, one but-2-enolide. The structures of them were established by chemical and spectroscopic means to be lupeol (1), $\beta$-sitosterol (2), biochanin A (3), (-)-tuberosin (4), calycosin (5), daidzein (6), puerarin (7), daidzin (8), (+)-puerol-B 2-O-$\beta$-glucopyranoside (9), formononetin-7-O-$\beta$-glucopyranoside (10). Each isolates ($1{\sim}10$) were evaluated for inhibitory activities on the proliferation of cultured human tumor cell lines such as A549, SK-OV-3, HCT-15 and SK-MEL-2, respectively.

• Bulletin of the Korean Chemical Society
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• 제30권7호
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• pp.1459-1462
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• 2009

Two new flavone glycosides, giraldiin A and B, together with three known compounds, annulatin, myricetin 3-O-$\alpha$- L-rhamnopyranoside and gallic acid, were isolated from the ethanol extract of the root of Pteroxygonum giraldii Damm. et Diels. The structures of giraldiin A and B are designated as 3'-($\alpha$-L-arabinopyranosyloxy)-4',5,5',7- tetrahydroxy-3-methoxyflavone and 4'-($\beta$-D-glucopyranosyloxy)-5,5',7-trihydroxy-2',3-dimethoxyflavone, respectively, on the basis of detailed spectroscopic analyses. The free radical scavenging activity of giraldiin A was evaluated by decolouring spectrophotometry of pentamethine cyanine dye (Cy5) with $Fe^{2+}-H_2O_2$ Fenton radical generating system. The results indicated the hydroxyl free radical scavenging activity of giraldiin A (E$D_{50}$ = 23.7 nmol/mL) is higher than that of some known antioxidants such as rutin, puerarin, daidzein and 2,6-di-tertbutyl-4-methylphenol.

• Archives of Pharmacal Research
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• 제25권2호
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• pp.165-169
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• 2002

The herbal components should be transformed to bioactive compounds by the intestinal bacteria and then expressed the pharmacological action of herbal medicines. Human fecal enzyme activities related to the metabolism of herbal components were measured. The metabolic activities of puerarin, poncirin, glycyrrhizin, ginsenoside Rb1 and ginsenoside Rb2 to their bioactive compounds were $3.5{\pm}1.18,{\;}333.1{\pm}183.64,{\;}95.7{\pm}107.1,{\;}20.8{\pm}10.32{\;}and{\;}20.8{\pm}13.3{\;}{\mu}mo1/h/g$, respectively. The profile of these metabolic activities of glycyrrhizin and ginsenosides were not changed even if herbal extracts, water extract of Glycyrrhizae Radix and Ginseng Radix, instead of the isolated compounds were used . All the enzyme activities tested were not different between male and female, and between ages. However, the difference of these enzyme activities in individuals was significant. These results suggest that the metabolic activity of herbal components to bioactive compounds may be a factor of constitutional classification, and could be available for constitutional classifications, if the constitutional herbal medicines were used .

• 동의생리병리학회지
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• 제33권1호
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• pp.39-47
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• 2019

Galgeun-tang (GGT) is a traditional herbal medicine generally used to treat cold, fever, headache, migraine and convulsion. To investigate the difference of ingredient and efficacy of GGT according to production method, mix extract powder and decoction, the quantities of index components, antioxidant and anti-inflammatory effects of two kinds of GGT formulation were compared. The contents of ten index components were simultaneously analyzed with HPLC. A HPLC method was developed and validated for the simultaneous determination. The GGT mix extract powder contained higher puerarin and daidzin content than the decoction, while the GGT decoction contained higher cinnamic acid and glycyrrhizic acid content than the mix extract powder. But ingredient of GGT mix extract powder and decoction was statistically very similar (r=0.8335). GGT decoction showed higher antioxidant effect than GGT mix extract powder, but the difference was low. The patterns of productions for NO, iNOS, COX-2, TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 were decreased along the increasing dose of GGT formulation in LPS-induced RAW 264.7 macrophages. The inhibitory effects on NO, iNOS, COX-2, IL-$1{\beta}$ and IL-6 in Raw 264.7 cell were very similar to each other. The results obtained in this study could be used as fundamental data for verifying the equivalence and effectiveness of the herbal medicines.

• 생명과학회지
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• 제29권5호
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• pp.545-554
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• 2019

칡의 화합물은 동양 전통 의학에서 매우 중요한 콩과 식물로서 다량의 이소플라본을 함유한다고 보고되었다. 그러나, 칡의 주요 함유 이소플라본은 생물학적으로 이용가능성이 낮기 때문에, 생물학적 이용가능성을 증대시키기 위해서는 가수분해나 ${\beta}$-glucosidase를 사용하여 생물전환을 통해 이용 효율성을 증대시킬 수 있다. 본 연구는 김치로 부터 분리된 Lactobacillus rhamnosus BHN-76를 이용해 발효한 칡 추출물의 항산화 효과를 조사하였다. L. rhamnosus BHN-76 발효는 $37^{\circ}C$에서 3일간 발효하였으며, 발효하지 않은 칡 추출물에 비해 L. rhamnosus BHN-76 발효 칡 추출물에서 총 폴리페놀 함량은 약 134%, 총 플라보노이드 함량은 약 110% 증가된 것을 확인하였다. 또한, SOD 유사활성능, DPPH radical 소거 활성능과 ABTS radical 소거활성능은 각각 약 213%, 190%와 107% 항산화능이 증가하는 것을 확인하였다. 이 결과를 통해 L. rhamnosus BHN-76을 이용한 칡의 발효가 가능하며, 유산균 발효가 칡의 항산화능 증대에 효과적인 것을 확인하였으며 본 연구를 기반으로 한 기능성 식품 또는 화장품 소재로의 개발 및 응용이 가능할 것으로 기대된다.