• Korean Journal of Pharmacognosy
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• v.43 no.1
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• pp.66-71
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• 2012

Cirsium japonicum var. ussuriense is often used in treatment of human disease such as hemorrhage, blood congestion and inflammation. This study was accomplished to evaluate the antioxidant properties of the leaf (CLE) and flower (CFE) extracts of C. japonicum var. ussuriense to protect normal human red blood cells (RBC) and plasma samples against oxidative damage in vitro. CLE and CFE were prepared by extracting with hot water. In red blood cells and plasma, oxidative hemolysis and lipid peroxidation induced by the aqueous peroxyl radical generator [2,2'-azobis (2-amidinopropane) dihydrochloride, AAPH] were significantly suppressed by CLE or CFE in a dose-dependent manner at the same time. CLE and CFE also prevented the depletion of cytosolic antioxidant glutathione (GSH) in RBC. These results suggest that the leaves and flowers of C. japonicum var. ussuriense may have the antioxidant properties.

• Nutritional Sciences
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• v.9 no.1
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• pp.48-54
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• 2006

Methods have been developed to evaluate the total antioxidant capacity of foods and plasma but limitations are associated with their ability to determine precisely the contribution of lipophilic antioxidants in a lipid milieu as well as interactions among them Thus, we modified the Oxygen Radical Absorbance Capacity (ORAC) assay to determine the peroxyradical scavenging ability of both hydrophilic and lipophilic compartments in plasma The hydrophilic ORAC assay was performed in a phosphate buffer system utilizing 2,2'-azobis (2-amidinopropane) dihydrochloride as a peroxyradical generator and fluorescein as the target The lipophilic ORAC assay was carried out in a dimethylsulfoxide :butyronitrile (DMSO/BN, 9:1 v/v) system using 2,2'-azobis (2,4-dimethyl valeronitrile) as a peroxyradical generator and BODIPY C11 581/591 as the target Analyses were conducted in bovine serum supplemented with water - and lipid - soluble antioxidants and in human plasma. Albumin (0.5$\sim$5 g/dL) and uric acid (0.1$\sim$0.5 $\mu$mol/L) increased hydrophilic ORAC values in a dose-dependent fashion ($R^{2}$=0.97 and 0.98, respectively) but had no impact on lipophilic ORAC values. $\alpha$-Tocopherol (15$\sim$200 $\mu$mol/L) increased lipophilic ORAC values in a dose-dependent fashion ($R^{2}$=0.94); neither $\alpha$-tocopherol nor $\beta$-carotene had an impact on hydrophilic ORAC values. However, addition of $\beta$-carotene at physiological concentration (0.23$\sim$1.86 $\mu$mol/L), either alone or in combination with other carotenoids, had no significant impact on lipophilic ORAC values. Thus, while assays of 'total antioxidant capacity' in biological matrices would be a useful research and clinical tool, existing methods are limited by the lack of complete responsiveness to the full range of dietary antioxidants.

• Journal of Marine Bioscience and Biotechnology
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• v.10 no.2
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• pp.62-72
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• 2018

An active ingredient with hyaluronidase (HAse) inhibitory effect is one of the anti-aging approaches in cosmeceuticals. Here, red sea cucumbers (RSCs), Stichopus japonicus, from Jeju Island were evaluated to examine their HAse inhibitory and antioxidant activity effect. In this study, RSCs were extracted by six enzymatic hydrolysis (Alcalase; Al, Trypsin; Try, Neutrase; Neu, Pepsin; Pep, Alpha-chymotrypsin; Chy and Protamex; Pro). Alcalase hydrolysate (AlH) showed the highest antioxidant capacities for both of oxygen radical absorbance capacity (ORAC) and trolox equivalent antioxidant capacity (TEAC) methods, compared to those of other hydrolysates, at $66.59{\pm}0.78{\mu}M\;TE/mg$ and $135.78{\pm}3.24{\mu}M\;TE/mg$, respectively. Furthermore, AlH performed the highest capacity of HAse inhibitory with $IC_{50}$ value of 3.21 mg/ml. Thus, RSCs hydrolyzed by Al were chosen to determine the cellular antioxidant activity and hyaluronic acid (HA) production effect on Human immortalized keratinocyte cell line (HaCaT). The results showed that AlH improved the cell viabilities and intracellular reactive oxygen species (ROS) induced by 2,2'-Azobis(2-amidinopropane) dihydrochloride (AAPH) were significantly decreased. In addition, AlH increased HA amount by regulating HYAL2 and HAS2 expressions in the HaCaT cells. Taken together, AlH of RSCs collected from Jeju Island showed HAse inhibitory and antioxidant activities against skin-aging which shows its potentials can be an optional natural bioactive ingredient for novel cosmeceuticals.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.52 no.1
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• pp.35-42
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• 2019

We investigated the protective effects of antioxidant fractions from a 70% ethanolic extract of Hizikia fusiformis in human dermal fibroblasts (HDFs). Powdered H. fusiformis was extracted with 70% ethanol and then partitioned into three fractions according to polarity using n-hexane (HFH), chloroform (HFC), and ethyl acetate (HFEA). Antioxidant activity was observed in HFEA at 0.66 mg/mL based on the half maximal inhibitory concentration ($IC_{50}$) of 1,1-diphenyl-2-picrylhydrazyl (DPPH), and at 0.24 mg/mL based on alkyl radical scavenging. The protective effects of the HFEA antioxidant fraction against 2,2-azobis-(2-amidinopropane) dihydrochloride (AAPH)-damaged HDFs and the expression of Type I procollagen in HDFs were examined. HFEA caused the proliferation of HDFs with and without AAPH treatment and protected against AAPH damage to HDFs in a dose-dependent manner ($50-200{\mu}g/mL$). This implies that the antioxidant properties of the fractions depended on their proliferative and protective effects. The HFEA antioxidant fraction had significant effects and caused the dose-dependent expression of Type I procollagen, an important anti-wrinkle protein, in HDFs. In conclusion, antioxidant substances in H. fusiformis were found in the ethyl acetate fraction, and the resulting HFEA may have cosmetic applications.

• Journal of Radiation Protection and Research
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• v.44 no.1
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• pp.15-25
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• 2019

Background: Psammaplin A (PsA) is a radiosensitizer whereas its clinical application is hampered by poor bioavailability. This study aimed to synthesize novel radiosensitizers using PsA as the lead compound. Materials and Methods: Eight homodimeric disulfides were synthesized from corresponding acid and cystamine dihydrochloride in N-hydroxysuccinimide and dicyclohexylcarbodiimide coupling conditions. One monomeric thiol analog was obtained by reduction of homodimeric disulfide with dithiothreitol. Clonogenic assay was used to measure cell survival after irradiation and drug treatment in human lung cancer (A549) and glioblastoma (U373MG) cells. Results and Discussion: Using the PsA backbone, nine compounds were synthesized. Eight compounds showed variable cytotoxicity with 50% inhibitory concentrations ranging $16.14{\mu}M$ to $150.10{\mu}M$ (A549), and $13.25{\mu}M$ to $50.15{\mu}M$ (U373MG). Four and six compounds radiosensitized A549 and U373MG cells, respectively. Two compounds that radiosensitized both cell lines were tested for its inhibitory effects on DNMT1. One of them was shown to significantly inhibit DNMT1 activity. Conclusion: Novel compounds with radiosensitizing activity were synthesized. These compounds have a great potential to serve as a basis for the development of future radiosensitizers. Further investigation is warranted for their clinical application.

• International Journal of Oral Biology
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• v.46 no.1
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• pp.15-22
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• 2021

Alpha-lipoic acid (ALA) is a naturally occurring antioxidant and has been previously used to treat diabetes and cardiovascular disease. However, the autophagy effects of ALA against oxidative stress-induced dopaminergic neuronal cell injury remain unclear. The aim of this study was to investigate the role of ALA in autophagy and apoptosis against oxidative stress in the SH-SY5Y human dopaminergic neuronal cell line. We examined SH-SY5Y phenotypes using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay (cell viability/proliferation), 4′,6-diamidino-2-phenylindole dihydrochloride nuclear staining, Live/Dead cell assay, cellular reactive oxygen species (ROS) assay, immunoblotting, and immunocytochemistry. Our data showed ALA attenuated hydrogen peroxide (H2O2)-induced ROS generation and cell death. ALA effectively suppressed Bax up-regulation and Bcl-2 and Bcl-xL down-regulation. Furthermore, ALA increased the expression of the antioxidant enzyme, heme oxygenase-1. Moreover, the expression of Beclin-1 and LC-3 autophagy biomarkers was decreased by ALA in our cell model. Combined, these data suggest ALA protects human dopaminergic neuronal cells against H2O2-induced cell injury by inhibiting autophagy and apoptosis.

• The Korea Journal of Herbology
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• v.28 no.3
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• pp.25-31
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• 2013

Objectives : To develop a natural antioxidant and anti-hemolytic agents, we investigated the effects of ethanol extracts of Carpesii Fructus and Farfarae Flos. Methods : Aerial parts of Carpesii Fructus and Farfarae Flos were extracted with 80% ethanol. Antioxidant activity of Carpesii Fructus or Farfarae Flos extract was evaluated by employing three different assays, i.e., 1,1-diphenyl-2-picryl hydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzthiazoline-6-suphonic acid) diammonium (ABTS) scavenging and reducing power activities. Also, anti-hemolytic activity of Carpesii Fructus or Farfarae Flos extract was determined using [2,2'-azobis (2-methylpropionamidine) dihydrochloride (AAPH)]-induced hemolysis, glutathione (GSH) depletion and malondialdehyde (MDA) formation in normal rat red blood cells (RBC) or plasma. Results : The extracts obtained from Carpesii Fructus and Farfarae Flos dose-dependently increased the scavenging activity on DPPH- or ABTS-induced radicals and the reducing power activities. Carpesii Fructus and Farfarae Flos were similar to the scavenging activity and the reducing power of butylated hydroxy anisole effect at high concentration ($1,000{\mu}g/mL$). RBC oxidative hemolysis and plasma MDA formation induced by AAPH were significantly suppressed by the extracts of Carpesii Fructus and Farfarae Flos in a dose-dependent manner. Also, Carpesii Fructus and Farfarae Flos extracts prevented the depletion of cystosolic antioxidant GSH in RBCs. Carpesii Fructus generally had better than the free radical scavenging activity, the reducing power and anti-hemolytic effects of Farfarae Flos. Conclusions : These results suggest that Carpesii Fructus and Farfarae Flos may have value as the potential antioxidant and anti-hemolytic medicinal plant.

• Journal of Korean Medicine for Obesity Research
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• v.22 no.1
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• pp.1-10
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• 2022

Objectives: To investigate the protective effect of hesperidin and hesperetin against oxidative stress in 2,2'-azobis (2-aminopropane) dihydrochloride (AAPH)-induced liver toxicity in rats. Methods: Hesperidin or hesperetin (200 mg/kg/day, respectively) was orally administered for 7 days once daily in rats. Subsequently, AAPH (50 mg/kg/day) was administered intraperitoneally. Lipid peroxidation, nitric oxide production, catalase activity, and protein expressions of nuclear factor-kappa B (NF-κB) and inducible nitric oxide synthase (iNOS) in the liver tissues were measured. Results: Administration of hesperidin and hesperetin significantly decreased serum aspartate transaminase and alanine transaminase levels in AAPH-induced oxidative stress liver tissues compared with control group. Lipid peroxidation and nitric oxide (NO) production were also significantly reduced by hesperidin and hesperetin in AAPH-induced oxidative stress liver tissues. In particular, lipid peroxidation levels of hesperetin-administered group significantly decreased to 5.02 nmole/mg protein in oxidative stress rats. Hesperidin and hesperetin significantly increased antioxidant activity, such as that of catalase. Furthermore, administration of hesperidin and hesperetin substantially down-regulated the expression of NF-κB and iNOS in liver tissues. Administration of hesperidin reduced NO levels and iNOS expression more than in the hesperetin-administered group. Conclusions: Administration of hesperidin and hesperetin led to a reduction in AAPH-induced liver toxicity by regulating oxidative stress.

• Journal of Sasang Constitutional Medicine
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• v.11 no.2
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• pp.227-250
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• 1999

The free radical theory of aging was introduced in 1956 by Denham Harman. This aging theory proposed that normal aging results from random deleterious damage to tissues by free radical and supplying antioxidant lead to decrease oxidative damage, inhibit aging process. In this study, we investigated antioxidantive effects of four Korean constitutional prescriptions for 'Soum' constitution - Palmulgunjatang(Y1), Sipyimiguanjungtang(Y2), Osuyubujayijungtang(Y3) and Seungyangyikkibujatang(Y4). Antioxidative activity of this prescriptions was examined by 1,1-diphenyl-2-picrylhyrdazyl radicals, superoxide anion radicals, peroxyl radical, hydroxyl radical scavenging effects and erythrocyte hemolysis inhibitory effects. Y2 and Y3 were shown to have relatively high antioxidative activity on this methods. In additions, result of the cytoprotective effects of Korean constitutional prescriptions agianst 2,2'-azobis(amidinopropane) dihydrochloride (AAPH), a free radical initiator, induced cytotoxcity in human hepatoblastoma cell line was similarly obtained. On the basis of this result, we assayed the antioxidative effects of Y2 and Y3 on experimental oxidative damage, induced in mouse by 100mg/kg AAPH. Male ICR mouse were given oral administration of 500mg/kg Y2 and Y3 for 4 weeks. Thiobarbuturic acid reactive substance (TBARS) and protein degradation level in liver, plasma and brain as index of oxidative damage were decreased and thiol compound, total antioxidant status in plasma were increased by Y2 administration. But, Y3 injected group was decreased only protein degradation level in brain. Also, glutathione, a potent water-soluble endogenous antioxidant, concentration was increased by Y2 and Y3 administration in liver and brain. However, superoxide dismutase and catalase activity as a major antioxidative enzyme in vivo were not shown change by Y2 and Y3 administration. On the basis of these result, Y2 have an antioxidative effects on both water-soluble fraction and lipid-solube fraction in cell and tissues. But, Y3 has a lower antioxidative effects on lipid-soluble fraction than Y2 in cell and tissues. These results suggest that Y2 has a antioxidative effects by protect the tissue against oxygen free radical mediated oxidative damage and Y3 has a limited antioxidaitve effects on water-soluble fraction in vivo. Therefore, we make report that Y2 is more effective prescriptions for anti-aging or therapeutics of diseases.

• International Journal of Oral Biology
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• v.30 no.2
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• pp.47-57
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• 2005

Leptin, the product of the obese gene, is a circulating hormone secreted primarily from adipocytes. Several results suggest that leptin is important mediators of bone metabolism. The present study was undertaken to determine the effects of leptin on anti-osteoclastogenesis using murine precursors cultured on Ca-P coated plates and on the production of osteoprotegerin (OPG) in osteoblastic cells. Additionally, this study examined the possible involvement of prostaglandin $E_2\;(PGE_2)$/protein kinase C (PKC)-mediated signals on the effect of leptin on anti-osteoclastogenesis to various culture systems of osteoclast precursors. Osteoclast generation was determined by counting tartrate-resistant acid phosphatase positive [TRAP (+)] multinucleated cells (MNCs). Osteoclastic activity was determined by measuring area of resorption pits formed by osteoclasts on Ca-P coated plate. The number of 1,25-dihydroxycholecalciferol $(1,25[OH]_2D_3)$- or $PGE_2$-induced TRAP (+) MNCs in the mouse bone marrow cell culture decreased significantly after treatment with leptin. The number of receptor activator of NF-kB ligand (RANKL)-induced TRAP (+) MNCs in M-CSF dependent bone marrow macrophage (MDBM) cell or RAW264.7 cell culture decreased significantly with leptin treatment. Indomethacin inhibited osteoclast generation induced by $1,25[OH]_2D_3$ and dexamethasone, however, no significant differences were found in the leptin treated group when compared to the corresponding indomethacin group. Phorbol 12-myristate 13-acetate (PMA), a PKC activator, inhibited osteoclast generation induced by $1,25[OH]_2D_3$. The number of TRAP (+) MNCs decreased significantly with treatment by PMA at concentrations of 0.01 and $0.1{\mu}M$ in culture. Leptin inhibited PMA-mediated osteoclast generation. Isoquinoline-5-sulfonic 2-methyl-1-piperazide dihydrochloride (H7) had no effect on osteoclast generation induced by $1,25[OH]_2D_3$. Cell culture treatment with leptin resulted in no significant differences in osteoclast generation compared to the corresponding H7 group. Indomethacin showed no significant effect on TRAP (+) MNCs formation from the RAW264.7 cell line. PMA inhibited TRAP (+) MNCs formation induced by RANKL in the RAW264.7 cell culture. H7 had no effect on osteoclast generation from the RAW264.7 cell line. There was no difference compared with the corresponding control group after treatment with leptin. $1,25[OH]_2D_3$- or $PGE_2$-induced osteoclastic activity decreased significantly with leptin treatment at a concentration of 100 ng/ml in mouse bone marrow cell culture. Indomethacin, PMA, and H7 significantly inhibited osteoclastic activity induced by $1,25[OH]_2D_3$ in mouse bone marrow cell culture. No significant differences were found between the leptin treated group and the corresponding control group. The secretion of OPG, a substance known to inhibit osteoclast formation, was detected from the osteoblasts. Treatment by leptin resulted in significant increases in OPG secretion by osteoblastic cells. Taken these results, leptin may be an important regulatory cytokines within the bone marrow microenvironment.