Solvent-partitioned Fractions Research Articles

Cassia fistula leaves extract profiling and its emphasis on induced ulcerative colitis in male rats through inhibition of caspase 3 and cyclooxygenase-2

The objective of the study was to determine the total phenolic and total flavonoid contents (TPC and TFC, respectively), as well as the solvent-partitioned fractions, of the leaf extract of Cassia fistula. Using DPPH, TAC, and FRAP tests, the in vitro antioxidant properties of crude extract and its ethyl acetate fraction were investigated. Additionally, C. fistula chemical profiling was accomplished using LC-ESI-MS/MS. Along with that, the effectiveness of crude extract (200 and 400 mg/kg) in treating male rats with ulcerative colitis (UC) induced by acetic acid was assessed. According to the findings, the ethyl acetate fraction had the highest concentrations of TPC (12.36 1.46 mg GAE/100 mg) and TFC (5.12 0.64 mg QE/100 mg), as well as impressive in vitro antioxidant activity with IC50 values of DPPH (12.7 g/mL), FRAP (4.87 0.71 mM Fe+2/g), and TAC (55. Fifty-five chemicals, predominantly phenolics (catechins, flavonoids, anthraquinones, chalcones, and phenolic acids), were detected by the LC/MS/MS. The anti-UC effect of C. fistula was dose-dependent. The hematological parameters, liver biomarkers, oxidative stress, histopathology, and immunohistochemistry revealed that prophylactic administration of crude extract to colitis rats ameliorated all the previous biomarkers. However, when the crude extract was administered on established colitis, it facilitated the recovery of the inflamed mucosa and showed better results than the protection mode. The phytoconstituents of C. fistula that were discovered here by LC/MS/MS were examined by molecular docking studies for their binding affinities towards COX-2 and caspase-3 proteins to highlight the anti-inflammatory and antiapoptotic activities of the plant. The Emodin compound displayed the highest binding affinity for COX-2 proteins, while isorhamnetin was discovered to have the best binding associations with the essential amino acids of caspase-3. Procyanidin B2 interestingly shows potent interactions with important amino acids of two targets. This study has made it possible to utilize C. fistula in the treatment of UC and has clarified its mechanism of action.

Kinetics of trypsin inhibition by methanolic and solvent-partitioned fractions of two medicinal plants – Momordica charantia and Xylopia aethiopica

Kinetics of trypsin inhibition by methanolic and solvent-partitioned fractions of two medicinal plants – Momordica charantia and Xylopia aethiopica

Evaluation and Comparison of the Antimelanogenic Properties of Different Solvent Fractionated Cnidium japonicum Extracts in B16F10 Murine Melanoma Cells.

Cnidium japonicum is a biennial halophyte found in the salt marshes and shores of Korea and widely used in traditional Korean medicine as an ingredient. This study investigated and compared the antimelanogenic effect of solventpartitioned fractions of C. japonicum extract (CJEFs) in a B16F10 mouse melanoma cell model, focusing on tyrosinase activity and production. Melanogenesis is the process in which skin pigment melanin is produced through tyrosinase activity. Overproduction of melanin is the primary reason behind several skin disorders such as freckles, spots, and hyperpigmentation. The antimelanogenic capacity of CJEFs was initially screened by their tyrosinase inhibitory effects, prevention of dihydroxyphenylalanine (DOPA) oxidation, and suppression of melanin production. The inhibition of tyrosinase activity and DOPA oxidation by CJEFs was suggested to be related to the downregulation of microphthalmia-associated transcription factor, tyrosinase, tyrosinase-related protein-1, and tyrosinase-related protein-2, which was confirmed using mRNA and protein expression levels. Moreover, the glycogen synthase kinase 3 beta- and cyclic adenosine monophosphate response element-binding protein-related signaling pathways were inhibited by treatment with CJEFs, indicating their action mechanism. All the tested CJEFs exerted similar effects on tyrosinase activity and production. However, among those, 85% aq. MeOH was the most active fraction to suppress the signaling pathway that produces tyrosinase. These results suggest that especially the MeOH fraction of C. japonicum extract serves as a potential source of bioactive substances, with effective antimelanogenesis properties.

ACE Inhibitory Activity and Functional Group Analysis of Solvent- Partitioned Fractions of Eleusine indica

Eleusine indica Linn. Gaerth or Indian goose grass has been widely used as an alternative natural anti-hypertensive agent. However, the rationale behind this mechanism is still unknown. The purpose of this study is to confirm the presence of functional groups that may be responsible for its ACE inhibitory activity and its possible synergism. Each solvent-portioned fraction was analyzed by FTIR, individual ACE inhibitory activity, and combination of fractions. FTIR results of solvent- partitioned fractions revealed that O-H and C-H stretches were present in all fractions. In addition to this, C-O and C=O groups were also present in all fractions except in ethyl acetate. S=O groups were also present in chloroform, ethyl acetate, and n-butanol fractions. Other function groups such as N-O and C=C groups were present in ethyl acetate and n-butanol fractions, respectively. The highest % inhibition obtained among the individual fraction is n-butanol at 96.031 % ± 0.004 at 500 ug/ml. The lowest % inhibition obtained among the individual fraction is chloroform at 32.544 % ± 0.011 at 500 ug/ml. The highest % inhibition among the combined fractions is ethyl acetate:butanol at 95.727% ± 20.847, while the lowest % inhibition in combination is hexane:chloroform at 89.626 % ± 19.577. The fraction with the lowest IC50 is hexane at 15.357, while the highest IC50 is chloroform at 168.207 ug/ml. Functional group present may be associated with the antihypertensive activity of the E. indica extracts. The more polar solvent-partitioned fractions of E. indica has a higher ACE inhibitory activity compared to its less polar solvent- partitioned fractions. The higher ACE inhibitory activity may be associated with the secondary metabolites present in the more polar fractions. There is also possible synergism present in the combined solvent-partitioned fractions as their % inhibition is higher than their individual effects.

Chemical Characterization and Antioxidant Properties of Ethanolic Extract and Its Fractions from Sweet Potato (Ipomoea batatas L.) Leaves.

Sweet potato (Ipomoea batatas L.) leaf is a natural source of phenolic compounds with strong antioxidant activity and potential utility as an antioxidant. The aim of this study was to evaluate the polyphenol composition and antioxidant activities of ethanol extracts and their various solvent-partitioned fractions (petroleum ether, ethyl acetate, and aqueous fraction) from sweet potato leaves and petioles. Seven caffeoylquinic acid (CQA) derivatives and four flavonoids were detected in sweet potato leaves by HPLC-ESI-MS. The total phenolic content (TPC) and total flavonoid content (TFC) in leaf (112.98 ± 4.14 mg gallic acid equivalent (GAE)/g of dried extract, 56.87 ± 5.69 mg rutin equivalent (RE)/g of dried extract) was more than ten times higher than in petiole (9.22 ± 2.67 mg GAE/g of dried extract, 3.81 ± 0.52 mg RE/g of dried extract). The antioxidant contents of ethyl acetate fractions increased dramatically relative to those of crude extracts for both leaves and petioles. Purification using solvent partition with ethyl acetate increased TPC and TFC of crude extracts, especially the CQA derivatives including 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, and 3,4,5-tricaffeoylquinic acid. Meanwhile, the ethyl acetate fractions with the highest CQA content were associated with the highest scavenging activities towards 2,2-diphenyl-1-picrylhydrazyl (DPPH) and higher ferric ion reducing antioxidant power (FRAP)-reducing power.

Anti-photoaging effects of solvent-partitioned fractions from Portulaca oleracea L. on UVB-stressed human keratinocytes.

In this study, the anti-photoaging activity of solvent-fractionated extracts from Portulaca oleracea L. was investigated in ultraviolet B-stressed keratinocytes. The results showed that the treatment of the fractions suppressed the generation of intracellular reactive oxygen species (ROS) and activated mRNA and protein expression levels of superoxide dismutase1 (SOD-1) and heme oxygenase1 (HO-1) enzymes via Nrf-2 pathway. The tested fractions down-regulated the expression of matrix metalloproteinases (MMPs) and up-regulated type Ⅰ procollagen production. Among the tested samples, the 85% aq. MeOH fraction was the most effective in suppressing MMPs and type Ⅰ procollagen degradation. Two homoisoflavonoids, portulacanone A (PA) and portulacanon D (PD), were isolated from the 85% aq. MeOH fraction. PA and PD successfully inhibited the secretionof MMP-1 and increased the production of type Ⅰ procollagen in UVB-stressed keratinocytes. Therefore, solvent-partitioned factions from P. oleracea and its active components, PA and PD, were suggested to have beneficial effects against photoaging. PRACTICAL APPLICATIONS: Portulaca oleracea L. is an edible nutrient-rich herb with high amounts of omega-3 and omega-6 fatty acids, ascorbic acid, and dietary minerals. P. oleracea has also been used as a botanical medicine for its potential health benefits such as antioxidant, antibacterial, and antiinflammatory properties. The current study suggested that the extracts from P. oleracea were able to prevent human keratinocytes from UVB-induced stress and damage. Due to its protective abilities, it can be used as an additive in the cosmeceutical industry to produce novel skin protectants. In addition, due to its intracellular antioxidant and MMP inhibitory effects, it can be a valuable source for nutraceutical development. Further studies that characterize the isolated flavonoids may also yield bioactive molecules to be utilized in cosmeceutical and nutraceutical industries as lead compounds. Nevertheless, P. oleracea can serve as a health beneficial addition to topical applications and dietary supplements against UVB-stimulated oxidative stress in skin cells.

Inhibitory Effects of Solvent-Partitioned Fractions of Two Nigerian Herbs (Spondias mombin Linn. and Mangifera indica L.) on α-Amylase and α-Glucosidase.

Therapies directed towards controlling hyperglycemia, the hallmark of type-2 diabetes mellitus, go a long way in managing diabetes and its related complications. Reducing glucose level through the inhibition of the relevant carbohydrate hydrolyzing enzymes is one among many routes in the management of diabetes. This study investigates the in vitro enzyme inhibitory and antioxidant properties of solvent-partitioned fractions of Spondias mombin and Mangifera indica leaves; which are used extensively in the treatment of diabetic patients locally. The leaves of S. mombin and M. indica were extracted with methanol and fractionated to obtain n-hexane (HF), ethyl acetate (EAF), n-butanol (BF), and aqueous (AF) fractions successively. The α-amylase and α-glucosidase inhibitory activities of fractions of S. mombin and M. indica leaves were investigated while the antioxidant activity of each fraction was analyzed using iron chelating and ABTS (2,2’-azino-bis(3-ethylbenzothiazoline)-6-sulphonic acid) radical scavenging assay. Our findings indicated that the ethyl acetate fraction of M. indica leaves contained a considerably higher (p < 0.05) amount of total phenolic, flavonoids, metal ion, and ABTS radical scavenging activity than the ethyl acetate fractions of S. mombin. Furthermore, the ethyl acetate fraction of M. indica had a considerably higher (p < 0.05) inhibitory effect on α-glucosidase (IC50 = 25.11 ± 0.01 μg mL−1), and α-amylase (IC50 = 24.04 ± 0.12 μg mL−1) activities than the S. mombin fraction. Hence, the inhibitory activities of S. mombin and M. indica leaves suggest that they are a potential source of orally active antidiabetic agents and could be employed to formulate new plant-based pharmaceutical and nutraceutical drugs to improve human health.

Solvent-partitioned fractions from Ishige okamurae extract inhibit MMP-2 and MMP-9 activities in human fibrosarcoma cells in vitro

Matrix metalloproteinases (MMPs) are endopeptidases that take significant roles in extracellular matrix degradation and therefore linked with several complications such as metastasis of cancer progression, oxidative stress, and hepatic fibrosis. Ishige okamurae, a brown alga, has been reported to possess various bioactivities including but not limited to antiviral, antimicrobial, and anti-inflammatory. In this study, the potential of I. okamurae against cancer cell invasion was evaluated through MMP inhibitory effect in HT1080 fibrosarcoma cells in vitro. I. okamurae crude extract was fractionated with organic solvents, namely water (H2O), n-butanol (n-BuOH), 85% aqueous methanol (85% aq. MeOH), and n-hexane (n-Hex). The non-toxicity of fractions was confirmed by MTT assay. All fractions inhibited the enzymatic activities of MMP-2 and MMP-9 according to gelatin zymography assay. Cell migration was also inhibited by 85% aq. MeOH fraction, significantly. In addition, both gene and protein expressions of MMP-2 and MMP-9, and tissue inhibitor of MMPs (TIMPs) were evaluated by RT-PCR and Western blotting, respectively. Fractions suppressed the mRNA and protein levels of MMP-2 and MMP-9 while elevating the TIMP-1 and TIMP-2, H2O fraction being the least effective while 85% aq. MeOH fraction the most. Collectively, 85% aq. MeOH fraction from brown algae could be a potential inhibitor of MMPs, suggestively considering presence of poly-unsaturated fatty acids in high amounts.

염생식물 가는갯는쟁이 용매 추출물의 항염증활성

본 연구에서는 염생식물을 이용한 생리활성 소재 개발 연구의 일환으로 가는갯능쟁이의 용매 추출물 및 분획물을 제조하여 LPS로 염증을 유도한 마우스 유래 macrophages에서의 NO 생성 및 염증관련인자의 발현에 미치는 영향을 검토하여 가는갯능쟁이의 항염증 소재로서의 이용 가능성을 검토하였다. 가는갯능쟁이는 methylene chloride와 methanol을 이용하여 제조한 조추출물(crude extract, 48.3 mg)을 용매극성에 따라 단계적으로 분획하여 n-hexane 층(11.5 mg), 85% aqueous methanol 수용액층(85% aq. MeOH, 11.3 mg), n-butanol 층(n-BuOH, 11.2 mg)과 water 층(12.3 mg)을 얻었다. 가는갯능쟁이의 조추출물 및 분획물의 RAW264.7 대식세포에 대한 독성 여부를 측정하여 세포에 대한 독성이 나타나지 않는 처리 농도에서 추출물 및 분획물의 항염증 활성을 확인하였다. 조추출물은 LPS로 유도된 NO 생성을 유의적으로 억제하는 효과를 나타내었으며, 주요 염증 유발인자인 iNOS와 COX-2의 발현 또한 효과적으로 억제하였다. 분획물의 NO 생성 억제효과는 85% aq. MeOH> n-BuOH >n-hexane > <TEX>$H_2O$</TEX>의 순으로 나타났으며, 세포내 전사수준에서 염증관련 유전자의 발현 억제율은 <TEX>$H_2O$</TEX> 분획 처리군에 비해 85% aq. MeOH, n-BuOH 및 n-hexane 분획 처리군에서 높게 나타났다. 이상의 결과로부터 항염증 활성이 가장 높게 나타난 85% aq. MeOH 분획층으로 부터 염증억제 효과가 있는 유효 성분의 분리가 기대된다. As a part of ongoing research to elucidate and characterize antiinflammatory nutraceuticals, the crude extracts from Atriplex gmelinii C. A. Mey. and their solvent-partitioned fractions were tested for their antiinflammatory potential in lipopolysaccharide (LPS)-stimulated RAW 264.7 mouse macrophages. The crude extracts of A. gmelinii C. A. Mey. were fractioned according to polarity with n-hexane, 85% aqueous methanol (85% aq. MeOH), n-butanol, and <TEX>$H_2O$</TEX>. Their antiinflammatory activities were investigated in LPS-induced inflammation in mouse macrophages by measuring nitric oxide (NO) generation and mRNA expression of inflammation mediators, namely, inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-<TEX>$1{\beta}$</TEX> (<TEX>$IL-1{\beta}$</TEX>), and IL-6. As a result, we confirmed that the crude extracts of A. gmelinii C. A. Mey. inhibited LPS-stimulated NO production and mRNA expression of iNOS and COX-2 as important inflammatory factors. The inhibition of NO production through the downregulation of important inflammatory factors such as iNOS, COX-2, <TEX>$IL-1{\beta}$</TEX>, and IL-6 was found by treatment with all solvent-partitioned fractions. Among all tested fractions, 85% aq. MeOH showed the strongest antiinflammatory response. Based on the current results, A. gmelinii C. A. Mey. was suggested to possess natural antiinflammatory components, indicating that it could be used as a valuable source of antiinflammatory substances.

Crude extract and solvent fractions of Calystegia soldanella induce G1 and Sphase arrest of the cell cycle in HepG2 cells.

The representative halophyte Calystegia soldanella (L) Roem. et Schult is a perennial vine herb that grows in coastal dunes throughout South Korea as well as in other regions around the world. This plant has long been used as an edible and medicinal herb to cure rheumatic arthritis, sore throat, dropsy, and scurvy. Some studies have also shown that this plant species exhibits various biological activities. However, there are few studies on cytotoxicity induced by C. soldanella treatment in HepG2 human hepatocellular carcinoma cells. In this study, we investigated the viability of HepG2 cells following treatment with crude extracts and four solvent-partitioned fractions of C. soldanella. Of the crude extract and four solvent fractions tested, treatment with the 85% aqueous methanol (aq. MeOH) fraction resulted in the greatest inhibition of HepG2 cell proliferation. Flow cytometry showed that the 85% aq. MeOH fraction induced a G0/G1 and S phase arrest of the cell cycle progression. The 85% aq. MeOH fraction arrested HepG2 cells at the G0/G1 phase in a concentration-dependent manner, and resulted in decreased expression of cyclin D1, cyclin E, cyclin-dependent kinase (CDK)2, CDK4, CDK6, p21, and p27. Additionally, the 85% aq. MeOH fraction treatment also arrested HepG2 cells in the S phase, with decreased expression of cyclin A, CDK2, and CDC25A. Also, treatment with this fraction reduced the expression of retinoblastoma (RB) protein and the transcription factor E2F. These results suggest that the 85% aq. MeOH fraction exhibits potential anticancer activity in HepG2 cells by inducing G0/G1 and S phase arrest of the cell cycle.

Anti-Inflammatory Activity of the Solvent-Partitioned Fractions from Spergularia marina in LPS-Stimulated RAW 264.7 Cells.

As a part of ongoing research to elucidate and characterize antioxidant and anti-inflammatory nutraceuticals, solvent-partitioned fractions from Spergularia marina were tested for their ability to scavenge radicals and suppress inflammation. The results of the 2′,7′-dichlorofluorescein diacetate assay indicate that solvent-partitioned fractions from S. marina scavenged intracellular radicals in H2O2-stimulated mouse macrophages. The tested fractions decreased the generation of nitric oxide (NO) and the expression of inflammation mediators, namely, inducible nitric oxide synthase (iNOS) and interleukin (IL)-6, by lipopolysaccharide (LPS)-induced mouse macrophages, indicating that S. marina decreases inflammation. Among all tested fractions [i.e., H2O, n-buthanol (n-BuOH), 85% aqueous methanol (aq. MeOH), and n-hexane], the 85% aq. MeOH fraction showed the strongest antioxidant and anti-inflammatory response. The 85% aq. MeOH fraction scavenged 80% of the free radicals produced by H2O2-induced control cells. In addition, NO production was 98% lower in 85% aq. MeOH fraction-treated cells compared to LPS-induced control cells. The mRNA expression of iNOS and IL-6 was also suppressed in 85% aq. MeOH fraction-treated cells. The results of the current study suggest that the phenolic compound components of S. marina are responsible for its antioxidant and anti-inflammatory effects.

애기거머리말의 항산화 활성

In this study, crude extract of the seagrass Zostera japonica, and its solvent-partitioned fractions were evaluated for their antioxidant activity. The crude extract was successively fractionated into n-hexane, 85% aqueous methanol (85% aq.MeOH), n-butanol (n-BuOH), and water fractions by liquid-liquid partition. These include DPPH radical scavenging, hydroxyl radical scavenging in HT-1080 cells, peroxynitrite scavenging, and protective effect on DNA damage caused by hydroxyl radicals generated. In all assays, except for DPPH radical, 85% aq.MeOH and n-BuOH fraction showed the strong antioxidant activity. These results suggest that Z. japonica may be used as a potential source of natural antioxidants for the development of cosmetic product or functional food in the future.

Flavonoid content, free radical scavenging and increase in xanthine oxidase inhibitory activity in Galgeun-tang following fermentation with Lactobacillus plantarum.

Galgeun-tang (GT) prior to and following fermentation with Lactobacillusplantarum was analyzed to determine the total polyphenol and flavonoid contents and the antioxidant activity. GT, fermented GT (FGT) and their three solvent-partitioned fractions, which were prepared by successive partitioning with ethyl acetate (EtOAc), butanol (BuOH) and water, were evaluated for total polyphenol and flavonoid contents, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and xanthine oxidase (XO) inhibitory activity. Following fermentation, the total polyphenol content only increased slightly; however, the flavonoid content increased by 24.3%. The radical scavenging activity increased from 22.4 to 27.5% and the XO inhibitory activity increased from 20.2 to 62.4% at 500µg/ml. The EtOAc fraction among the solvent‑partitioned fractions demonstrated the highest total polyphenol and flavonoid contents, radical scavenging activities and XO inhibitory activity, and the quantity also markedly increased following fermentation.

Antipsychotic property of solvent-partitioned fractions of Lonchocarpus cyanescens leaf extract in mice

This study was carried out to evaluate the antipsychotic property of solvent-partitioned fractions of the leaf extract of Lonchocarpus cyanescens (LC), a reputable medicinal plant used in folk medicine for the treatment of mental illnesses in Nigeria. The n-hexane fraction, ethyl acetate fraction (EAF), and aqueous ethanol fraction (AEF) of LC were tested for antipsychotic property based on the antagonism of stereotypy induced by apomorphine (APO). Antagonism of hyperactivity and lethality in aggregated mice induced by amphetamine (AMPH) were further employed for screening the antipsychotic effect of the fractions. EAF was tested for catalepsy utilising the horizontal plane paradigm. Thin-layer chromatography (TLC) was used to screen EAF for the presence of secondary metabolites. AEF (100-400 mg/kg) significantly (p<0.05) suppressed stereotypy induced by APO (2 mg/kg, intraperitoneal [IP]) in comparison with control, suggesting antipsychotic activity. However, EAF (200 mg/kg, IP) was most potent in inhibiting the stereotypic effect of APO. EAF was also the most active in antagonising AMPH-induced hyperactivity and in protecting against death caused by AMPH in grouped mice. However, in contrast to haloperidol, EAF did not produce cataleptic behaviour in the horizontal plane paradigm. The TLC analysis revealed that EAF contains several compounds, with some of them having Rf values similar to that of haloperidol, which suggests the presence of active substances with the same chemical structural identity. These findings suggest that EAF contains the major active constituent(s) mediating the antipsychotic property of LC and further support its use for the management of psychosis in traditional medicine.

Evaluation of inhibitory effect of phlorotannins from Ecklonia cava on triglyceride accumulation in adipocyte.

In the present study, a methanolic extract of Ecklonia cava and its solvent-partitioned fractions were evaluated for their antiadipogenic effect in 3T3-L1 adipocytes. One of them, the n-BuOH fraction, effectively reduced lipid accumulation and glucose consumption. In addition, the presence of the n-BuOH fraction in adipocytes suppressed the regulations of adipogenic transcription factors, PPARγ and SREBP1c, and adipogenic specific genes, FABP4, FABP1, FAS, LPL, HSL, and ACS1. Further purification of n-BuOH fractions led to the isolation of six phlorotannins (1-6). The six phlorotannins effectively suppressed triglyceride accumulation. Comparative analysis showed that lipid accumulation in adipocytes was dramatically attenuated in the presence of eckstolonol (4).

애기거머리말 추출물의 암세포 성장 억제효과

In this study, crude extracts of the marine eelgrass Zostera japonica and their solvent-partitioned fractions were evaluated for their inhibitory effect against AGS, HT-1080 and MCF-7 human cancer cells using MTT assay. Each of the crude extracts (acetone/methylene, chloride, and methanol) of Z. japonica showed a significant inhibitory effect on the growth of human cancer cells. The combined crude extracts were partitioned between CH₂Cl₂ and water. The organic layer was further partitioned between 85% aq. MeOH and n-hexane, and the aqueous layer was then fractionated into n-BuOH and H₂O, successively. Growth inhibition effects of solvent-partitioned fractions from Z. japonica on human cancer cells increased in a dose-dependent manner. Among these tested samples, the 85% aq. MeOH fraction revealed good inhibitory effects on the growth of AGS and HT-1080 human cancer cells, while the n-hexane fraction exhibited good inhibitory effects on the growth of AGS and MCF-7 human cancer cells. In addition, 85% aq. MeOH and n-hexane fractions enhanced mRNA expression of p53 gene. These results suggest that there is further scope for the isolation of active compounds from Z. japonica, which should show much stronger anticancer activity.

참모자반 메탄올 추출 분획물의 항산화 및 숙취해소능과 α-glucosidase 활성저해효과

참모자반 분획물은 극성에 따라 핵산, 메탄올, 부탄올 및 수층 순으로 분획하여 4개의 분획층을 얻었으며, 각 분획층의 생리활성을 요약하면 다음과 같다. 항산화 활성은 DPPH radical 소거능과 SOD 활성을 측정한 결과 모두 농도 의존적으로 증가하였으며 특히 메탄올 분획 층에서 가장 높은 항산화 효과를 보였다. 아질산염 소거능 실험에서는 부탄올 층이 pH 1.2에서 57.8%로 4개의 분획물중 가장 높은 아질산염 소거능을 나타냈다. 숙취 해소능을 측정하기 위하여 ADH 활성과 ALDH 활성을 측정하였다. 대조구로 사용한 흡광도 값을 100으로 하였을 때 positive control로 사용한 hepos가 150~167%의 활성을 나타내었으며 4가지 분획층 모두 농도 의존적으로 활성이 증가하였다. 특히 ALDH에서는 메탄올 분획층이 10 mg/ml 농도에서 167.4%의 활성을 나타내어 hepos와 비슷한 효과를 보였다. 참모자반 분획물의 혈당강하 효과는 <TEX>${\alpha}$</TEX>-glucosidase 활성억제 효과로 측정하였다. <TEX>${\alpha}$</TEX>-glucosidase 활성억제 효과는 핵산 분획층에서 2 mg/ml의 농도에서 94.1%로 아주 높은 항당뇨 효과를 나타내었다. 그런데 부탄올층, 메탄올층 및 수층은 혈당강하효과가 거의 없는 것으로 나타났다. 피부주름예방을 측정하기 위하여 수행한 elastase 저해효과 실험에서 메탄올 분획층에서 농도 의존적으로 높은 주름개선효과가 나타났으며 2 mg/ml 농도에서 93.2%의 효과를 나타냈으며, positive control로 사용한 Vit. C (1 mg/ml) 보다 더 높은 활성을 나타냈다. 이러한 결과는 모자반 메탄올 추출물 중 특히 메탄올 분획층이 탁월한 주름개선 효과, 숙취 해소능과 항산화 효과가 있는 것으로 사료되며, 핵산층은 혈당강하 효과가 높은 것으로 나타났다. 따라서 모자반 유기용매 분획물은 천연 기능성소재로서의 사용 가능성이 매우 높을 것으로 사료된다. We investigated the physiological activity and solvent-partitioned fractions of methanol extracts from the green seaweed Sargassum fulvellum. The methanol extract from S. fulvellum was sequentially fractionated with n-hexane (SFMH), methanol (SFMM), buthanol (SFMB), and water (SFMA). We investigated the antioxidant activities of solvent fractions from S. fulvellum by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging capacity and an SOD activity assay. DPPH radical scavenging capacity of SFMM was 79.5% at 10 mg/ml. SOD activity of SFMM was 79.9% at 10 mg/ml. Nitrite scavenging activities of solvent fractions from S. fulvellum were investigated under different pH conditions and showed the most remarkable effect at pH 1.2. In particular, the activity of SFMB was higher than the other fractions. ADH activity and ALDH activity of SFMM were 177.0% and 167.4% at 10 mg/ml, respectively. <TEX>${\alpha}$</TEX>-Glucosidase inhibitory activity of SFMH increased in a dose-dependent manner and was about 94.1% at 2 mg/ml. Elastase inhibitory activity was 93.2% at 2 mg/ml. These results revealed that S. fulvellum extracts have strong antioxidant and alcohol dehydrogenase activities and <TEX>${\alpha}$</TEX>-glucosidase inhibitory activity, suggesting that S. fulvellum extracts have potential as a source of natural products for health and beauty.

Evaluation on anti-adipogenic activity of flavonoid glucopyranosides from Salicornia herbacea

In the present study, the bioactivity-monitored isolation of flavonoid glucopyranosides from the salt-marsh plant Salicornia herbacea was performed, and their effects on adipogenic differentiation were evaluated in 3T3-L1 adipocytes. Among the solvent-partitioned fractions from S. herbacea, the 85% aq. MeOH and n-BuOH fractions effectively reduced the levels of triglyceride accumulation, glucose consumption and adipogenic transcription factor peroxisome proliferator-activated receptor-γ (PPARγ) expression. Further purification of the n-BuOH fraction led to the isolation of two flavonoid glucopyranosides, isorhamnetin 3-O-β-d-glucopyranoside (A) and quercetin 3-O-β-d-glucopyranoside (B), as the active principles. The presence of glucopyranosides A and B in adipocyte cells effectively suppressed adipogenic differentiation. Comparative analysis indicated that adipogenic differentiation was dramatically attenuated in the presence of glucopyranoside B rather than glucopyranoside A. The anti-adipogenic activity of glucopyranosides B was confirmed by down-regulation of sterol regulatory element-binding protein 1 (SREBP1), CCAAT/enhancer-binding proteins (C/EBPα), PPARγ and the adipocyte-specific proteins. Moreover, the specific mechanism mediating the effects of glucopyranosides B was confirmed by activation of AMP-activated protein kinase (AMPK). Findings from this study should underscore the nutraceutical value of S. herbacea-derived glucopyranosides as potent anti-obesity agents via alleviation of lipid accumulation.

菹草提取物对3种赤潮微藻生长的抑制作用&amp;lt;br&amp;gt;Growth Inhibition of the Three Species of Red Tide Microalgae by Extracts from Potamogeton crispus

以甲醇浸泡菹草干粉4 d，过滤、减压蒸干后，制备到菹草甲醇浸膏。此浸膏用90%甲醇水溶液(V/V)溶解后，依次以石油醚、乙酸乙酯和正丁醇为溶剂，采用液液萃取法进一步分离。经过滤、减压蒸干，获得4种液液分离提取物-石油醚、乙酸乙酯、正丁醇和蒸馏水提取物，得率分别为38.5%、21.9%、9.37%和26.6%。在此基础上，通过测定藻细胞数量，观察藻细胞形态，分析藻细胞内叶绿素、蛋白质和多糖含量等生理指标的变化，研究了此4种液液分离提取物对米氏凯伦藻(Karenia mikimitoi)、中肋骨条藻(Skeletonema costatum)和塔玛亚历山大藻(Alexandrium tamarense)生长的影响。结果表明，石油醚和乙酸乙酯提取物能明显降低3种赤潮微藻的细胞数量。当提取物浓度为16 mg/L时，此2种提取物对3种赤潮微藻的生长抑制率超过66%(第12 d)。同时，此2种提取物显著改变了3种赤潮微藻的细胞形态，致使藻细胞出现空洞、破碎和色素明显减褪现象。并且，它们还能显著影响藻细胞内叶绿素、蛋白质和多糖等生理指标的合成。在实验设定浓度范围内，正丁醇和蒸馏水提取物未对3种赤潮微藻的生长产生明显的影响。 To study effects of solvent-partitioned fractions of methanol extracts from Potamogeton crispus on the growth of the three species of microalgae (Karenia mikimitoi, Skeletonema costatum and Alexandrium tamarense), the extracts were extracted with petroleum ether, ethyl acetate, n-butanol and distilled water by liquid-liquid fractionaction, respectively. And yields of solvent-partitioned fractions were 38.5% petroleum ether extracts, 21.9% ethyl acetate extracts, 9.37% n-butanol extracts and 26.6% distilled water extracts, respectively. Based on the observation of algal morphology and the measurement of algal number, and the contents of physiological indicators (protein, polysaccharide and chlorophyll), the results showed the petro- leum ether extracts and ethyl acetate extracts from methanol extracts of Potamogeton crispus had the stron- gest action. The inhibitory effect of the tested microalgae by these two extracts was above 66% in day 12 at the concentration of 16 mg/L. And the petroleum ether extracts and ethyl acetate extracts caused cavities, pieces and chlorophyll decreased in cells. The further investigation found that these two extracts significantly decreased the contents of protein, polysaccharide and chlorophyll in the cells of those microalgae. But n- butanol extracts and distilled water extracts had not significant affect the growth of the all test microalgae.

Antioxidant activity of the halophyte Limonium tetragonum and its major active components

In this study, the antioxidant potentials of crude extracts and solvent-partitioned fractions of Limonium tetragonum were assessed by measuring their ability to scavenge intracellular reactive oxygen species (ROS) generated in HT-1080 cells. Following activity-oriented separation, four flavonol glycosides were isolated as active principles and their chemical structures were determined by 2 D NMR and by comparison with reported spectral data. The isolated compounds (1–4) were evaluated for their antioxidant capacity using three different activity tests; degree of occurrence of intracellular ROS, lipid peroxidation in HT-1080 cells and the extent of oxidative damage of genomic DNA purified from HT-1080 cells. All compounds exhibited significantly inhibited the generation of intracellular ROS and lipid peroxidation in HT-1080 cells, and significantly inhibited DNA oxidation. In addition, direct free radical scavenging effects of these compounds were investigated using the electron spin resonance (ESR) spin-trap technique.