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사상 체질 처방의 항동맥경화 효능 비교 연구 -열다한소탕, 양격산화탕, 청심연자탕, 태음조위탕-
김온순,김예지,신현규,Kim, Ohn-Soon,Kim, Ye-Ji,Shin, Hyeun-Kyoo 사상체질의학회 2012 사상체질의학회지 Vol.24 No.4
Objectives : The purpose of this study was to investigate the anti-atherosclerotic effects of 4 herbal formulas for Sasang constitutional medicine (Yeoldahanso-tang: YDHST, Yanggyeoksanhwa-tang: YGSHT, Cheongsimyeonja-tang: CSYJT and Taeeumjowi-tang: TEJWT). Methods : The antioxidant activities of herbal formulas were studied by measuring free radical scavenging activities on ABTS and DPPH. The inhibitory effects on LDL oxidation was evaluated by the formation of TBARS, REM and fragmentation of apolipoprotein B-100 (ApoB). Effects of herbal formulas on macrophage lipid accumulation were determined in native LDL and LPS co-incubated macrophages using Oil Red O staining. Results : The scavenging activities on ABTS and DPPH of herbal formulas were increased in dose-dependent manner (YDHST>YGSHT>CSYJT>TEJWT). Herbal formulas reduced the oxidation properties of LDL induced by $CuSO_4$. YDHST, YGSHT and CSYJT showed strong suppressive effect on LDL oxidation than TEJWT. In addition, YDHST, YGSHT and CSYJT significantly inhibited foam cell formation in LDL/LPS stimulated RAW 264.7 cells. Conclusions : These results demonstrate that YDHST, YGSHT and CSYJT have potentials on anti-atherosclerosis by antioxidative effect and suppressive effect on LDL oxidation.
김온순,서창섭,신현규,Kim, Ohn Soon,Seo, Chang-Seob,Shin, Hyeun-Kyoo 대한예방한의학회 2013 대한예방한의학회지 Vol.17 No.3
Objective : The purpose of this study was to establish the simultaneous analysis for six compounds in Gumiganghwal-tang (GMGHT, Jiuweiqianghuo-tang) and to investigate the anti-atherosclerotic effects of GMGHT in vitro. Methods : The column for separation of six compounds was used Luna $C_{18}$ column and maintained at $40^{\circ}C$. The mobile phase for gradient elution consisted of two solvent systems, 1.0% acetic acid in water and 1.0% acetic acid in acetonitrile. The analysis was carried out at a flow rate of 1.0 mL/min with pothodiode array (PDA) detection at 254, 280, and 320 nm. The injection volume was 10 ${\mu}L$. The antioxidant activities of GMGHT were evaluated by measuring free radical scavenging activities on 2,2'-Azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) and 1-1-diphenyl-2-picrylhydrazyl (DPPH). The inhibitory effects on low-density lipoprotein (LDL) oxidation were evaluated by the formation of thiobarbituric acid relative substances (TBARS), relative electrophoretic mobility (REM), and fragmentation of apolipoprotein B (ApoB)-100. Results : Calibration curves were acquired with $r^2{\geq}0.9998$. The contents of liquiritin, ferulic acid, baicalin, baicalein, glycyrrhizin, and wogonin in GMGHT were 1.784, 1.693, 37.899, 0.258, 1.869, and 0.034 mg/g, respectively. The GMGHT showed the radical scavenging activity in a dose-dependent manner. The concentration required for 50% reduction ($RC_{50}$) against ABTS and DPPH radicals were 72.51 ${\mu}g/mL$ and 128.49 ${\mu}g/mL$. Furthermore, GMGHT reduced the oxidation properties of LDL induced by $CuSO_4$. Conclusion : HPLC-PDA is considered as an available and convenient method for quality control and standardization of GMGH and GMGHT has potentials on anti-atherosclerosis by anti-oxidative effect and suppressive effect on LDL oxidation.
김온순,서창섭,김예지,신현규,Kim, Ohn Soon,Seo, Chang-Seob,Kim, Yeji,Shin, Hyeun-Kyoo 대한한의학방제학회 2012 大韓韓醫學方劑學會誌 Vol.20 No.2
Objectives : The purpose of this study was to investigate the differences of chemical composition or biological activities between decoction and commercial herbal medicines of Samchulkunbi-tang (Shenzhujianpi-tang, SKT). Methods : The extracts of SKT from decoction (SKT1) and two different commercial extractive granules (SKT2 and SKT3) were prepared. The index components of SKTs were analyzed with HPLC. The antioxidant activities of SKTs were studied by measuring free radical scavenging activities on ABTS and DPPH. The anti-inflammatory effects were determined by measuring NO, $PGE_2$ and IL-6 in LPS-stimulated RAW 264.7 cells. Results : The contents of 7 components were 1.40-6.08 mg/g in SKT1, not detected-4.75 mg/g in SKT2, 0.03-1.46 mg/g in SKT3. The scavenging activities on ABTS and DPPH of herbal formulas were increased in dose-dependent manner (SKT1>SKT2>SKT3). SKT1 significantly inhibited $PGE_2$ and IL-6 production and SKT3 slightly inhibited $PGE_2$ production in LPS-stimulated RAW264.7 cells. SKT2 showed no inhibitory effects on production of inflammatory mediators such as $PGE_2$ and IL-6. Conclusions : These results demonstrate that the decoction of SKT has more strong anti-oxidant and anti-inflammatory effects than that of commercial herbal medicines consistent with the contents of index components.
LC-MS/MS를 이용한 이중탕(理中湯)의 정량분석 및 항산화 활성
서창섭,김온순,김예지,신현규,Seo, Chang-Seob,Kim, Ohn Soon,Kim, Yeji,Shin, Hyeun-Kyoo 대한한의학방제학회 2013 大韓韓醫學方劑學會誌 Vol.21 No.1
Objectives : Leejung-tang (Lizhong-tang) has been used for treatment of gastrointestinal disorders in Korea. In this study, we performed quantification analysis of five marker components, liquiritin, ginsenoside Rb1, ginsenoside Rg1, glycyrrhizin, and 6-gingerol in Leejung-tang using a ultra performance liquid chromatography- electrospray ionization-mass spectrometer (UPLC-ESI-MS). In addition, we evaluated antioxidant activity of Leejung- tang. Methods : The column for separation of five constituents used a UPLC BEH C18 ($100{\times}2.1mm$, $1.7{\mu}m$) maintained at $45^{\circ}C$. The mobile phase consisted of two solvent systems, 0.1% (v/v) formic acid in H2O (A) and CH3CN (B) by gradient flow. The flow rate was 0.3 mL/min with detection at mass spectrometer. The antioxidative activities conduct an experiment on 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities of Leejung-tang. Results : Calibration curves of five marker compounds were acquired with r2 values > 0.99. The amount of the five compounds in Leejung-tang were 0.07 - 0.84 mg/g. The concentration required for 50% reduction (RC50) against ABTS radical was 119.02 ug/mL. In addition, the scavenging against DPPH radical of Leejung-tang was 11.4%, 14.5%, 19.8%, 29.6%, and 49.2% at 25 ug/mL, $50{\mu}g/mL$, $100{\mu}g/mL$, $200{\mu}g/mL$, and $400{\mu}g/mL$, respectively. Conclusions : The established LC-MS/MS method will be helpful to improve quality control of Leejung-tang. In addition, Leejung-tang is a potential antioxidant therapeutic agent.
서창섭,김온순,신현규,Seo, Chang-Seob,Kim, Ohn Soon,Shin, Hyeun-Kyoo 대한한의학방제학회 2013 大韓韓醫學方劑學會誌 Vol.21 No.2
Objectives : We carry out the simultaneous quantification for quality control of four components in Bangpungtongseong-san (BPTSS) sample. In addition, we assessed the antioxidant effects of BPTSS sample. Methods : The used column for separation and analysis of four compounds was Luna C18 column and column oven temperature was maintained at $40^{\circ}C$. The mobile phase for simultaneous determination consisted of two solvent systems, 1.0% acetic acid in water and 1.0% acetic acid in acetonitrile. High performance liquid chromatography-photodiode array (HPLC-PDA) method for analysis was performed at a flow rate of 1.0 mL/min with PDA detection at 254 and 280 nm. The injection volume was 10 ${\mu}L$. The antioxidant activities of BPTSS were evaluated by measuring free radical scavenging activities on 2,2'-Azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) and 1-1-diphenyl-2-picrylhydrazyl (DPPH). The inhibitory effects on low-density lipoprotein (LDL) oxidation were evaluated by the formation of thiobarbituric acid relative substances (TBARS) and relative electrophoretic mobility (REM). Results : Calibration curves were acquired with $r^2{\geq}0.9999$. The values of limit of detection (LOD) and quantification (LOQ) were 0.06-0.29 ${\mu}g/mL$ and 0.20-0.98 ${\mu}g/mL$, respectively. The amounts of geniposide, liquiritin, baicalin, and glycyrrhizin in BPTSS were 5.06, 7.33, 27.56, and 7.81 mg/g, respectively. The BPTSS showed the radical scavenging activity in a dose-dependent manner. The concentration required for 50% reduction (RC50) against ABTS and DPPH radicals were 72.51 ${\mu}g/mL$ and 128.49 ${\mu}g/mL$. Furthermore, GMGHT reduced the oxidation properties of LDL induced by CuSO4. Conclusions : The established HPLC-PDA method will be helpful to improve quality control of BPTSS. In addition, BPTSS has potentials as therapeutic agent on anti-atherosclerosis.