• Proceedings of the Korean Society of Crop Science Conference
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• 2003.05a
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• pp.41-41
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• 2003

• Herbal Formula Science
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• v.26 no.3
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• pp.183-193
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• 2018

Objectives : In Traditional Korean Medicine, Epimedium koreanum Nakai has diverse pharmacological activities to treat impotence, forgetfulness, cataract and exophthalmos. Present study investigated anti-fibrogenic effects of E. koreanum water extract (EKE) in hepatic stellate cells (HSCs). Methods : To study anti-fibrogenic effects of EKE, LX-2 cells, a human immortalized HSCs, were pre-treated with $3-300{\mu}g/mL$ of EKE, and then subsequently exposed to 5 ng/mL of transforming growth $factor-{\beta}1$ ($TGF-{\beta}1$). Expression level of ${\alpha}-smooth$ muscle actin was determined by immunoblot analysis. Phosphorylation of Smad, transactivation of Smad, and expression of plasminogen activator inhibitor-1 (PAI-1) were monitored to investigate the effect of EKE on $TGF-{\beta}1-mediated$ signaling pathway. Results : Up to $100{\mu}g/mL$, EKE did not show any cytotoxicity on LX-2 cells. Pre-treatment of EKE ($100{\mu}g/mL$) significantly inhibited ${\alpha}-smooth$ muscle actin expression induced by $TGF-{\beta}1$. In addition, EKE significantly decreased Smad2 and Smad3 phosphorylations, Smad binding element-driven luciferase activity and PAI-1 expression by $TGF-{\beta}1$. Of three flavonoid compounds found in EKE, only quercertin ($30{\mu}M$) attenuated $TGF-{\beta}1-mediated$ PAI-1 expression. Conclusion : These results suggest that EKE has an ability to suppress fibrogenic process in HSCs via inhibition of $TGF-{\beta}1/Smad$ signaling pathway.

• Journal of Veterinary Clinics
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• v.28 no.4
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• pp.357-364
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• 2011

To evaluate hepatoprotective effect of Epimedium Koreanum nakai (EKN) on liver-damaged animal model, rats were intoxicated with carbon tetrachloride (1 ml/kg) for 9 weeks orally. Liver-damaged rats were divided into 2 groups: liver-damaged control (LDC) group and EKN group were administered vehicle (saline), EKN extract per os for 4 weeks respectively. Normal control (NC) group was administered saline as the same process of LDC group. The weights of prostate (absolute), testis (relative), epididymis (relative) and packed cell volume (PCV), mean corpuscular volume (MCV) of EKN group significantly (P < 0.05) increased compared with LDC group. But Mean corpuscular hemoglobin (MCH), mean corpusulcar hemoglobin concentration (MCHC) and aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) were significantly (P < 0.01, P < 0.05) decreased. Fibrotic regions in hepatic parenchyma of EKN group significantly (P < 0.01) decreased compared with LDC group and mean diameters of hepatic lobules significantly (P < 0.01) increased. Percentages of degenerative kidney regions and number of degenerative kidney tubules, number of vasodilated atrophic glomerulus of EKN group was significantly (P < 0.01, P < 0.05) decreased compared with LDC group. Number of atrophic seminiferous tubules and epididymal tubules showing oligospermatozoa of EKN group were significantly (P < 0.01) decreased compared with LDC group. In conclusion, EKN extract has a favorable effect on the $CCl_4$-induced liver damage.

• Journal of Veterinary Clinics
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• v.27 no.2
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• pp.163-169
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• 2010

To evaluate of oral toxicity of Epimedium koreanum nakai (EKN) water extract, three doses of EKN water extract (10, 100 and 1000 mg/kg/day) were administered to healthy Sprague-Dawley rats for 4 weeks. The results showed that there are no abnormal signs during the experimental period. The weights of testis and epididymis were increased dose-dependently, but seminal vesicle was decreased compared with control group. In hematology and serum chemistry, changes were observed within the normal ranges. In histological finding, slight to moderated changes were found but those were commonly or rarely encountered in the normal rats. These results suggest that no observed adverse effect level (NOAEL) of the oral application of EKN was considered to be more than 1000 mg/kg in SD rats under the conditions employed in this study.

• Journal of Physiology & Pathology in Korean Medicine
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• v.27 no.6
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• pp.730-737
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• 2013

The purpose of this study is to investigate vasorelaxant effect of Epimedium koreanum(EK) extract on rabbit carotid artery. In this study, to determine vasorelaxant effect of EK extract on rabbit carotid artery, arterial rings with intact or damaged endothelium were used for experiment using organ bath, and were contracted by norepinephrine(NE). After being contracted, arterial rings were treated with EK extract in a dose-dependent manner To study its mechanism, the contracted arterial rings induced by NE were pretreated with indomethacin(IM), $N_{\omega}$-nitro-L-arginine(L-NNA), methylene blue(MB) or tetraethylammonium chloride(TEA) and 0.1 $mg/m{\ell}$ EK extract was added. To analyze the effect of the EK extract on influx of extracellular calcium chloride($Ca^{2+}$) in rabbit carotid artery, in $Ca^{2+}$-free krebs solution, krebs solution containing 1 mM $Ca^{2+}$ was infused into the contracted arterial ring by NE after pretreatment of EK extract. To measure the cytotoxicity of the EK extract, cell viability of human umbilical vein endothelial cell(HUVEC) was measured by MTT assay, and nitric oxide(NO) was measured by Griess reagent. The EK extract significantly was relaxed the arterial ring with intact endothelium contracted by NE, but the vasorelaxant effect of the EK extract was inhibited in the arterial rings with damaged endothelium. The vasorelaxant effect of the EK extract was not different between the IM-pretreatedand and non-treated arterial rings. The vasorelaxant effect of EK extract were significantly inhibited, when arterial rings were pretreated with L-NNA, TEA, MB. And in $Ca^{2+}$-free krebs solution, increasing of arterial contraction by $Ca^{2+}$ addition were also inhibited by the treatment of EK, but not significant. The treatment of EK extract was increased NO concentration in HUVEC. This study suggested that the vasorelaxant effect of EK extract would be related with EDHF and NO production and increasing of cyclic GMP.

• Journal of Physiology & Pathology in Korean Medicine
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• v.28 no.2
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• pp.169-177
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• 2014

This study aimed to investigate the relaxation effects and its underlying mechanisms of Epimedium koreanum Nakai(EK) in phenylephrine(PE) treated isolated rabbit corpus cavernosum smooth muscle. The dose-dependent relaxation responses of phenylephrine(PE, $1{\times}10^{-6}M$)-precontracted strips to EK at $0.01-3.0mg/m{\ell}$ were measured and also observed after endothelial denudation using organ bath. To analyze the underlying mechanisms of EK-induced relaxation, $N{\omega}$-nitro-L-arginine(L-NNA), methylene blue(MB), tetraethylammonium chloride(TEA), indomethacin(IM) were pretreated before EK extract infused into precontracted strips induced by PE. To investigate cytotoxic activity and nitric oxide(NO) concentration of EK extract on EA.hy926 cells, mitochondrial dehydrogenase activity(MTT) assay and nitric oxide detection kit were used. The cavernous strips were significantly relaxed by EK extract at $0.3mg/m{\ell}$, $1.0mg/m{\ell}$, $3.0mg/m{\ell}$ and the relaxation responses of PE-precontracted strips denuded endothelium also inhibited in comparison with intact endothelium. The pretreatment of L-NNA, MB, TEA reduced EK extract-induced endothelium-dependent relaxation, but the pretreatment of IM didn't affect EK extract-induced endothelium-dependent relaxation. When EK extract was applicated on EA.hy926 cells, the NO concentration was increased. Our findings have shown that EK extract exerts a relaxing effect on corpus cavernosum in part by suppressing influx of extracellular $Ca^{2+}$ through activating the NO-cGMP system.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.35 no.4
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• pp.265-270
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• 2009

To develop a natural stimulating agent for hair melanogenesis, we investigated the stimulatory effects of Epimedium koreanum Nakai extract (EKNE) on melanogenesis. EKNE both increased melanin content in B16 melanoma cells up to about 104 % and intracellular tyrosinase activity up to about 95 % at a concentration of $50\;{\mu}g/mL$ without cell cytotoxicity (below $100\;{\mu}g/mL$). From the result of western blot, we could find that EKNE dose-dependently induce the expression of TRP-2. Also, EKNE increased melanogenesis up to about 25 % at a concentration of 5 % (w/v) in back hair of C3H/Hej mouse. These results suggest that EKNE has an effect to stimulate melanin formation through the induction of tyrosinase activity and TRP-2 expression which are involved in melanin synthesis in melanocyte. Therefore, we suggest that EKNE could be used as a useful melanogenesis stimulator for development of natural hair cosmetics.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.4
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• pp.732-736
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• 2000

The purpose of this study was to investigate the extraction method of phenolic compounds and antioxidant activities of the aerial part of Epimedium koreanum NAKAI (EKN). The antioxidant activities of EKN were tested with by hydrogen donating ability, inhibition of lipid peroxidation, and inhibition of LDL oxidation. The most suitable conditions for the extraction of phenolic compounds from EKN were to use 60% ethanol by 3 times, and the yield of extract (dry basis) was 22%. In the extraction efficiency of phenolic compounds, 60% ethanol as extracting solvent was superior to water. Sixty% ethanol extract of EKN was found to have an ability of hydrogen donating to DPPH. MDA determination showed the 95% inhibitory effect against linoleic acid oxidation by the addition of 700 ppm EKN extract. Also, about 70% of LDL oxidation was inhibited by the addition of 500 ppm.

• International Journal of Oral Biology
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• v.42 no.4
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• pp.163-168
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• 2017

Osteoporosis is a metabolic bone disease that is characterized by low bone mass resulting from an increase in bone resorption relative to bone formation. The most current therapies for osteoporosis have focused on inhibiting bone resorption by osteoclasts. The purpose of this study is to develop new anabolic agents for treatment of osteoporosis that have fewer risks compared to conventional therapies. We searched the natural products that were derived from the traditional Asian medicines which have been used for treatment of bone related diseases. Icaritin is a flavonoid glycoside derived from the herb Epimedium which has beneficial effects on bone formation. To determine the effect of icaritin on bone formation, we examined the effect of icaritin on MC3T3-E1 cell proliferation and differentiation. For determining the effects of icaritin on proliferation, we performed the MTT assay using MC3T3-E1 cells. To evaluate whether icaritin could promote the osteogenic differentiation of MC3T3-E1 cells, alkaline phosphatase (ALP) activity and mRNA expressions of Runx2, osteocalcin (OCN), RANKL, and osteoprotegerin (OPG) were determined. Icaritin increased MC3T3-E1 cell proliferation. Icaritin increased the ALP activity of MC3T3-E1 cells on 72 hour culture in osteogenic media. mRNA expression of Runx2 was increased after 24 hour culture with icaritin. mRNA expression of osteocalcin was increased after 72 hour culture with icaritin. In addition, icaritin increased the mRNA expressions of OPG and RANKL. However, icaritin increased the mRNA expression of OPG much more than that of RANKL, and then, it increased the OPG/RANKL ratio. These results suggest that icaritin promotes osteogenic differentiation of osteoblasts and decreases osteoclast formation regulated by osteoblasts.

• Journal of Food Hygiene and Safety
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• v.22 no.4
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• pp.359-364
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• 2007

This study was conducted to determine the optimum condition of extraction and antioxidant activity of ethanol extracts of Epimedium koreanum Nakai based on the icariin quantity. Also, further organic solvent fractions of hexane, chloroform, ethyl acetate and butanol were obtained from the ethanol extract of Epimedium koreanum Nakai at different temperatures. Total ethanol extraction yield of wild grape seed ranged from 11.8% to 39.3% depending on the concentration of icariin as well as different ethanol concentration, extraction temperature and time condition. Among different extraction temperatures and time, the highest extraction yield of 39.3% was obtained at 70% ethanol for 3 hour at $50^{\circ}C$ in the sample containing the 0.596% icariin (KE9412). In the meantime, the strongest free radical scavenging effect $(RC_{50})$ with $18.9{\mu}g/mL$ in the KE9412 sample was observed in 70% ethanol extract of Epimedium koreanum Nakai extracted for 7 hour at $70^{\circ}C$, while $RC_{50}$ with $35.2{\mu}g/mL$ was observed in the KE9405 (0.20% icariin content) sample at same condition. Also, antioxidant activity of ethanol extracts of Epimedium koreanum Nakai increased as icariin concentration increased. Among each fraction obtained from organic solvents, butanol fraction was found to have the strongest $RC_{50}\;(39.2{\mu}g/mL)$ and followed by ethylacetate $(49.0{\mu}g/mL)$, water $(118.8{\mu}g/mL)$, ethylacetate $(119.8{\mu}g/mL)$, and chloroform fraction $(138.5{\mu}g/mL)$ respectively.