• The Journal of Pediatrics of Korean Medicine
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• v.27 no.4
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• pp.108-121
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• 2013

Objective This study was designed to investigate the effects of Gamiygin-tang (GY) extract (GYE) and fermented solution (GYF) on body weight, serum lipid level and adipocyte differentiation in high fat diet-fed obese mice. Materials and Methods High fat diet-fed obese mice and 3T3-L1 adipocytes mice were treated with GYE and GYF and obesity related markers were assessed. A cytotoxicity assay was carried out by MTS assay. Inhibitory effects of GYE and GYF on adipocyte differentiation were carried out by Oil Red O staining. The effects of GYE and GYF on the expression of adipocyte differentiation regulatory factors, peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) and CCAAT/enhancer binding protein alpha (CEBP-${\alpha}$) were measured by real-time reverse transcriptase-polymerase chain reaction. The effects of GYE and GYF on the expression of adipocyte differentiation regulatory factors were also determined in relation to protein production/protein levels by western blotting. The anti obesity effects of GYE and GYF were measured in high fat-diet induced obese mice. Various factors were measured from the serum of the high fat-diet mice. Results Though GYE did not show toxicity at the concentration of 1mg/ml, GYF showed toxicity at the concentration of 1mg/ml. The GYE at 0.1 and 1mg/ml inhibited the differentiation of 3T3-L1 cells, and the GYF also inhibited the differentiation of 3T3-L1 cells. The effect of GYE on adipocyte differentiation factors including PPAR-${\gamma}$ and CEBP-${\alpha}$ was investigated and compared to the corresponding concentration levels of GYF. GYE and GYF both suppressed the RNA and protein levels of adipocyte differentiation factors. In the animal test both GYE and GYF reduced weight gain. GYE and GYF reduced blood cholesterol, TG and LDL levels. GYF better reduced blood cholesterol levels. Conclusions These results demonstrate that GYE and GYF exerts anti-obesity effect in 3T3-L1 cells and obese mice induced by high-fat diet.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.42 no.5
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• pp.434-441
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• 2009

This study was conducted to compare the biological activities of 7 melania snails from the family Pleuroceridae (Semisulcospira coreana, Koreanomelania nodifila, Semisulcospira forticosta, Koreoleptoxis globus ovalis, Semisulcospira libertina, Semisulcospira tegulata and Semisulcospira gottschei) in Korea. Among the 7 species, S. coreana, Korean. nodifila, S. forticosta and S. gottschei showed over 80% cytotoxicities on three cancer cell lines (SNU-1, A549 and Hep 3B) compared to the non-treatment, whereas S. libertina and S. tegulata showed almost no growth inhibition activities on the same cancer cell lines. In relation to ACE inhibition activity, only S. coreana, Korean. nodifila, and S. forticosta showed over 60% ACE inhibition activities, whereas other melania snails exhibited inhibition activities of lower than 25%. DPPH radical scavenging activities were also determined, and used to categories melania snails into three groups based on Duncan's multiple range test at P<0.05. The amount of TNF-${\alpha}$ produced by in vitro mouse peritoneal macrophage was determined according to bioactivity on L-929 cells. Three melania snails, S. coreana, Korean. nodifila and S. gottschei, exhibited 95.2%, 89.7% and 93.7% cell death(%) on L-929 cells, respectively. Glucose-6-phosphate dehydrogenase inhibitory activity was also obtained in the extract of S. coreana (31.9%) and Korean. nodifila (28.1%), showing that these extracts can be used as supplemental dietary health foods. In conclusion, we believe that the extracts of melania snails should be given due consideration in functional health food development.

• Korean Journal of Oriental Medicine
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• v.18 no.3
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• pp.103-110
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• 2012

Objectives : Fire needling has been applied as the treatment for various diseases and been getting much attention from Oriental medicine due to its excellent effectiveness as the results of clinical studies have reported. However, the research findings on the safety of treatment method, materials for the Fire needling needle materials and the possibility of burn injury during the procedure are still insufficient. Methods : A thermo imaging camera was used to confirm the temperature distribution on acupuncture needle and the treatment area during the fire needling therapy. Then the degree of thermal injury was observed by H&E stain and TUNEL assay. In addition, in order to assess the safety of acupuncture materials, we conducted MTT assay using a L6 cell line. Results : The average temperature of the skin surface was observed at $47{\sim}51^{\circ}C$ after classic fire needling and $30^{\circ}C$ after warming fire needling. Warming fire needling therapy does not induce a burn on the tissue and a third degree burn was observed locally in the muscle and skin layers after classic fire needling treatment. This confirms that hwa-acupuncture therapies do not cause major burns. According to the safety assessment test result, no cytotoxicity was detected in the warming fire needling materials. This confirms the safety of the acupuncture materials Conclusions : Various research results on the biological safety of fire needling. Since fire needling therapy induces a burn locally without leaving any scar, and as other results indicate, it is considered a safe treatment method.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.18 no.3
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• pp.513-517
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• 2017

The purpose of this study was to investigate the antioxidative and anti-inflammatory activities of ethanol extracts from Perilla frutescens var. acuta by varying the concentration of ethanol at 30, 50, 70, and 90% to utilize the effective extract of Perilla frutescens as a cosmetic and pharmaceutical material. In the DPPH antioxidant activity test, the 70% ethanol extract showed the highest activity with an IC50 of 680.98ppm. ABTS showed a high activity in the 50% ethanol extract and the 70% ethanol extract with an IC50 of 646.94 and 661.94 ppm, respectively. Each ethanol extract showed antioxidant activity at a certain concentration (100-10000 ppm), but did not show any significant relationship with the ethanol extract concentration. In RAW 264.7 macrophages induced by LPS, each of the ethanol extracts showed reduced NO production in all extracts, and more than 50% ethanol extract (10000ppm) inhibited nitric oxide formation by 85% or more. In particular, the 70% ethanol extract showed 90% or more nitric oxide production inhibition. In addition, the MTT assay showed no cytotoxicity at all concentrations (1250-10000 ppm) of each extract. In this study, the ethanol extract of Perilla frutescens var. acuta has antioxidant activity and anti-inflammatory activity that is dependent on the concentration at each extraction concentration.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.31 no.3
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• pp.39-49
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• 2018

Objectives : This study is designed to clarify whitening, anti-inflammatory effect of fractions extracted from the mixture of Phaseolus radiatus L. and ethanol. Methods : In this experiment, we were intended to reveal whitening, anti-inflammatory effect of fractions extracted from the mixture of Phaseolus radiatus L. and ethanol. The whitening activity was confirmed by UV blocking activity, tyrosinase inhibiting activity, and melanin formation inhibiting activity. Anti-inflammatory activity is confirmed by measurement of cytotoxicity level by MTT assay and measurement of Cytokine expression, which is the main mediator of inflammation reaction. Results : As a results, overall activity was high in the ethyl acetate fraction. Tyrosinase inhibitory activity was less than 20% at all concentrations, but the activity to inhibit melanin self-production was higher than that of ethyl acetate fraction at $32.19{\pm}2.79%$ at $100{\mu}g/m{\ell}$. And ethyl acetate fraction had a relatively high UV blocking activity. In the anti-inflammatory test, the concentration-dependent activity was shown, and the chloroform and ethyl acetate fractions showed significant NO production inhibitory activity. Cytokine expression was superior to that of the final stage of B cell differentiation, and cell viability was over 80% except for the chloroform fraction at the concentration of $200{\mu}g/m{\ell}$. Conclusions : The results of this experiment confirmed the whitening effect and anti-inflammatory effect of Phaseolus radiatus L.'s extracts and fractions and report the possibility of application as external medicine.

• Korean Journal of Food Science and Technology
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• v.31 no.4
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• pp.1065-1070
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• 1999

This study was performed to determine the antimutagenic and cytotoxic effect of Aster scaber root ethanol extract on Salmonella typhymurium TA98, TA100 and cancer cell lines using Ames test and cytotoxicity assay, respectively. Cancer cell lines include chronic myelogenous leukemia(K562), human gastric carcinoma(KATOIII), human hepatocellular carcinoma(Hep3B) and human breast adenocarcinoma(MCF-7). Futher fractionations with hexane, chloroform, ethyl acetate, butanol and water from ethanol extract of Aster scaber root were performed to obtain effective fraction. Ethanol extract and ethyl acetate fraction showed 79% and 82% inhibitory effect on the mutagenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) against TA100, while 48% and 60% inhibition was observed on the mutagenesis induced by 4-nitroquinoline-l-oxide(4NQO) against TA98. In the meanwhile, ethyl acetate fraction showed 78% and 85% inhibitory effect on the mutagenesis induced by benzo(${\alpha}$)pyrene[B(${\alpha}$)P] against TA98 and TA100, respectively, while 83% inhibition was observed on the mutagenesis induced by 3-amino-l,4-dimethyl-5H-pyrido(4,3-b) indole(Trp-P-1) against TA98. Ethyl acetate fraction (0.125 mg/ml) showed the strongest cytotoxic effect against K562, KATOIII, Hep3B and MCF-7 at the same concentration compared to those of other fractions. Ethanol extract and water fraction showed the least inhibitory effect.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.9
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• pp.1357-1362
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• 2014

This study compared the antioxidant effects of two kinds of black raspberry extract, obtained from fruits of Rubus coreanus and Rubus occidentalis, which can be found in Korea. The fruits of R. coreanus and R. occidentalis were each extracted with 0%, 25%, 50%, 75%, and 100% ethanol (EtOH). Among the extracts of these two varieties, 50% EtOH extract of R. occidentalis showed the highest contents of total polyphenols ($46.96{\pm}2.78mg/g$) and flavonoid compounds ($11.77{\pm}0.81mg/g$). The 50% EtOH extract of R. occidentalis showed the highest antioxidant activity ($84.77{\pm}0.97%$) in terms of DPPH radical scavenging activity. On the contrary, 25% EtOH extract of R. occidentalis showed the best antioxidant activity ($29.65{\pm}2.41%$) in terms of ABTS radical scavenging activity. In the results of ferric reducing antioxidant power (FRAP) assay, 50% EtOH extract of R. occidentalis showed the highest antioxidant activity ($0.49{\pm}0.02%$). In the cytotoxicity test stimulated with $H_2O_2$, the extracts of 75% and 100% EtOH from R. occidentalis showed the highest cell viability ($93.54{\pm}3.37%$ and $97.19{\pm}0.74%$, respectively). According to our results, extracts of R. occidentalis showed higher antioxidant activities than extracts of R. coreanus. Especially, total polyphenol and flavonoid contents of R. occidentalis showed the highest significant correlation with FRAP by Pearson's correlation (P=0.005 and P=0.013, respectively).

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.2
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• pp.397-404
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• 2009

For standardization of LMK02 quality, Ginsenoside Rg3 of Red Ginseng and Decursin of Angelica gigas Nakai in the constituents of LMK02 were estimated as indicative components. From LMK02 water extract, has been used in vitro test for its beneficial effects on neuronal survival and neuroprotective functions, particularly in connection with APP-related dementias and Alzheimer's disease (AD). $A{\beta}$ oligomer derived from proteolytic processing of the ${\beta}$-amyloid precursor protein (APP), including the amyloid-${\beta}$ peptide ($A{\beta}$), play a critical role in the pathogenesis of Alzheimer's dementia. We determined that oligomer amyloid-${\beta}$ ($A{\beta}$) have a profound attenuation in the increase in rat hippocampus H19-7 cells from. Experimental evidence indicates that LMK02 protects against neuronal damage from cells, but its cellular and molecular mechanisms remain unknown. Using a hippocampus cell line on $A{\beta}$ oligomer-induced neuronal cytotoxicity, we demonstrated that LMK02 inhibits formation of $A{\beta}$ oligomer, which are the behavior, and possibly causative, feature of AD. In the Red Ginseng, the average amounts of Ginsenoside Rg3 were $47.04{\mu}g/g$ and $42.3{\mu}g/g$, 90 % of its weight were set as a standard value. And, in the Angelica gigas Nakai, the average amounts of Decursin were 2.71 mg/g and 2.44mg/g, 90 % of its weight were also set as a standard value. The attenuated $A{\beta}$ oligomer in the presence of LMK02 was observed in the conditioned medium of this $A{\beta}$ oligomer-induced cells under in vitro. In the cells, LMK02 significantly activated antiapoptosis and decreased the production of ROS. These results suggest that neuronal damage in AD might be due to two factors: a direct $A{\beta}$ oligomer toxicity and multiple cellular and molecular neuroprotective mechanisms, including attenuation of apoptosis and direct inhibition of $A{\beta}$ oligomer, underlie the neuroprotective effects of LMK02 treatment.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.12
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• pp.6191-6196
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• 2012

Aims and Background: Traditional chemotherapy strategies for human leukemia commonly use drugs based on cytotoxicity to eradicate cancer cells. One predicament is that substantial damage to normal tissues is likely to occur in the course of standard treatments. Obviously, it is urgent to explore therapies that can effectively eliminate malignant cells without affecting normal cells. Our previous studies indicated that ginsenoside $Rg_1$ ($Rg_1$), a major active pharmacological ingredient of ginseng, could delay normal hematopoietic stem cell senescence. However, whether $Rg_1$ can induce cancer cell senescence is still unclear. Methods: In the current study, human leukemia K562 cells were subjected to $Rg_1$ exposure. The optimal drug concentration and duration with K562 cells was obtained by MTT colorimetric test. Effects of $Rg_1$ on cell cycle were analyzed using flow cytometry and by SA-${\beta}$-Gal staining. Colony-forming ability was measured by colony-assay. Telomere lengths were assessed by Southern blotting and expression of senescence-associated proteins P21, P16 and RB by Western blotting. Ultrastructural morphology changes were observed by transmission electron microscopy. Results: K562 cells demonstrated a maximum proliferation inhibition rate with an $Rg_1$ concentration of $20{\mu}\;mol{\cdot}L^{-1}$ for 48h, the cells exhibiting dramatic morphological alterations including an enlarged and flat cellular morphology, larger mitochondria and increased number of lysosomes. Senescence associated-${\beta}$-galactosidase (SA-${\beta}$-Gal) activity was increased. K562 cells also had decreased ability for colony formation, and shortened telomere length as well as reduction of proliferating potential and arrestin $G_2$/M phase after $Rg_1$ interaction. The senescence associated proteins P21, P16 and RB were significantly up-regulated. Conclusion: Ginsenoside $Rg_1$ can induce a state of senescence in human leukemia K562 cells, which is associated with p21-Rb and p16-Rb pathways.

• Korean Journal of Plant Resources
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• v.29 no.5
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• pp.525-531
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• 2016

Cynanchum wilfordii Radix (CWR), Arctium lappa L (ALL), and Dioscorea opposite (DO) have been known to improve blood lipid profile, blood pressure, and inflammation. To find the optimal combination ratio of CWR, ALL, and DO in terms of vascular health improvement, we compared the effects of various combinations on gene expression of Vascular cell adhesion protein 1 (VCAM-1) in human aortic smooth muscle cells (HASMC). VCAM-1 mediates endothelial leukocyte adhesion and is upregulated in atherosclerosis. Cells was stimulated by TNF-α (10 ng/㎖, 2h) and treated with various combinations for 24 h. A combination (CADM5, CWR:ALL:DO = 2:1:1) showed the strongest suppression of VCAM-1 so that CADM5 was chosen for further experiments. We performed cell viability test with CADM5 (0, 3.125, 12.5, 25, 50, and 100 ㎍/㎖) and no cytotoxicity was found. We also investigated the effect of CADM5 on protein expression of VCAM-1, ICAM-1, Nrf-2, and HO-1 using western blotting. We found that CADM5 diminished the expression of VCAM-1 and increased the expression of Nrf-2 and HO-1. Therefore, we concluded that CADM5 (CWR:ALL:DO = 2:1:1) effectively improves vascular health by regulating the expression of VCAM-1.