• Title/Summary/Keyword: methanol extract

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Inhibition of Low Density Lipoprotein-oxidation, ACAT-1, and ACAT-2 by Lignans from the Bark of Machilus thunbergii

  • Shrestha, Sabina;Park, Ji-Hae;Lee, Dae-Young;Cho, Jin-Gyeong;Lee, Do-Gyeong;Cho, Moon-Hee;Jeong, Tae-Sook;Kang, Hee-Cheol;Baek, Nam-In
    • Journal of Applied Biological Chemistry
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    • v.54 no.1
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    • pp.63-66
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    • 2011
  • The bark of Machilus thunbergii was extracted with 80% aqueous methanol (MeOH), and the concentrated extract was partitioned using ethyl acetate (EtOAc), butanol (n-BuOH), and $H_2O$, successively. From the EtOAc fraction, five lignans were isolated through the repeated silica gel, octadecyl silica gel (ODS) and, Sephadex LH-20 column chromatography. Based on nuclear magnetic resonance (NMR), mass spectroscopy (MS), and infrared spectroscopy (IR) spectroscopic data, the chemical structures of the compounds were determined to be machilin A (1), machilin F (2), licarin A (3), nectandrin A (4), and nectandrin B, (5). This study presents comparative account of five lignans from M. thunbergii bark contributing inhibition of low density lipoprotein (LDL), ACAT-1, and ACAT-2. Compounds 2-5 showed varied degree of antioxidant activity on LDL with $IC_{50}$ values of 2.1, 11.8, 15.3, and $4.1{\mu}M$. Compounds 1, 2, and 3 showed inhibition activity on ACAT-1 with values $63.4{\pm}6.9%$ ($IC_{50}=66.8{\mu}M$), $53.7{\pm}0.9%$ ($IC_{50}=109.2{\mu}M$), and $78.7{\pm}0.2%$ ($IC_{50}=40.6{\mu}M$), respectively, at a concentration of 50 mg/mL, and on ACAT-2 with values $47.3{\pm}1.5%$ ($IC_{50}=149.7{\mu}M$), $39.2{\pm}0.2%$ ($IC_{50}=165.2{\mu}M$), and $52.1{\pm}1.0%$ ($IC_{50}=131.0{\mu}M$, respectively, at a concentration of 50 mg/mL.

Qualitative and quantitative analysis of the saponins in Panax notoginseng leaves using ultra-performance liquid chromatography coupled with time-of-flight tandem mass spectrometry and high performance liquid chromatography coupled with UV detector

  • Liu, Fang;Ma, Ni;He, Chengwei;Hu, Yuanjia;Li, Peng;Chen, Meiwan;Su, Huanxing;Wan, Jian-Bo
    • Journal of Ginseng Research
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    • v.42 no.2
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    • pp.149-157
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    • 2018
  • Background: Panax notoginseng leaves (PNL) exhibit extensive activities, but few analytical methods have been established to exclusively determine the dammarane triterpene saponins in PNL. Methods: Ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC/Q-TOF MS) and HPLC-UV methods were developed for the qualitative and quantitative analysis of ginsenosides in PNL, respectively. Results: Extraction conditions, including solvents and extraction methods, were optimized, which showed that ginsenosides Rc and Rb3, the main components of PNL, are transformed to notoginsenosides Fe and Fd, respectively, in the presence of water, by removing a glucose residue from position C-3 via possible enzymatic hydrolysis. A total of 57 saponins were identified in the methanolic extract of PNL by UPLC/Q-TOF MS. Among them, 19 components were unambiguously characterized by their reference substances. Additionally, seven saponins of PNL-ginsenosides Rb1, Rc, Rb2, and Rb3, and notoginsenosides Fc, Fe, and Fd-were quantified using the HPLC-UV method after extraction with methanol. The separation of analytes, particularly the separation of notoginsenoside Fc and ginsenoside Rc, was achieved on a Zorbax ODS C8 column at a temperature of $35^{\circ}C$. This developed HPLC-UV method provides an adequate linearity ($r^2$ > 0.999), repeatability (relative standard deviation, RSD < 2.98%), and inter- and intraday variations (RSD < 4.40%) with recovery (98.7-106.1%) of seven saponins concerned. This validated method was also conducted to determine seven components in 10 batches of PNL. Conclusion: These findings are beneficial to the quality control of PNL and its relevant products.

High Throughput-compatible Screening of Anti-oxidative Substances by Insect Extract Library (약용곤충추출물 라이브러리를 이용한 항산화 활성의 초고속 검색)

  • Park, Ja-Young;Heo, Jin-Chul;An, Sang-Mi;Yun, Eun-Young;Han, Sang-Mi;Hwang, Jae-Sam;Kang, Seok-Woo;Yun, Chi-Young;Lee, Sang-Han
    • Food Science and Preservation
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    • v.12 no.5
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    • pp.482-488
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    • 2005
  • Oxidant stress is well-known for a pivotal parameter related to neuro-inflammatory diseases including Alzheimer's disease, Parkinson's disease, and ALS (Lou Gehrig's disease). In order to effectively screen for anti-inflammatory agents, we first established the infrastructure of high throughput screening for anti-oxidant agents from medicinal insect library extracted with water, methanol, ethanol, and dimethyl sulfoxide. By the screening system, we found that Tenodera angustipennis Saussure, Pyrocoela rupa Olivier and Papilio maackii Mntris had strong anti-oxidant activity. Moreover, Tenodera angustipennis Saussure and Tenodera aridifolia (Stoll) exhibited protection effects of cellular damage by treatment of an oxidant hydrogen peroxide. Together, the results suggest that some selected hits could be a potential agent against neuro-inflammation, although the in vivo studies should be clearly tested.

Evaluation of antioxidant, α-glucosidase inhibition and acetylcholinesterase inhibition activities of Allium hookeri root grown in Korea and Myanmar (국내 및 미얀마에서 재배된 삼채뿌리의 항산화, α-Glucosidase 저해 및 Acetylcholinesterase 저해 활성)

  • Park, Joo Young;Yoon, Kyung Young
    • Food Science and Preservation
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    • v.23 no.2
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    • pp.239-245
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    • 2016
  • This study was conducted to compare the functionality (antioxidant, anti-diabetic, and anti-dementia activities) of the methanol extract of Allium hookeri root grown in Korea (KR) and Myanmar (MR). The total polyphenol and flavonoid contents of KR and MR were 5.27 and 4.80 mg GAE/g, and 0.35 and 0.24 mg QE/g, respectively. KR contained significantly higher levels of total polyphenols and total flavonoids than those of MR (p<0.05). The IC50 values of KR and MR were 6.53 and 5.31 mg/mL, respectively, for DPPH radical scavenging activity. However, KR had a significantly higher ABTS radical scavenging activity, $Fe^{2+}$ chelating ability, and reducing power compared with those of MR (p<0.05). In the evaluation of anti-diabetic activity, KR showed significantly higher ${\alpha}-glucosidase$ inhibition activity than acarbose and MR at whole concentrations (p<0.05). KR and MR had acetylcholinesterase inhibition activities that of 51.44% and 44.33%, respectively, at a 50 mg/mL concentration. These results suggested that roots of A. hookeri, especially KR, could be useful in improving diabetic and dementia disorders due to their high antioxidant, anti-diabetic, and anti-dementia activities.

Effect of Dangguibohyultang and its combinations on apoptosis in human colorectal adenocarcinoma HCT116 cells (당귀보혈탕(當歸補血湯)의 배합비율에 따른 대장암 세포주 HCT116의 세포사멸 효과)

  • Kim, Byung-Wan;Yun, Hyun-Joung;Jeon, Hyeon-Suk;Yun, Hyung-Joong;Kim, Chang-Hyun;Park, Sun-Dong
    • The Korea Journal of Herbology
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    • v.21 no.2
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    • pp.37-46
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    • 2006
  • Objectives : The purpose of this study was to investigate the effect of Dangguibohyultang (DB) and its combination (DB-I; Astragali membraneus BUNGE : Angelica gigas NAKAI=5:1, DB-II; Astragali membraneus BUNGE:Angelica gigas NAKAI=1:1, DB-III; Astragali membraneus BUNGE:Angelica gigas NAKAI=1:5,) on apoptosis in human colorectal adenocarcinoma HCT116 cells. Methods : To study the cytotoxic effect of methanol extract of DB-I, DB-II and DB-III on HCT116 cells, the cell viability was determined by XTT reduction method and ttypan blue exclusion assay. To confirm the induction of apoptosis, the cleavage of poly ADP-ribose polymerase (PARP), a substrate for caspase-3 and a typical sign of apoptosis, and the activation of procaspase-3, -8 and -9 were examined by western blot analysis. Furthermore, DB-induced apoptosis was confirmed by DNA fragmentation. The release of cytochrome C from mitochondria to cytosol, the level of Bcl-2 and Bax, and the expressions of Raf/MEK/ERK were examined by western blot analysis. Results : DB-I and DB-II reduced proliferation of HCT116 cells in a dose-dependent manner. DB-I and DB-II decreased procaspase-3, -8, -9 levels in a dose-dependent manner and induced the clevage of PARP. DB-I and DB-II also triggered the mitochondrial apoptotic signaling by increasing the release of cytochrome C from mitochondria to cytosol, decreasing of anti-apoptotic Bcl-2, and increasing of pro-apoptotic Bax. DB-I and DB-II decreased the activation of Ras/Raf/MEK/ERK cascade in a dose-dependent manner. Conclusion : These results suggest that DB-I and DB-II induce apoptosis via mitochondrial pathway in HCT116 cells. Furthermore, Raf/MEK/ERK cascade is involved in DB-induced apoptosis. These results suggest that DB is potentially useful as a chemotherapeutic agent in human liver cancer.

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Establishment of the Successive Rearing Method of Cabbage Butterfly, Pieris rapae L. in a Room Condition (배추흰나비의 실내 계대사육법 확립)

  • 설광열;김남정
    • Korean journal of applied entomology
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    • v.40 no.2
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    • pp.131-136
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    • 2001
  • Cabbage butterfly, Pieris rapae was reared in a room to establish a year-round rearing system. The eggs oviposited by the parent fed on host plant showed 89.2% of hatchability and hatched in 3.9 days after oviposition. The larval period was 18.1 days under high temperature, long day condition ($25^{\circ}C$, 16L : 8D), showing 97.8% pupal ratio and emergence rate. However, under low temperature, short day condition ($21^{\circ}C$, 10L : 14D) the larval period extended to 23.6 days and the pupal ratio was 70%. All of those pupae went into diapause. The oviposition preference experiment on different hosts (Chinese cabbage, cabbage, tulip and kale) showed that hot-water extract was preferred over methanol extracts or squeezed raw juices. The host preference showed that Chinese cabbage was less preferred than the other three. The artificial ovipositing kit was constructed for the oviposition in a room. The 48-hours old eggs could be stored for 7 days at$ 5^{\circ}C$ and showed 70% of hatchability. Non-diapausing pupae could be stored for 30 days at 5 to $15^{\circ}C$, showing 85% of emergence rate. However, the pupae stored at $5^{\circ}C$ showed longer storage period and higher emergence rate. The systematic successive rearing method of cabbage butterfly in a room was completed, based on the above experiments.

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Protective effects of Seoritae Chungkukjang added with green tea powder against 3-morpholinosydnonimine-induced oxidative stress (녹차 첨가 서리태 청국장의 3-morpholinosydnonimine에 의한 산화적 스트레스 개선 효과)

  • Cho, Eun-Ju;Park, Hyun-Young;Lee, Sang-Hyun;Kim, Hyun-Young
    • Korean Journal of Agricultural Science
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    • v.42 no.4
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    • pp.407-414
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    • 2015
  • To increase antioxidative activity of Chungkukjang, the protective effect of Seoritae Chungkukjang (SC) added with green tea powder against oxidative stress was evaluated under the cellular system using LLC-$PK_1$ cells. The treatment of 3-morpholinosydnonimine showed increase in lipid peroxidation, and decrease in endogenous anti-oxidant enzymes activity and cell viability. The methanol extract of SC inhibited lipid peroxidation by 70.9%, and significantly increased cell viability up to more than 33.2%. In addition, it enhanced superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities. Particularly, the addition of green tea in SC exerted protective effect against oxidative stress by ONOO- through elevation in activities of SOD and GSH-Px, and inhibition of lipid peroxidation. More addition of green tea showed stronger protective activity. These results suggest that the addition of green tea to SC leads to the increase in the antioxidative effect of Chungkukjang through elevation in antioxidative enzyme activities and protection from lipid peroxidation.

Anticancer effect of Rheum Rhizoma on human liver cancer HepG2 cells (간암 세포주 HepG2에 대한 대황 추출물의 항암효과)

  • Yun, Hyun-Joung;Hwang, Seong-Goo;Yun, Hyung-Joong;Kim, Chang-Hyun;Seo, Gyo-Soo;Park, Won-Hwan;Park, Sun-Dong
    • The Korea Journal of Herbology
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    • v.21 no.4
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    • pp.27-36
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    • 2006
  • Objectives : This study was performed for the investigation of anticancer effects of methanol extract of Rheum Rhizoma (MeOH-RR) on a human liver cancer cell line (HepG2). Methods : To study the cytotoxic effect of MeOH-RR on HepG2 cells, the cell viability was determined by XTT reduction method and trypan blue exclusion assay. The cleavage of poly ADP-ribose polymerase (PARP), a substrate for caspase-3 and a typical sign of apoptosis, and the activation of procaspase-3, -8 and -9 were examined by western blot analysis. Furthermore, MeOH-RR-induced apoptosis was confirmed by DNA fragmentation. The release of cytochrome c from mitochondria to cytosol, the level of Bcl-2 and Bax were examined by western blot analysis. Results : MeOH-RR reduced proliferation of HepG2 cells in a dose-dependent manner at 24 h and 48 h treatment. MeOH-RR induced the activation of caspase-3, -8, and -9 and the cleavage of poly ADP-ribose polymerase (PARP), a substrate for caspase-3. Furthermore, treatment with MeOH-RR resulted in internucleosomal DNA fragmentation, evidenced by the formation of a DNA ladder on agarose gel, a hallmark of cells undergoing apoptosis. MeOH-RR downregulated Bcl-2, upregulated Bax, and increased the release of cytochrome c from the mitochondria into cytosol in a dose-dependent manner. Moreover, MeOH-RP increased caspase-3 activity. Conclusion : There results suggest that MeOH-RR induce apoptosis via mitochondrial pathway and caspase-3-dependent pathway in HepG2 cells. There results suggest that MeOH-RR is potentially useful as a chemotherapeutic agent in human liver cancer.

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Antioxidant and Antimicrobial Effects of Solvent Fractions from Smilax china L. Leaves (청미래덩굴 잎(Smilax china L.) 용매 분획물의 항산화 및 항균 효과)

  • Kim, Joo-Young;Seoung, Gi-Un;Chung, Shin-Kyo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.10
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    • pp.1614-1618
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    • 2014
  • To utilize Smilax china L. leaves as a natural preservatives, solvent fractions from crude methanol extract were prepared and investigated their antioxidant and antimicrobial activities against Staphylococcus aureus. Antioxidant activities were examined by 1,1-diphenyl-1-picrylhydrazyl radical scavenging and ferric ion reducing antioxidant power assays. Ethyl acetate fraction from Smilax china L. leaves exhibited the strongest antioxidant and antimicrobial activities among the fractions (P<0.05), as well as the highest total phenolic and total flavonoid contents. Caffeic acid, ferulic acid, quercetin, and kaempferol contents in the ethyl acetate fraction from Smilax china L. leaves were determined by TLC and HPLC. In aqueous phase, as well as the n-butanol and n-hexane fractions, quercetin, ferulic acid, and kaempferol were detected in small amounts. Ferulic acid, which showed the highest content, is the major active compound from Smilax china L. leaves.

Imyosan induces caspases-mediated apoptosis in human colorectal cancer HCT116 cells (이묘산(二妙散)에 의한 대장암 세포주 HCT116의 Caspases 활성화를 매개로 한 세포사멸)

  • Kim, Sun-Mo;Yun, Hyun-Jeung;Lee, Hyun-Woo;Kim, Pan-Jun;Lee, Chang-Hyun;Park, Won-Hwan;Park, Sun-Dong
    • Herbal Formula Science
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    • v.14 no.2
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    • pp.21-32
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    • 2006
  • The purpose of this study was to investigate the effect of Imyosan on apoptosis in human colorectal cancer HCT116 cells. Phellodendron amurense Rupr. and Atratylodes lancea D.C. compose Imyosan. First of all, to study the cytotoxic effect of methanol extract of Imyosan (IMS-MeOH) on HCT116 cells, the cells were treated with various concentrations of IMS-MeOH and then cell viability was determined by XTT reduction method. IMS-MeOH reduced viability of HCT116 cells in a dose and time-dependent manner. To confirm the induction of apoptosis, the c1eavage of poly ADP-ribose polymerase (PARP), a substrate for caspase-3 and a typical sign of apoptosis, and the activation of caspase-3, procaspase-8 and procaspase-9 were examined by western blot analysis. IMS-MeOH decreased procaspase-3, procaspase-8 and procaspase-9 levels in a dose-dependent manner and induced the clevage of PARP. IMS-MeOH triggered the mitochondrial apoptotic signaling by increasing the release of cytochrome c from mitochondria to cytosol. Furthermore, IMS-MeOH also downregulated the anti-apoptotic Bcl-2 and upregulated the pro-apoptotic-Bax. Therefore, these results suggest that IMS-MeOH induced HCT1l6 cell death through the mitochondrial pathway. To explore whether the activities of caspases was required for induction of apoptosis by IMS-MeOH, caspase-3, -8, -9 activity measured by using substrates, respectively. IMS-MeOH increased caspase-3, -8, -9 activity. Co-treatment with inhibitors of caspase-3, -8, -9 and IMS-MeOH significantly blocked IMS-MeOH-triggered apoptosis in HCT1l6 cells. These results suggest that IMS-MeOH induces caspases-mediated apoptosis.

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