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Analysis of the Marker Compounds in Sagunja-tang by LC-ESI-MS (LC-ESI-MS에 의한 사군자탕의 지표성분 분석)

  • Seo, Chang-Seob;Shin, Hyeun-Kyoo
    • Korean Journal of Pharmacognosy
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    • v.50 no.1
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    • pp.65-71
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    • 2019
  • One of the oriental medicine prescriptions, Sagunja-tang consists of four herbal medicines (Ginseng Radix, Poria Sclerotium, Atractylodis Rhizoma Alba, and Glycyrrhiziae Radix et Rhizoma) and has been used as a medicine to enhance tonify the function of spleen and stomach in Korea. In this study, we conducted simultaneous analysis of the 9 marker components, liquiritin apioside, liquiritin, ginsenoside Rg1, liquiritigenin, ginsenoside Rb1, glycyrrhizin, atractylenolide III, atractylenolide II, and atractylenolide I in Sagunja-tang using a liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS/MS). Marker compounds were separated on a Waters Acquity UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, 1.7 mm) and the column was maintained at $45^{\circ}C$. The mobile phase consists of 0.1% (v/v) aqueous formic acid and acetonitrile with gradient condition. The LC-MS analysis was performed using a Waters ACQUITY TQD LC-MS/MS system with multiple reaction monitoring (MRM) method in the positive and negative modes. The calibration curves of the nine marker components showed good linearity with coefficient of determination ${\geq}0.9984$ within tested range. The limits of detection and limits of quantification values were 0.27-2.42 ng/mL and 0.81-7.27 ng/mL, respectively. The concentrations of tested 9 analytes in the lyophilized Sagunja-tang sample using the established LC-ESI-MS/MS MRM method were detected up to 16.593 mg/g. These results can be useful as a basic data for the quality control of an oriental medicine prescriptions.

Suppressor of Variegation 3-9 Homolog 2, a Novel Binding Protein of Translationally Controlled Tumor Protein, Regulates Cancer Cell Proliferation

  • Kim, A-Reum;Sung, Jee Young;Rho, Seung Bae;Kim, Yong-Nyun;Yoon, Kyungsil
    • Biomolecules & Therapeutics
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    • v.27 no.2
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    • pp.231-239
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    • 2019
  • Suppressor of Variegation 3-9 Homolog 2 (SUV39H2) methylates the lysine 9 residue of histone H3 and induces heterochromatin formation, resulting in transcriptional repression or silencing of target genes. SUV39H1 and SUV39H2 have a role in embryonic development, and SUV39H1 was shown to suppress cell cycle progression associated with Rb. However, the function of human SUV39H2 has not been extensively studied. We observed that forced expression of SUV39H2 decreased cell proliferation by inducing $G_1$ cell cycle arrest. In addition, SUV39H2 was degraded through the ubiquitin-proteasomal pathway. Using yeast two-hybrid screening to address the degradation mechanism and function of SUV39H2, we identified translationally controlled tumor protein (TCTP) as an SUV39H2-interacting molecule. Mapping of the interacting regions indicated that the N-terminal 60 amino acids (aa) of full-length SUV39H2 and the C-terminus of TCTP (120-172 aa) were critical for binding. The interaction of SUV39H2 and TCTP was further confirmed by co-immunoprecipitation and immunofluorescence staining for colocalization. Moreover, depletion of TCTP by RNAi led to up-regulation of SUV39H2 protein, while TCTP overexpression reduced SUV39H2 protein level. The half-life of SUV39H2 protein was significantly extended upon TCTP depletion. These results clearly indicate that TCTP negatively regulates the expression of SUV39H2 post-translationally. Furthermore, SUV39H2 induced apoptotic cell death in TCTP-knockdown cells. Taken together, we identified SUV39H2, as a novel target protein of TCTP and demonstrated that SUV39H2 regulates cell proliferation of lung cancer cells.

Improving Strontium Isotope Ratio Analysis Using MC-ICP-MS (다검출기 유도결합 플라즈마 질량분석기를 이용한 스트론튬 동위원소비 분석법 개선)

  • Lee, Sin-Woo;Park, Jaeseon;Park, Hyun-Woo;Hwang, Jong Yeon;Kim, Kumhee;Chung, Hyun-Mi;Choi, Jong-Woo
    • Journal of Environmental Analysis, Health and Toxicology
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    • v.21 no.4
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    • pp.237-242
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    • 2018
  • Strontium (Sr) commonly exists in rock, groundwater, soil, plants, and animals. The Sr isotope ratio offers important information as a tracer on nature because the Sr isotopic composition is not fractionated by any biological process in these ecosystems. Hence, Sr isotope ratio has been used in several studies on tracing the Sr source for contaminated sites and human migration. In this study, we developed a separation method for Sr content, and then improved Sr isotope analysis using multi-collector inductively coupled plasma mass spectrometry (MC-ICP-MS). A powdered rock standard (NIST 2710a) was used to determine the removal of interference elements (Rb and Ca) and the recovery rate of Sr content. The results ranged from 98% to 106%. Additionally, three standard samples (NBS 987, IAPSO and NIST 1486) were analyzed to evaluate the precision and accuracy of the results. The measured $^{87}Sr/^{86}Sr$ ratio for all the samples were consistent with the reported values, within an error. These results indicate that our established Sr separation and Sr isotope measurement methods are reliable and can hence be useful in the fields of environmental and forensic sciences.

Ginsenosides attenuate bioenergetics and morphology of mitochondria in cultured PC12 cells under the insult of amyloid beta-peptide

  • Kwan, Kenneth Kin Leung;Yun, Huang;Dong, Tina Ting Xia;Tsim, Karl Wah Keung
    • Journal of Ginseng Research
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    • v.45 no.4
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    • pp.473-481
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    • 2021
  • Background: Mitochondrial dysfunction is one of the significant reasons for Alzheimer's disease (AD). Ginsenosides, natural molecules extracted from Panax ginseng, have been demonstrated to exert essential neuroprotective functions, which can ascribe to its anti-oxidative effect, enhancing central metabolism and improving mitochondrial function. However, a comprehensive analysis of cellular mitochondrial bioenergetics after ginsenoside treatment under Aβ-oxidative stress is missing. Methods: The antioxidant activities of ginsenoside Rb1, Rd, Re, Rg1 were compared by measuring the cell survival and reactive oxygen species (ROS) formation. Next, the protective effects of ginsenosides of mitochondrial bioenergetics were examined by measuring oxygen consumption rate (OCR) in PC12 cells under Aβ-oxidative stress with an extracellular flux analyzer. Meanwhile, mitochondrial membrane potential (MMP) and mitochondrial dynamics were evaluated by confocal laser scanning microscopy. Results: Ginsenoside Rg1 possessed the strongest anti-oxidative property, and which therefore provided the best protective function to PC12 cells under the Aβ oxidative stress by increasing ATP production to 3 folds, spare capacity to 2 folds, maximal respiration to 2 folds and non-mitochondrial respiration to 1.5 folds, as compared to Aβ cell model. Furthermore, ginsenoside Rg1 enhanced MMP and mitochondrial interconnectivity, and simultaneously reduced mitochondrial circularity. Conclusion: In the present study, these results demonstrated that ginsenoside Rg1 could be the best natural compound, as compared with other ginsenosides, by modulating the OCR of cultured PC12 cells during oxidative phosphorylation, in regulating MMP and in improving mitochondria dynamics under Aβ-induced oxidative stress.

Flow of MHD Powell-Eyring nanofluid: Heat absorption and Cattaneo-Christov heat flux model

  • Sharif, Humaira;Khadimallah, Mohamed A.;Naeem, Muhammad Nawaz;Hussain, Muzamal;Hussain, Sajjad;Tounsi, Abdelouahed
    • Advances in nano research
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    • v.10 no.3
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    • pp.221-234
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    • 2021
  • During the previous few years, phenomenon of bioconvection along with the use of nanoparticles showed large number of applications in technological and industrial field. This paper analyzed the bioconvection phenomenon in magnetohydrodynamic boundary layer flow of a Powell-Eyring nanoliquid past a stretchable cylinder with Cattaneo-Christov heat flux. In addition, the impacts of chemical reaction and heat generation/absorption parameter are considered. By the use of appropriate transformation, the governing PDEs (nonlinear) have been transformed and formulated into nonlinear ODEs. The resulting nonlinear ODEs subjected to relevant boundary conditions are solved analytically through homotopy analysis method which is programmed in Mathematica software. Graphical and numerical results versus physical quantities like velocity, temperature, concentration and motile microorganism are investigated under the impact of physical parameters. It is noted that velocity profile enhances as the curvature parameter A and Eyring-Powell fluid parameter M increases but a decline manner for large values of buoyancy ratio parameter Nr and bio-convection Rayleigh number Rb. In the presence of Prandtl number Pr, Eyring-Powell fluid parameter M and heat absorption parameter ��, temperature profile decreases. Nano particle concentration profile increases for increasing values of magnetic parameter Ha and thermophoresis parameter Nt. The motile density profile has revealed a decrement pattern for higher values of bio-convection Lewis number Lb and bio-convection peclet number Pe. This study may find uses in bio-nano coolant systems, advance nanomechanical bio-convection energy conversion equipment's, etc.

Correlation between the Content and Pharmacokinetics of Ginsenosides from Four Different Preparation of Panax Ginseng C.A. Meyer in Rats

  • Jeon, Ji-Hyeon;Lee, Jaehyeok;Lee, Chul Haeng;Choi, Min-Koo;Song, Im-Sook
    • Mass Spectrometry Letters
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    • v.12 no.1
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    • pp.16-20
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    • 2021
  • We aimed to compare the content of ginsenosides and the pharmacokinetics after the oral administration of four different ginseng products at a dose of 1 g/kg in rats. The four different ginseng products were fresh ginseng extract, red ginseng extract, white ginseng extract, and saponin enriched white ginseng extract prepared from the radix of Panax ginseng C.A. Meyer. The ginsenoside concentrations in the ginseng product and the rat plasma samples were determined using a liquid chromatography-tandem mass spectrometry (LC-MS/MS). Eight or nine ginsenosides of the 15 tested ginsenosides were detected; however, the content and total ginsenosides varied depending on the preparation method. Moreover, the content of triglycosylated ginsenosides was higher than that of diglycosylated ginsenosides, and deglycosylated ginsenosides were not present in any preparation. After the single oral administrations of four different ginseng products in rats, only four ginsenosides, such as 20(S)-ginsenosides Rb1 (GRb1), GRb2, GRc, and GRd, were detected in the rat plasma samples among the 15 ginsenosides tested. The plasma concentrations of GRb1, GRb2, GRc, and GRd were different depends on the preparation method but pharmacokinetic features of the four ginseng products were similar. In conclusion, a good correlation between the area under the concentration curve and the content of GRb1, GRb2, and GRc, but not GRd, in the ginseng products was identified and it might be the result of their higher content and intestinal biotransformation of the ginseng product.

Improvement of antithrombotic activity of red ginseng extract by nanoencapsulation using chitosan and antithrombotic cross-linkers: polyglutamic acid and fucoidan

  • Kim, Eun Suh;Lee, Ji-Soo;Lee, Hyeon Gyu
    • Journal of Ginseng Research
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    • v.45 no.2
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    • pp.236-245
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    • 2021
  • Background: Red ginseng (RG) extract, especially ginsenoside Rg1 and Rb1 fractions has been reported to have antithrombotic activities. However, gastric instability and low intestinal permeability are considered to be obstacles to its oral administration. We hypothesized that stability, permeability, and activities of RG might be improved by encapsulation within nanoparticles (NPs) prepared with antithrombotic coating materials. Methods: RG-loaded chitosan (CS) NPs (PF-NPs) were prepared by complex ionic gelation with the antithrombotic wall materials, polyglutamic acid (PGA), and fucoidan (Fu). The concentrations of PGA (mg/mL, X1) and Fu (mg/mL, X2) were optimized for the smallest particle size by response surface methodology. Antithrombotic activities of RG and PF-NPs were analyzed using ex vivo and in vivo antiplatelet activities, in vivo carrageenan-induced mouse tail, and arteriovenous shunt rat thrombosis models. Results: In accordance with a quadratic regression model, the smallest PF-NPs (286 ± 36.6 nm) were fabricated at 0.628 mg/mL PGA and 0.081 mg/mL Fu. The inhibitory activities of RG on ex vivo and in vivo platelet aggregation and thrombosis in in vivo arteriovenous shunt significantly (p < 0.05) increased to approximately 66.82%, 35.42%, and 38.95%, respectively, by encapsulation within PF-NPs. For an in vivo carrageenan-induced mouse tail thrombosis model, though RG had a weaker inhibitory effect, PF-NPs reduced thrombus significantly due to the presence of PGA and Fu. Conclusion: PF-NPs contributed to improve the activities of RG not only by nanoencapsulation but also by antithrombotic coating materials. Therefore, PG-NPs can be suggested as an efficient delivery system for oral administration of RG.

Ginsenosides repair UVB-induced skin barrier damage in BALB/c hairless mice and HaCaT keratinocytes

  • Li, Zhenzhuo;Jiang, Rui;Wang, Manying;Zhai, Lu;Liu, Jianzeng;Xu, Xiaohao;Sun, Liwei;Zhao, Daqing
    • Journal of Ginseng Research
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    • v.46 no.1
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    • pp.115-125
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    • 2022
  • Background: Ginsenosides (GS) have potential value as cosmetic additives for prevention of skin photoaging. However, their protective mechanisms against skin barrier damage and their active monomeric constituents are unknown. Methods: GS monomer types and their relative proportions were identified. A UVB-irradiated BALB/c hairless mouse model was used to assess protective effects of GS components on skin epidermal thickness and transepidermal water loss (TEWL). Skin barrier function, reflected by filaggrin (FLG), involucrin (IVL), claudin-1 (Cldn-1), and aquaporin 3 (AQP3) levels and MAPK phosphorylation patterns, were analyzed in UVB-irradiated hairless mice or HaCaT cells. Results: Total GS monomeric content detected by UPLC was 85.45% and was largely attributed to 17 main monomers that included Re (16.73%), Rd (13.36%), and Rg1 (13.38%). In hairless mice, GS ameliorated UVB-induced epidermal barrier dysfunction manifesting as increased epidermal thickness, increased TEWL, and decreased stratum corneum water content without weight change. Furthermore, GS treatment of UVB-irradiated mice restored protein expression levels and epidermal tissue distributions of FLG, IVL, Cldn-1, and AQP3, with consistent mRNA and protein expression results obtained in UVB-irradiated HaCaT cells (except for unchanging Cldn-1 expression). Mechanistically, GS inhibited JNK, p38, and ERK phosphorylation in UVB-irradiated HaCaT cells, with a mixture of Rg2, Rg3, Rk3, F2, Rd, and Rb3 providing the same protective MAPK pathway inhibition-associated upregulation of IVL and AQP3 expression as provided by intact GS treatment. Conclusion: GS protection against UVB-irradiated skin barrier damage depends on activities of six ginsenoside monomeric constituents that inhibit the MAPK signaling pathway.

Comparison of Phytochemical Constituents According to the Cultivation Method (Paddy Field Cultivation, Upland Field Cultivation) of Korean Fresh Ginseng (한국산 수삼의 재배환경(논, 밭)에 따른 식물화학성분 비교)

  • Her, Jae Young;Kim, Do Hyeong;Hwang, Yu Jin;Ko, Sung Kwon;Yang, Byung Wook
    • Journal of Korean Medicine for Obesity Research
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    • v.22 no.1
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    • pp.21-29
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    • 2022
  • Objectives: This study aimed to examine the changes in ingredients according to its cultivation method by examining the content of saponin and non-saponin components of ginseng. Methods: Ginseng saponin component analysis was compared and reviewed using the high-performance liquid chromatography method, and acidic polysaccharide component was measured using the carbazole sulfuric acid method. Results: The comparative analysis of ginseng saponin content of 4 and 6 years old fresh ginseng showed the following results. According to the cultivation method, upland field cultivation fresh ginseng showed higher average content of crude saponin than paddy field cultivation fresh ginseng. Whereas, paddy field cultivation fresh ginseng showed higher average content of total saponin than upland field cultivation fresh ginseng. Ginsenoside Rb1 showed higher content of paddy field cultivation fresh ginseng than upland field cultivation fresh ginseng in 6 years old ginseng. However, it showed higher content of upland field cultivation fresh ginseng than paddy field cultivation fresh ginseng in 4 years old ginseng. Additionally, ginsenoside Rg1 showed higher content of paddy field cultivation fresh ginseng than upland field cultivation fresh ginseng in 6 years old ginseng, whereas upland field cultivation fresh ginseng showed higher content of paddy field cultivation fresh ginseng in 4 years old ginseng. The effect on the content of ginseng saponins and acidic polysaccharides according to paddy field and upland field cultivation methods is considered to be small. Conclusions: The paddy field cultivation method, which is more efficient in production cost due to depletion of farmland and long-distance cultivation, is considered to be an economical cultivation method.

Regulation of appetite-related neuropeptides by Panax ginseng: A novel approach for obesity treatment

  • Phung, Hung Manh;Jang, Dongyeop;Trinh, Tuy An;Lee, Donghun;Nguyen, Quynh Nhu;Kim, Chang-Eop;Kang, Ki Sung
    • Journal of Ginseng Research
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    • v.46 no.4
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    • pp.609-619
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    • 2022
  • Obesity is a primary factor provoking various chronic disorders, including cardiovascular disease, diabetes, and cancer, and causes the death of 2.8 million individuals each year. Diet, physical activity, medications, and surgery are the main therapies for overweightness and obesity. During weight loss therapy, a decrease in energy stores activates appetite signaling pathways under the regulation of neuropeptides, including anorexigenic [corticotropin-releasing hormone, proopiomelanocortin (POMC), cholecystokinin (CCK), and cocaine- and amphetamine-regulated transcript] and orexigenic [agoutirelated protein (AgRP), neuropeptide Y (NPY), and melanin-concentrating hormone] neuropeptides, which increase food intake and lead to failure in attaining weight loss goals. Ginseng and ginsenosides reverse these signaling pathways by suppressing orexigenic neuropeptides (NPY and AgRP) and provoking anorexigenic neuropeptides (CCK and POMC), which prevent the increase in food intake. Moreover, the results of network pharmacology analysis have revealed that constituents of ginseng radix, including campesterol, beta-elemene, ginsenoside Rb1, biotin, and pantothenic acid, are highly correlated with neuropeptide genes that regulate energy balance and food intake, including ADIPOQ, NAMPT, UBL5, NUCB2, LEP, CCK, GAST, IGF1, RLN1, PENK, PDYN, and POMC. Based on previous studies and network pharmacology analysis data, ginseng and its compounds may be a potent source for obesity treatment by regulating neuropeptides associated with appetite.