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Insilico profiling of microRNAs in Korean ginseng (Panax ginseng Meyer)
Mathiyalagan, Ramya,Subramaniyam, Sathiyamoorthy,Natarajan, Sathishkumar,Kim, Yeon Ju,Sun, Myung Suk,Kim, Se Young,Kim, Yu-Jin,Yang, Deok Chun The Korean Society of Ginseng 2013 Journal of Ginseng Research Vol.37 No.2
MicroRNAs (miRNAs) are a class of recently discovered non-coding small RNA molecules, on average approximately 21 nucleotides in length, which underlie numerous important biological roles in gene regulation in various organisms. The miRNA database (release 18) has 18,226 miRNAs, which have been deposited from different species. Although miRNAs have been identified and validated in many plant species, no studies have been reported on discovering miRNAs in Panax ginseng Meyer, which is a traditionally known medicinal plant in oriental medicine, also known as Korean ginseng. It has triterpene ginseng saponins called ginsenosides, which are responsible for its various pharmacological activities. Predicting conserved miRNAs by homology-based analysis with available expressed sequence tag (EST) sequences can be powerful, if the species lacks whole genome sequence information. In this study by using the EST based computational approach, 69 conserved miRNAs belonging to 44 miRNA families were identified in Korean ginseng. The digital gene expression patterns of predicted conserved miRNAs were analyzed by deep sequencing using small RNA sequences of flower buds, leaves, and lateral roots. We have found that many of the identified miRNAs showed tissue specific expressions. Using the insilico method, 346 potential targets were identified for the predicted 69 conserved miRNAs by searching the ginseng EST database, and the predicted targets were mainly involved in secondary metabolic processes, responses to biotic and abiotic stress, and transcription regulator activities, as well as a variety of other metabolic processes.
Insilico profiling of microRNAs in Korean ginseng (Panax ginseng Meyer)
Ramya Mathiyalagan,Sathiyamoorthy Subramaniyam,Sathishkumar Natarajan,Yeon Ju Kim,Myung Suk Sun,Se Young Kim,Yu-Jin Kim,Deok Chun Yang 고려인삼학회 2013 Journal of Ginseng Research Vol.37 No.2
MicroRNAs (miRNAs) are a class of recently discovered non-coding small RNA molecules, on average approximately 21 nucleotides in length, which underlie numerous important biological roles in gene regulation in various organisms. The miRNA database (release 18) has 18,226 miRNAs, which have been deposited from different species. Although miRNAs have been identified and validated in many plant species, no studies have been reported on discovering miRNAs in Panax ginseng Meyer, which is a traditionally known medicinal plant in oriental medicine, also known as Korean ginseng. It has triterpene ginseng saponins called ginsenosides, which are responsible for its various pharmacological activities. Predicting conserved miRNAs by homologybased analysis with available expressed sequence tag (EST) sequences can be powerful, if the species lacks whole genome sequence information. In this study by using the EST based computational approach, 69 conserved miRNAs belonging to 44 miRNA families were identified in Korean ginseng. The digital gene expression patterns of predicted conserved miRNAs were analyzed by deep sequencing using small RNA sequences of flower buds, leaves, and lateral roots. We have found that many of the identified miRNAs showed tissue specific expressions. Using the insilico method, 346 potential targets were identified for the predicted 69 conserved miRNAs by searching the ginseng EST database, and the predicted targets were mainly involved in secondary metabolic processes, responses to biotic and abiotic stress, and transcription regulator activities, as well as a variety of other metabolic processes.
Mathiyalagan, Ramya,Subramaniyam, Sathiyamoorthy,Kim, Yeon Ju,Natarajan, Sathishkumar,Min, Jin Woo,Kim, Se Young,Yang, Deok Chun Japan Society for Bioscience, Biotechnology, and A 2014 Bioscience, Biotechnology, and Biochemistry Vol.78 No.3
<P>The ginsenosides in Panax ginseng have vast structural and pharmacological efficacies. We covalently conjugated polyethylene glycol on the surface of CK (PEG-CK) through an acid-labile ester-linkage that showed increased solubility of CK. HPLC analysis showed that the release of CK was enhanced at acidic pH 5, whereas it was dramatically decreased at physiological pH 7.4. This might enhance the efficacy of CK.</P>
Enhanced Efficacies of Ginsenoside Compound K conjugated with Polymers in cancer cells
Ramya Mathiyalagan,Sathiyamoorthy Subramaniyam,Prabhu Muthusamy,Deok Chun Yang 한의병리학회 2015 대한동의병리학회 학술대회논문집 Vol.2015 No.10
Cancer is a leading cause of death worldwide. Although numerous anticancer drugs are available, their lack of tumor-targeting and low water-solubility are major drawbacks to use in cancer treatment. Conjugation of anticancer drugs with water-soluble polymers has been exhibited increased water-solubility of hydrophobic drugs, reduced normal cell cytotoxicity, increased accumulation in tumor tissues by passive targeting, and enhanced permeation and retention (EPR) effect. This study, we used ginsenoside compound K (CK) which is triterpenoid found in oriental medical plant- Panax ginseng Meyer. CK is one of the major metabolite after oral administration of crude extracts, reaches systemic circulation and reported for various pharmacological activities. However, solubility and targeted delivery is the major drawback to use of CK in clinical trials. We attempted to developed polymer CK conjugates to enhance its solubility and targeted delivery. To this, hydrophobic CK was covalently conjugated to the hydrophilic glycol chitosan (GC) and polyethylene glycol (PEG) through an acid-labile ester linkage. The resulting GC-CK and PEG-CK conjugates were characterized by H NMR and FTIR. It formed self-assembled spherical shape nanoparticles in water. The conjugates were greatly increased solubility of CK. From in vitro release experiment, release of CK was enhanced under pH 5, whereas it was dramatically decreased under physiological conditions (pH 7.4) is similar to what would be expected in extracellular solid tumor tissues and intracellular endosomes and lysosomes. In vitro cytotoxicity assays revealed that conjugates exhibited higher cytotoxicity in HT29, HepG2, and HT22 cell lines and maintained good cell viability in RAW264.7 cells. These results suggest that these conjugates may be potentially useful as a tumor-specific delivery vehicle which may enhance the overall efficacy of CK.
Abbai, Ragavendran,Mathiyalagan, Ramya,Markus, Josua,Kim, Yeon-Ju,Wang, Chao,Singh, Priyanka,Ahn, Sungeun,Farh, Mohamed El-Agamy,Yang, Deok Chun DOVE MEDICAL PRESS 2016 INTERNATIONAL JOURNAL OF NANOMEDICINE Vol.11 No.-
<P>Pharmacologically active stem of the oriental herbal adaptogen, Siberian ginseng, was employed for the ecofriendly synthesis of Siberian ginseng silver nanoparticles (Sg-AgNPs) and Siberian ginseng gold nanoparticles (Sg-AuNPs). First, for metabolic characterization of the sample, liquid chromatography-tandem mass spectrometry analysis (indicated the presence of eleutherosides A and E), total phenol content, and total reducing sugar were analyzed. Second, the water extract of the sample mediated the biological synthesis of both Sg-AgNPs and Sg-AuNPs that were crystalline face-centered cubical structures with a Z-average hydrodynamic diameter of 126 and 189 nm, respectively. Moreover, Fourier transform infrared analysis indicated that proteins and aromatic hydrocarbons play a key role in the formation and stabilization of Sg-AgNPs, whereas phenolic compounds accounted for the synthesis and stability of Sg-AuNPs. 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide (MTT) assay determined that Sg-AgNPs conferred strong cytotoxicity against MCF7 (human breast cancer cell line) and was only slightly toxic to HaCaT (human keratinocyte cell line) at 10 µg⋅mL<SUP>−1</SUP>. However, Sg-AuNPs did not display cytotoxic effects against both of the cell lines. The disc diffusion assay indicated a dose-dependent increase in the zone of inhibition of <I>Staphylococcus aureus</I> (ATCC 6538), <I>Bacillus anthracis</I> (NCTC 10340), <I>Vibrio parahaemolyticus</I> (ATCC 33844), and <I>Escherichia coli</I> (BL21) treated with Sg-AgNPs, whereas Sg-AuNPs did not show inhibitory activity. In addition, the 2,2-diphenyl-1-picrylhydrazyl assay demonstrated that both Sg-AgNPs and Sg-AuNPs possess strong antioxidant activity. To the best of our knowledge, this is the first report unraveling the potential of <I>Eleutherococcus senticosus</I> for silver and gold nanoparticle synthesis along with its biological applications, which in turn would promote widespread usage of the endemic Siberian ginseng.</P>
Wang, Chao,Mathiyalagan, Ramya,Kim, Yeon Ju,Castro-Aceituno, Veronica,Singh, Priyanka,Ahn, Sungeun,Wang, Dandan,Yang, Deok Chun Dove Medical Press 2016 INTERNATIONAL JOURNAL OF NANOMEDICINE Vol.11 No.-
<P><I>Dendropanax morbifera</I> Léveille is an oriental medicinal plant that is traditionally used in folk medicine and grows in a specific region of South Korea. We aimed to enhance the utilization of <I>D. morbifera</I> medicinal plants for synthesis of silver nanoparticles (AgNPs) and gold nanoparticles (AuNPs). <I>D. morbifera</I> leaf extract acted as both a reducing and a stabilizing agent that rapidly synthesized <I>Dendropanax</I> AgNPs (D-AgNPs) and <I>Dendropanax</I> AuNPs (D-AuNPs). The D-AgNPs and D-AuNPs were characterized by ultraviolet-visible spectroscopy, energy dispersive X-ray analysis, elemental mapping, field emission transmission electron microscopy, X-ray diffraction, and dynamic light scattering. The characterizations revealed that the D-AgNPs and D-AuNPs were in polygon and hexagon shapes with average sizes of 100–150 nm and 10–20 nm, respectively. The important outcomes were the synthesis of AgNPs and AuNPs within 1 hour and 3 minutes, respectively, avoiding the subsequent processing for removal of any toxic components or for stabilizing the nanoparticles. Additionally, D-AgNPs and D-AuNPs were examined for cytotoxicity in a human keratinocyte cell line and in A549 human lung cancer cell line. The results indicated that D-AgNPs exhibited less cytotoxicity in the human keratinocyte cell line at 100 µg/mL after 48 hours. On the other hand, D-AgNPs showed potent cytotoxicity in the lung cancer cells at the same concentration after 48 hours, whereas D-AuNPs did not exhibit cytotoxicity in both cell lines at the same concentration. However, both D-AgNPs and D-AuNPs at 50 µg/mL enhanced the cytotoxicity of ginsenoside compound K at 25 µM after 48 hours of treatment compared with CK alone. We believe that this rapid green synthesis of D-AgNPs and D-AuNPs is a valuable addition to the applications of <I>D. morbifera</I> medicinal plant. D-AuNPs can be used as carriers for drug delivery and in cancer therapy due to their lack of normal cell cytotoxicity.</P>
Subramaniyam, Sathiyamoorthy,Mathiyalagan, Ramya,Jun Gyo, In,Bum-Soo, Lee,Sungyoung, Lee,Deok Chun, Yang Springer 2011 Plant cell reports Vol.30 No.11
<P>Gynosaponins (Gypenosides) are major phyto-chemicals in Gynostemma pentaphyllum (Thunb.), with similarities to the ginsenosides present in Panax ginseng. Gynosaponins are classified as terpenoid compounds. In G. pentaphyllum, 25% of the total gynosaponins are similar to ginsenosides. In this study, we analyzed the transcriptional levels of the G. pentaphyllum genome to identify secondary metabolite genes. The complete transcriptomes for the roots and leaves were obtained using a GS-FLX pyro-sequencer. In total, we obtained 265,340 and all reads were well annotated according to biological databases. Using insilico analysis, 84% of sequence were well annotated and we obtained most of the secondary metabolite genes that represent mono-, di-, tri- and sesquiterpenoids. From our EST, most of the terpenoid genes were noted, among those few similar genes were studied in P. ginseng and these transcripts will help to characterize more triterpenoid genes in G. pentaphyllum. Also help to compare P. ginseng and G. pentaphyllum at transcriptome level.</P>