• Title/Summary/Keyword: methylene chloride extract

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Radical Scavenging Potential of Hydrophilic Phlorotannins of Hizikia fusiformis

  • Siriwardhana, Nalin;Lee, Ki-Wan;Jeon, You-Jin
    • ALGAE
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    • v.20 no.1
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    • pp.69-75
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    • 2005
  • Hizikia fusiformis is well known edible brown seaweed both in Korea and Japan. It has been intensively studied due to its pronounced health benefits. In this study, the radical scavenging (antioxidative) activities of its hydrophilic phlorotannin constituents were studied. An aqueous extract/original extract (OE) of H. fusiformis was initially prepared with heat, enzymes and pH control treatments. Then the original extract was further fractionated (with methylene chloride and methanol) and crude hydrophilic phlorotannin extract (CHPE) was prepared. The radical scavenging activities of both OE and CHPE were determined by using electron spin resonance (ESR) spectrophotometric assays such as 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl and alkyl radical assays. The CHPE reported significantly (p < 0.05) higher total phenolic (phlorotannins) content (1.23 mg ${\cdot}ml^{-1}$) than that of the OE (0.21 mg ${\cdot}ml^{-1}$). Both OE and CHPE have reported good radical scavenging activities and those activities were dose-dependent. The CHPE have demonstrated significantly higher radical scavenging activities than that of the OE. In comparison, the DPPH radical (6 $\times$ 10$^{-5}$ M) scavenging activities of all the CHPE concentrations (0.25, 0.5 and 1 mg ${\cdot}ml^{-1}$) tested were significantly higher (37.3, 78.2 and 91.6%, respectively) than that of the OE counterparts (11.4, 34.6 and 61.7%, respectively). Alkyl radical scavenging percentages of CHPE at 0.0625, 0.125, 0.25 and 0.5 mg${\cdot}ml^{-1}$ were significantly higher (34.3, 69.2, 80.4 and 88.7%, respectively) than that of the OE (16.6, 41.4, 62.3 and 77.4%, respectively). The percentages of hydroxyl radical scavenging activities of CHPE at the concentrations of 0.25, 0.5 and 1 mg ${\cdot}ml^{-1}$ were 32.5, 59.4 and 84.2 % respectively. Moreover, the hydroxyl radical scavenging activity of OE was quite lower than that of the CHPE. Therefore, these results suggest that the hydrophilic phlorotannins of H. fusiformis are potential radical scavengers thus, a great source of antioxidative nutraceuticles.

Crude Extract and Solvent-Partitioned Fractions of the Halophyte Atriplex gmelinii Inhibit Adipogenesis in 3T3-L1 Preadipocytes (3T3-L1 지방전구세포에서 염생식물 Atriplex gmelinii의 조추출물과 용매 분획물의 지방세포분화 억제)

  • Jung Im Lee;Jung Hwan Oh;Chang-Suk Kong;Youngwan Seo
    • Journal of Korean Medicine for Obesity Research
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    • v.23 no.2
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    • pp.69-77
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    • 2023
  • Objectives: Atriplex gmelinii C. A. Meyer is a halophyte belonging to the Chenopodiaceae family, and its young leaves and stems are used as fodder for livestock. The aim of the present study was to investigate the effects of A. gmelinii extract and its solvent fractions on lipid accumulation during adipogenesis of 3T3-L1 preadipocytes. Methods: The samples of A. gmelinii were separately extracted using methylene chloride and methanol. Subsequently, they were combined to formulate the initial extract, which was then partitioned based on polarity to prepare solvent fractions. Oil Red O staining was employed to measure lipid accumulation during the differentiation of 3T3-L1 preadipocytes. To verify cytotoxicity in 3T3-L1 cells, MTT assays were conducted. The expression levels of transcription factors in 3T3-L1 preadipocytes were measured through Western blotting analysis. Results: At 50 ㎍/mL, treatment of A. gmelinii extract and its solvent fractions during the differentiation of 3T3-L1 preadipocytes significantly diminished lipid accumulation with no noteworthy cytotoxicity on cell viability. Additionally, when investigating the biochemical pathways that underlie the prevention of lipid accumulation using solvent fractions, it was found that the n-BuOH and n-hexane fractions significantly decreased the expression of key transcription factors involved in the generation of fat, such as peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding protein α (C/EBPα), and sterol regulatory element-binding protein-1c (SREBP1c). Conclusions: These findings indicate that A. gmelinii can effectively reduce the accumulation of fat in 3T3-L1 adipocytes, making it a potentially valuable material for mitigating and preventing obesity.

Experimental Study of Citri Reticulatae Viride Pericarpium extract on the Cerebral Hemodynamics in Rats (청피가 뇌혈류역학에 미치는 실험적 연구)

  • Lee Geum Soo;Jeong Hyun Woo;Lee Won Suk
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.1
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    • pp.104-110
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    • 2002
  • Citri Reticulatae Viride Pericarpium extract(CRVP) have been used in oriental medicine for many centuries as a therapeutic agent for Soothing the liver and regulating the circulation of qi(疏肝理氣), and promoting digestion and removing stagnated food(消積化滯). The effects of CRVP on the vascular system is not known. The purpose of this Study was to investigate the effects of CRVP on the pial arterial diameter and regional cerebral blood flow(rCBF) in normal rats and ischemic cerebrovascular pathologic model rats. The changes in rCBF was determinated by Laser-Doppler Flowmetry(LDF), and the changes in pial arterial diameter were determinated by video microscopy methods and video analyzer. The results were as follows ; 1. Pial arterial diameter was significantly increased by CRVP in a dose-dependent manner. 2. Pretreatment with L-NNA significantly inhibited CRVP induced increased rCBF and pial arterial diameter. 3. Both the methylene chloride fraction and the hexane fraction of CRVP dose-dependently improved the altered cerebral hemodynamics of cerebral ischemic animal by increasing rCBF. 4. Pretreatment with L-NNA and indomethacin significantly inhibited CRVP(MC) induced increased rCBF. 5. Pretreatment with L-NNA and indomethacin significantly inhibited CRVP(hexane) induced increased rCBF. 6. Pretreatment with CRVP maredly stabilized the changes rCBF and pial arterial diameter during the period of cerebral reperusion. In conclusion, CRVP causes a diverse response of rCBF and pial arterial diameter, and CRVP dose-dependently improved the altered cerebral hemodynamics of cerebral ischemic animal by increasing rCBF and pial arterial diameter. These results suggest that the improvement of cerebral hemodynamics is also mediated by nitric oxide synthase and cyclooxygenase.

In Vitro Radical Scavenging Effect and Neuroprotective Activity from Oxidative Stress of Petasites japonicus (머위 분획물의 In Vitro 라디칼 소거능 및 신경세포의 산화적 스트레스 보호 효과)

  • Wang, Qian;Lee, Ah Young;Choi, Ji Myung;Lee, Dong Gu;Kim, Hyun Young;Lee, Sanghyun;Cho, Eun Ju
    • Korean Journal of Pharmacognosy
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    • v.45 no.2
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    • pp.147-153
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    • 2014
  • This study was focused on the evaluation of radical scavenging effect and the protective activity against oxidative stress of the extract and fractions from Petasites japonicus. P. japonicus was extracted with methanol and then fractionated into 4 fractions [n-butanol, ethyl acetate (EtOAc), methylene chloride, and n-hexane]. The extract and fractions showed strong 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity. Among all the fractions, particularly, the EtOAc fraction showed the strongest effect with the $IC_{50}$ value of $0.02{\mu}g/ml$. In addition, the fractions also showed strong hydroxyl radical scavenging activity and nitric oxide scavenging activity as well. Furthermore, cell viability generated by the P. japonicus extract and 4 fractions were examined under C6 glial cellular model. The C6 glial cells showed high generation of reactive oxygen species (ROS) and decrease in cell viability by the treatment generator of hydrogen peroxide. However, the production of ROS formation was decreased by the treatment of the fractions of P. japonicus and also founded that the EtOAc fraction led to significant increase in the cell viability at concentration $100{\mu}g/ml$. Results from this work indicated that P. japonicus showed protective effects against oxidative stress and its EtOAc fraction may be served as a useful natural antioxidant.

Inhibitory Effect of Lithospermum erythrorhizon Extracts on Melanin Biosynthesis (지치(Lithospermum erythrorhizon) 추출물의 멜라닌 생합성 억제효과)

  • Lee, Hwanghee Blaise;Bai, Suk;Chin, Jong-Eon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.9
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    • pp.1325-1329
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    • 2005
  • To estimate the inhibitory effect of Lithospemum erythrorhizon root extract on melanin biosynthesis, we tested its inhibitory effects on tyrosinase promoter in B16 mouse melanoma cells. Lithospermum erthrorhizon root extract had inhibitory effect above $33\%$ on tyrosinase promoter at $10{\mu}g/mL$ and exhibited no cytotoxicity under $100{\mu}g/mL$. Also, melanin biosynthesis decreased approximately $11\%$ and $24\%$ at $10{\mu}g/mL$ and $100{\mu}g/mL$, respectively. Therefore, Lithospermum erythrorhizon root extract would be considered very effective regulator of tyrosinase promoter and melanin biosynthesis.

Anti-proliferative Components in the Roots Extract from Pueraria thunbergiana (갈근으로부터 분리한 암세포 증식저해물질)

  • Baek, Mok-Ryeon;Choi, Yeon-Hee;Yoo, Dae-Seok;Kim, Mi-Ri;Choi, Sang-Un;Hong, Kyung-Sik;Kim, Young-Sup;Kim, Young-Kyoon;Lee, Kang-Ro;Ryu, Shi-Yong
    • Korean Journal of Pharmacognosy
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    • v.40 no.1
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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.

Anti-allergic and Anti-inflammatory Effect of Leonurus sibiricus Seed Ethyl Acetate Fractions (충울자 EtOAc 분획의 항알러지 및 항염증 효과)

  • Jung, Yujung;Jeon, Youngsic;Kim, Hyung Ja;Kang, Ki Sung;Kim, Yong Kee;Kim, Su-Nam
    • YAKHAK HOEJI
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    • v.58 no.5
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    • pp.294-299
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    • 2014
  • In this study, we investigate anti-allergic and anti-inflammatory effects of Leonurus sibiricus seed (LSS) extract in basophilic leukemia RBL-2H3 cells. To identify anti-allergic actions of LSS, the degranulation was evaluated in IgE and DNP-BSA stimulated RBL-2H3 cells. At the concentration of $100{\mu}g/ml$ of methanol (MeOH) extract and Methylene chloride (MC) and Ethyl acetate (EtOAc) fractions, the degranulation was significantly inhibited 16.7%, 16.7% and 27.9% respectively. And then, to assess anti-inflammatory effects of LSS, IL-4 and IL-13 mRNA level were detected in PMA/ionomycin (PI)-induced RBL-2H3 cells and cell proliferation and IL-4 mRNA level in isolated splenocytes from Balb/c mice. LSS MeOH extract and MC and EtOAc fractions significantly decreased the level of IL-4 and IL-13 mRNA in PI-induced RBL-2H3 cells and showed inhibitory effects on cell proliferation and expression of IL-4 mRNA level in mouse splenocytes. Taken together, these results suggest that LSS has potential anti-allergic and anti-inflammatory effects and EtOAc fraction is the most effective in regulating immune responses.

Inhibitory Effects of Saururus Chinensis Extracts on Osteoclast Differentiation

  • Shim, Ki-Shuk;Kim, Soon-Nam;Kim, Myung-Hee;Kim, Young-Sup;Ryu, Shi-Yong;Min, Yong-Ki;Kim, Seong-Hwan
    • Natural Product Sciences
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    • v.14 no.2
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    • pp.113-117
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    • 2008
  • Saururus chinensis is a commonly used folk herb for the treatment of edema and liver diseases in Korea. To study the biological activity of Saururus chinensis in bone metabolism, we evaluated the effect of its extracts on osteoclast differentiation in vitro using primary mouse bone marrow-derived macrophages. Methanol extract (ME) from dried roots of Saururus chinensis was partitioned into methylene chloride (MF), ethyl acetate (EF), n-butanol (BF) and water fractions (WF). Tartrate-resistance acid phosphatase (TRAP) activity assay and western blot analysis were performed to determine the effect on osteoclast differentiation and mitogen-activated protein (MAP) kinases activation. ME, MF and EF dramatically inhibited receptor activator of ${NF-kB}$ ligand (RANKL)-induced formation of multinucleated osteoclasts and activation of MAP kinases. This study firstly demonstrated that ME, MF and EF of Saururus chinensis have the potential to inhibit the osteoclast differentiation, which results from the inhibition of MAP kinases activations in part.

Antioxidant Activity of the Halophyte Ligustrum japonicum (염생식물 광나무(Ligustrum japonicum)의 항산화 활성)

  • Baek, Seung Oh;Kim, Hojun;Jeong, Heejeong;Ju, Eunsin;Kong, Chang-Suk;Seo, Youngwan
    • KSBB Journal
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    • v.30 no.6
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    • pp.275-282
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    • 2015
  • Dried samples of Ligustrum japonicum were extracted twice: with methylene chloride and with methanol (MeOH), respectively. The combined crude extracts were successively fractionated into n-hexane, 85% aqueous methanol (85% aq.MeOH), n-butanol (n-BuOH), and water fractions by liquid-liquid partition. Antioxidant activities of crude extract and its solvent fractions were evaluated by measuring authentic $ONOO^-$, and $ONOO^-$ generated from 3-morpholinsydnonimine (SIN-1) as well as degree of occurrence of intracellular ROS in HT 1080 cells, and genomic DNA oxidation. The 85% aq.MeOH and n-BuOH fractions exhibited the good antioxidant activity. Further purification of the 85% aq.MeOH fracition led to the isolation of Oleanolic acid (1), Maslinic acid (2), and Ursolic acid (3). All compounds showed the significant antioxidant effects in all assay systems.

Scutellaria baicalensis Georgi Extracts inhibit RANKL-induced Osteoclast Differentiation

  • Shim, Ki-Shuk;Kim, Soon-Nam;Kim, Myung-Hee;Kim, Young-Sup;Ryu, Shi-Yong;Min, Yong-Ki;Kim, Seong-Hwan
    • Natural Product Sciences
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    • v.14 no.3
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    • pp.182-186
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    • 2008
  • Scutellaria baicalensis Georgi (SBG) is traditionally used medicinal herb that has anti-oxidant, anticancer and anti-inflammatory effects. In this study, we investigated whether the extracts of SBG have the inhibitory activity in the osteoclast differentiation by using mouse monocytes RAW264.7 cells and primary mouse bone marrow-derived macrophages (BMMs). Methanol extract (ME) from SBG was successively fractionated into methylene chloride (MF), ethylacetate (EF) and n-butanol fraction (BF). The activity assay for tartrateresistant acid phosphatase (TRAP) and Western blot analysis were employed to evaluate the osteoclasts differentiation and the activation of mitogen-activated protein (MAP) kinases, respectively. ME, MF, EF and BF significantly and dose-dependently inhibited osteoclast differentiation without the decrease of cell viability at the concentrations used in this study. In addition, ME significantly inhibited the activation of c-jun-N-terminal kinase (JNK). In conclusion, this study firstly demonstrated that ME of SBG has the potential to inhibit the osteoclast differentiation through the suppression of JNK activation partially.