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Subash Karki,박희준,Agung Nugroho,김언지,정현아,최재수 한국식품영양과학회 2015 Journal of medicinal food Vol.18 No.1
The aim of the present study was to evaluate the comparative anti-inflammatory activities of Ixeris dentata (ID), Ixeris dentata var. albiflora (IDA), and Ixeris sonchifolia (IS) and to identify the main compounds present in extracts. The anti-inflammatory activity was evaluated through lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 murine macrophages. Five main compounds consisting of chlorogenic acid, caffeic acid, luteolin 7-O-glucoside, luteolin 7-O-glucuronide, and luteolin were used for simultaneous high-performance liquid chromatography quantification. The total phenolic content present in ID (30 mg/g GAE), IDA (35.33 mg/g GAE), and IS (43.79 mg/g GAE) was correlated to the corresponding LPS-induced NO production inhibitory effect in RAW 264.7 cells as expressed with IC50 values 26.19, 21.43, and 7.59 μg/mL, respectively. Luteolin 7-O-glucoside was found as the major compound in ID (8.76 mg/g dry weight) and IDA (10.35 mg/g dry weight) and luteolin 7-O-glucuronide was the major compound in IS (34.66 mg/g dry weight). Luteolin 7-O-glucoside and luteolin 7-O-glucuronide inhibited LPS-induced NO production with IC50 values of 30 and 4.5 μM, respectively. Furthermore, luteolin, luteolin 7-O-glucoside, and luteolin 7-O-glucuronide suppressed the expression of iNOS and COX-2, and t-BHP-induced ROS generation in LPS-stimulated RAW 264.7 cells. These results clearly showed that the anti-inflammatory potential of ID, IDA, and IS extract are primarily due to their contents of luteolin 7-O-glucoside and luteolin 7-O-glucuronide, respectively.
BACE1 and cholinesterase inhibitory activities of Nelumbo nucifera embryos
정현아,Subash Karki,김지혜,최재수 대한약학회 2015 Archives of Pharmacal Research Vol.38 No.6
The aim of the present study was to evaluatethe comparative anti-Alzheimer’s disease (AD) activitiesof different parts of Nelumbo nucifera (leaves, de-embryoseeds, embryos, rhizomes, and stamens) in order to determinethe selectivity and efficient use of its individualcomponents. Anti-AD activities of different parts of N. nucifera were evaluated via inhibitory activities on acetylcholinesterase(AChE), butyrylcholinesterase (BChE),and b-site amyloid precursor protein-cleaving enzyme 1(BACE1) along with scavenging activity on peroxynitrite(ONOO-). Among the evaluated parts of N. nucifera, theembryo extract exhibited significant inhibitory potentialagainst BACE1 and BChE as well as scavenging activityagainst ONOO-. Thus, the embryo extract was selected fordetailed investigation on anti-AD activity using BACE1-and ChEs-inhibitory assays. Among the different solventsolublefractions, the dichloromethane (CH2Cl2), ethylacetate (EtOAc), and n-butanol (n-BuOH) fractionsshowed promising ChEs and BACE1 inhibitory activities. Repeated column chromatography of the CH2Cl2, EtOAcand n-BuOH fractions yielded compounds 1–5, which wereneferine (1), liensinine (2), vitexin (3), quercetin 3-O-glucoside(4) and northalifoline (5). Compound 2 exhibitedpotent inhibitory activities on BACE1, AChE, and BChEwith respective IC50 values of 6.37 ± 0.13, 0.34 ± 0.02,and 9.96 ± 0.47 lM. Likewise, compound 1 showedpotent inhibitory activities on BACE1, AChE, and BChEwith IC50 values of 28.51 ± 4.04, 14.19 ± 1.46, and37.18 ± 0.59 lM, respectively; the IC50 values of 3 were19.25 ± 3.03, 16.62 ± 1.43, and 11.53 ± 2.21 lM,respectively. In conclusion, we identified potent ChEs- andBACE1-inhibitory activities of N. nucifera as well as itsisolated constituents, which may be further explored todevelop therapeutic and preventive agents for AD andoxidative stress related diseases.
Jung, Hyun Ah,Karki, Subash,Ehom, Na-Yeon,Yoon, Mi-Hee,Kim, Eon Ji,Choi, Jae Sue The Korean Society of Food Science and Nutrition 2014 Preventive Nutrition and Food Science Vol.19 No.4
The aim of the present study was to evaluate the anti-diabetic, anti-inflammatory, antioxidant potential, and total phenolic content (TPC) of green and red kohlrabi cultivars. Anti-diabetic and anti-inflammatory activities were evaluated via protein tyrosine phosphatase (PTP1B) and rat lens aldose reductase inhibitory assays and cell-based lipopolysaccharide (LPS)-induced nitric oxide (NO) inhibitory assays in RAW 264.7 murine macrophages. In addition, scavenging assays using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical, and peroxynitrite ($ONOO^-$) were used to evaluate antioxidant potential and TPC was selected to assess phytochemical characteristics. Between the two kohlrabi cultivars, red kohlrabi (RK) had two times more TPC than green kohlrabi (GK) and showed significant antioxidant effects in DPPH, ABTS, and $ONOO^-$ scavenging assays. Likewise, methanol (MeOH) extracts of RK and GK inhibited LPS-induced NO production in a dose dependent manner that was further clarified by suppression of iNOS and COX-2 protein production. The MeOH extracts of RK and GK exhibited potent inhibitory activities against PTP1B with the corresponding $IC_{50}$ values of $207{\pm}3.48$ and $287{\pm}3.22{\mu}g/mL$, respectively. Interestingly, the RK MeOH extract exhibited significantly stronger anti-inflammatory, anti-diabetic, and antioxidant effects than that of GK MeOH extract. As a result, our study establishes that RK extract with a higher TPC might be useful as a potent anti-diabetic, antioxidant, and anti-inflammatory agent.
Acetyl- and butyryl-cholinesterase inhibitory activities of the edible brown alga Eisenia bicyclis
최재수,Shourav Haulader,Subash Karki,정희진,김형락,정현아 대한약학회 2015 Archives of Pharmacal Research Vol.38 No.8
As part of our ongoing isolation of cholinesterase (ChE) inhibitors from natural marine sources, the bioactivity of the ethanolic extracts from 12 Korean seaweeds were screened for their inhibitory activities against acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and total reactive oxygen species (ROS) generation. Eisenia bicyclis exhibited promising inhibitory properties against AChE, BChE and total ROS with inhibition percentages (%) of 68.01 ± 1.37, 95.72 ± 3.80, and 73.20 ± 1.82 at concentrations of 25 lg/mL, respectively. Among the different solvent–soluble fractions obtained from the ethanolic extract,theethylacetate(EtOAc)fractionwasfoundtocause the most potent scavenging, or inhibitory activities, against 2,2-diphenyl-1-picrylhydrazyl (DPPH), peroxynitrite (ONOO-) and total ROS with the respective IC50 values of 2.48 ± 0.01, 8.70 ± 0.06, and 0.81 ± 0.03 lg/mL. Likewise, the EtOAc fraction also exhibited potent inhibitory activities against AChE and BChE with IC50 values of 2.78 ± 0.07 and 3.48 ± 0.32 lg/mL, respectively. Silica gel column chromatography of the EtOAc fraction yielded a phlorotannin, 974-B, based on the comparison with reported 1H- and 13C-NMR spectroscopic data. 974-B showed strong scavenging/or inhibitory potential against DPPH, ONOO-, total ROS, AChE, and BChE with the respective IC50 values of0.86 ± 0.02,1.80 ± 0.01,6.45 ± 0.04,1.95 ± 0.01,and 3.26 ± 0.08 lM, respectively. These results indicate thatthe potential of E. bicyclis and its phlorotannin for use in the development of therapeutic or preventive agents of Alzheimer’s disease mainly through ChE inhibition and additional antioxidant capacities.
Hyun Ah Jung,Subash Karki,Na-Yeon Ehom,Mi-Hee Yoon,Eon Ji Kim,Jae Sue Choi 한국식품영양과학회 2014 Preventive Nutrition and Food Science Vol.19 No.4
The aim of the present study was to evaluate the anti-diabetic, anti-inflammatory, antioxidant potential, and total phenolic content (TPC) of green and red kohlrabi cultivars. Anti-diabetic and anti-inflammatory activities were evaluated via protein tyrosine phosphatase (PTP1B) and rat lens aldose reductase inhibitory assays and cell-based lipopolysaccharide (LPS)-induced nitric oxide (NO) inhibitory assays in RAW 264.7 murine macrophages. In addition, scavenging assays using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical, and peroxynitrite (ONOO?) were used to evaluate antioxidant potential and TPC was selected to assess phytochemical characteristics. Between the two kohlrabi cultivars, red kohlrabi (RK) had two times more TPC than green kohlrabi (GK) and showed significant antioxidant effects in DPPH, ABTS, and ONOO? scavenging assays. Likewise, methanol (MeOH) extracts of RK and GK inhibited LPS-induced NO production in a dose dependent manner that was further clarified by suppression of iNOS and COX-2 protein production. The MeOH extracts of RK and GK exhibited potent inhibitory activities against PTP1B with the corresponding IC50 values of 207±3.48 and 287±3.22 ㎍/mL, respectively. Interestingly, the RK MeOH extract exhibited significantly stronger anti-inflammatory, anti-diabetic, and antioxidant effects than that of GK MeOH extract. As a result, our study establishes that RK extract with a higher TPC might be useful as a potent anti-diabetic, antioxidant, and anti-inflammatory agent.
Agung Nugroho,임상철,Subash Karki,최재수,박희준 한국생약학회 2015 Natural Product Sciences Vol.21 No.3
Peroxynitrite (ONOO-)-scavenging activities of nine Compositae herbs consisting of three Ixeris, two Youngia, two Cirsium and one of each Lactuca and Taraxacum species were evaluated. The contents of their ONOO- scavengers in the extracts were also determined on a HPLC using seven standard compounds, chlorogenic acid (CGA), chicoric acid (CA), luteolin 7-glucoside (luteolin-7-glc), luteolin 7-glucuronide (luteolin-7-glcU), luteolin, linarin and pectolinarin. Five of those compounds exhibited potent ONOO--scavenging activities: IC50, CA (0.76 mM), CGA (1.34 mM), luteolin (0.81 mM), luteolin-7-glc (0.86 mM) and luteolin-7-glcU (3.13 mM). Both CA and luteolin-7-glc were highly contained in I. dentata (19.71 mg/g and 13.58 mg/g, respectively), I. dentata var. albiflora (17.58 mg/g and 23.83 mg/g, respectively) and I. sonchifolia (65.71 mg/g and 6.99 mg/g, respectively). Among the nine herbs, those three Ixeris species had very low IC50 values over the range of 0.48 - 1.74 mg/mL, suggesting that they could be potential therapeutic vegetables, particularly for preventing diabetic complications or obesity, which can be caused by an excess production of ONOO-.