• 대한임상검사과학회지
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• 제48권3호
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• pp.176-182
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• 2016

대식세포에서 염증반응이 진행되면, interleukin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) 등의 cytokine들이 발현되어 inducible nitric oxide synthase (iNOS), prostaglandin E2(PGE2) 등의 염증유발인자가 생성된다. 오가피, 우슬, 두충 각각의 추출물이 어느 정도의 항 염증 효능을 보이며 어떤 pro-inflammatory cytokine의 발현을 억제하는지에 대한 연구를 진행하였다. 오가피, 우슬, 두충은 물 추출하고 동결 건조시켰다. 각각의 추출물의 구성 성분들이 잘 추출되었는지 확인하기 위하여 지표물질인 acanthoside D, 20-hydroxyecdysone, pinoresinol diglucoside를 HPLC로 분석하였다. 항 염증 효능을 확인하기 위하여 lipopolysaccharide (LPS)로 RAW 264.7 세포주를 자극하여 염증 반응을 일으킨 상태에서 각각의 추출물을 농도 별로 처리하고 NO assay를 통해 항 염증 효능을 확인하였으며 real time PCR로 pro-inflammatory cytokine들의 발현량을 측정하였다. 결과적으로 각각의 추출물은 지표성분들이 검출되었으며 오가피와 우슬이 두충보다 NO assay에서 높은 활성을 보였다. Cytokine 발현량 측정에서는 오가피와 우슬은 iNOS와 IL-6의 발현을 억제하였고, 우슬은 TNF-${\alpha}$의 발현을 억제하였다. 우리나라는 전통적으로 약재를 조합하여 처방하여 왔다. 본 연구는 관절염에 전통적으로 사용해 오던 약재들이 어떤 기전에 의하여 항 염증 반응을 보이는지 확인하고 이들을 조합하여 사용하였을 때 어떤 근거에 의하여 시너지 효능을 보이는지 확인하였다.

• Journal of Acupuncture Research
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• 제32권2호
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• pp.1-9
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• 2015

Objectives : The present study was an evaluation and standardization of herbal components in order to establish the efficacy and safety of Shinbaro pharmacopuncture. Methods : Among the raw materials of Shinbaro pharmacopuncture, the components Cibotii Rhizoma, Eucommiae Cortex, and Ledebouriellae Radix were assessed through ingredient verification experiments using thin-layer chromatography(TLC) and ultraviolet rays(UV) lamps. In addition, we standardized Acanthopanacis Cortex and Achyranthis Radix through validation using high performance liquid chromatograph-diode array detector(HPLC-DAD). Results : As result appeared a blue-white fluorescence under ultraviolet rays; changed to dark green after adding 1 % ferric chloride solution(due to Cibotii Rhizoma), and presented a yellow-green fluorescence when mixed with an ethyl ether under UV lamps by way of the ethyl ether layer, confirming Eucommiae Cortex. Ledebouriellae Radix was confirmed as dark brown spots at Rf values of 0.56 and 0.71 using TLC. Additionally, Acanthopanacis Cortex and Achyranthis Radix HPLC test results showed that linearity was $R^2{\geq}0.99$, and detection limit and quantitation limit were 0.23 to $1.29{\mu}g/mL$, and 0.71 to $3.90{\mu}g/mL$, respectively. Furthermore, precision and accuracy were confirmed to have relative standard deviation(RSD) values of 0.10 to 1.89 % and 96.19 to 103.72 %, respectively. Shinbaro pharmacopuncture did not have any overlapping or interference from other peaks in detection under the abovementioned analysis conditions. Conclusions : In conclusion, we confirmed that maintenance of Shinbaro pharmacopuncture validity was possible by means of quality control of Cibotii Rhizoma, Eucommiae Cortex, and Ledebouriellae Radix through ingredient identification and Acanthopanacis Cortex and Achyranthis Radix through high performance liquid chromatograph(HPLC) analysis. Further, we hope to contribute to the development strategy of herbal industry acupuncture.

• 동아시아식생활학회지
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• 제23권1호
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• pp.39-43
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• 2013

Carthamus tinctorius L. and Eucommia umoides Oliver are often used in traditional herbal medicines for reducing damage to the liver, kidney, bone and muscle. In the present study, we investigated cell viability and alkaline phosphatase activity in the human osteoblast-like MG-63 cell line with methanol extracts of Carthami Semen (CS) and Eucommiae Cortex (EC) alone or in a mixture (CS+EC). Osteoblast cell viability was evaluated using the MTS assay and alkaline phosphatase activity assays. The cell viability and alkaline phosphatase activity significantly increased in MG-63 osteoblast cells treated with the CS+EC mixture. These findings suggest the CS+EC mixture may have beneficial effects on bone health through the proliferation of osteoblast cells.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.596-599
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• 2000

30 kinds of natural products were considered for developing natural antioxidants by improved D.O. analyzing method including simple calculation of Area Under Curve. Several natural products such as Cimicifuge Rhizoma, Epimedii Herba, Atractylodis Rhizoma Alba, Acori Graminei Rhizoma, Mori Cortex Radicis, Aurantii Nobilis Pericarpium were bore nearly same antioxidant effects compared to synthetic powerful antioxidant BHT and also expressed powerful antioxidant effect than ${\beta}-carotene$ such as Eucommiae Cortex, Cinnamomi Cortex, Angelicae Gigantis Radix, Lycii Furctus, Acanthopanacis Cortex, Sophorae Radix, Paeoniae Radix, Geranii Herba. Another method of DPPH was performed for searching natural antioxidant from natural product. Sophorae Radix, Puerariae Radix, Aurantii Nobilis Pericarpium, Atractylodis Rhizoma Alba, Acori Graminei Rhizoma, Corni Fructus, Angelicae Gigantis Radix, Paeoniae Radix were carried higher antioxidant capacity than ${\beta}-carotene$ by DPPH method.

• Applied Biological Chemistry
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• 제33권2호
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• pp.116-119
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• 1990

두충껍질의 휘발성 성분을 SDE장치로 추출한 후 GC 및 GC-MS에 의해서 비교분석하였다. 확인된 성분은 49개 성분이며, acid류 4종, alcohol류 11종, aldehyde류 9종, ester류 4종, ketone류 3종, hydrocarbon류 16종, lactone 1종이 확인되었다. 이중에서 가장 많이 함유된 성분은 caproic acid이며, 전체 휘발성 성분의 18.1%를 차지하였다.

• 생약학회지
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• 제6권1호
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• pp.35-38
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• 1975

Attempts were made to investigate the pharmacological actions of Euonymus japonicae Cortex which is used as a substitute of Eucommiae Cortex. Practically the influence of the alcohol extract of the bark on the blood pressure response of rabbit, was examined, and the results are as follows. 1) The intravenous injections of the alcohol extracts of the bark decreased the blood pressure of rabbits to 4.6 11.1 and 34.2 mmHg respectively. 2) The intravenous injection of 10 mg/kg of the alcohol extract after intravenous injection of atropine 4 mg/kg or that of propranolol 2 mg/kg did not give any notable change to the blood pressure. 3) The intravenous injection of 10 mg/kg of the alcohol extract slightly decreased the raised blood pressure caused by the administration of acetylcholine $10^{-8}g/kg\;and\;10^{-7}g/kg$.

• 동의생리병리학회지
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• 제22권6호
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• pp.1454-1461
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• 2008

In oriental medicine, it is known that Eucommiae Cortex (EC) has strengthening and rehabilitative effects on the bone-muscle dysfunction. This study aimed to evaluate the effect of EC on the skeletal muscle atrophy. The muscle atrophy was induced by unilateral transection of the sciatic nerve in Sprague-Dawley rats. EC (water-extract, 170mg/100 g body weight) was treated once a day for 12 days. In this study, the effect of EC examined the muscle weight of hind limb, cross section areas of muscle fibers, fiber type compositions, apoptosis related factors (Bax and Bcl-2). EC reduced muscle atrophy in soleus (SOL), medial gastrocnemius (MGT), extensor digitorum longus, and tibialis posterior significantly in the damaged hind limb. EC increased type-I muscle fibers and decreased type-II muscle fibers significantly in SOL of the damaged hind limb. EC enlarged cross section areas of type-I and type-II muscle fibers significantly in SOL. EC enlarged cross section areas of type-I and type-II muscle fibers significantly in. EC reduced apoptotic nuclei and atrophic muscle fibers in SOL and MGT. EC reduced Bax positive muscle nuclei in SOL and MGT. EC up-regulated Bcl-2 positive muscle fibers in SOL and MGT. These results suggest that EC has an anti-atrophic effect and anti-apoptotic effect against myonuclear apoptosis induced by the peripheral nerve damage.

• Journal of Acupuncture Research
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• 제34권2호
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• pp.1-18
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• 2017

Objectives : The purpose of this study was to evaluate the effects of water extracts of Eucommiae cortex (EC), Psoraleae semen (PS), and their combination on receptor activator of nuclear factor-kappa-B ligand (RANKL)-induced osteoclast differentiation. Methods : We assayed the protein expression levels of nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1), c-Fos, mitogen-activated protein kinases (MAPKs), and ${\beta}-actin$ in cell lysates using western blotting. Similarly, mRNA expression levels of NFATc1, c-Fos, tartrateresistant acid phosphate (TRAP), and glyceraldehyde-3-phosphate dehydrogenase, spermatogeni (GAPDHS) from bone marrow macrophages (BMMs) were analyzed using reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, we determined the anti-osteoporotic effects of the water extracts of EC, PS, and their combination in a lipopolysaccharide (LPS)-induced bone-loss mouse model. Results : The in vitro data revealed showed that the combination of EC and PS extract showed a more remarkable inhibition of osteoclast differentiation than each herb did alone. The combination downregulated the induction of c-Fos, NFATc1, and TRAP by suppressing the phosphorylation of p38 and c-Jun N-terminal kinases (JNKs) and inhibiting nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$). Lastly, the in vivo data showed that PS reduced the LPS-induced bone erosion. Conclusion : The result of this study suggests that EC and PS could be potential therapeutic agents for bone loss diseases such as osteoporosis.

• 대한예방한의학회지
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• 제21권3호
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• pp.87-98
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• 2017

Objectives : The objective of present study is to evaluate anti-arthritic effects of dried pomegranate concentrate powders (PCP), Eucommiae Cortex aqueous (EC) and ethanolic (ECe) extracts, Achyranthis Radix aqueous (AR) and ethanolic (ARe) extracts on the primary cultured rat articular chondrocytes. Methods : MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium Bromide) assay was performed cytotoxic effect of test substances. In addition, anti-inflammatory effects were also observed on the lipopolysaccaride (LPS) treated chondrocytes through prostaglandin $E_2\;(PGE_2)$ production and 5-lipoxygenase (LPO) activities, and inhibitory effects on metalloproteinase (MMP)-2 and MMP-9 activities were observed on the recombinant human interleukin $(rhIL)-1{\alpha}$ treated chondrocytes with their extracellular matrix (ECM) related mRNA expressions - collagen type II, SOX9 and aggrecan. Results : As results, ECe and ARe showed obvious cytotoxicity against primary cultured rat articular chondrocytes at a dose level of 10 mg/ml, respectively. However, no obvious cytotoxic effects of PCP, EC and AR were demonstrated at a dose level of 10 mg/ml, on the primary cultured rat articular chondrocytes. In addition, treatment of LPS $50{\mu}g/ml$ induced significant increases of $PGE_2$ contents and 5-LPO activities indicating inflammatory responses of the primary cultured rat articular chondrocytes, and also decreases of cell viabilities, increases of MMP-2 and MMP-9 activities with decreases of extracellular matrix (ECM) related collagen type II, SOX9 and aggrecan mRNA expressions were observed by treatment of $rhIL-1{\alpha}$ 50 ng/ml, suggesting damages on the primary cultured rat articular chondrocytes and related ECM degradations. However, these inflammatory responses and related ECM degradations were inhibited by pretreatment of all test substances, in order of PCP > ECe > ARe > EC > AR, and $rhIL-1{\alpha}$ induced chondrocytes deaths are inhibited by treatment in order of PCP > EC > AR > ECe > ARe. Conclusions : Taken together, it is expected that mixed formulation of PCP as main components with appropriate proportion of EC and AR as additional components will be achieved a potent alternative medicinal food for osteoarthritis.

• 동의생리병리학회지
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• 제33권2호
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• pp.131-140
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• 2019

The objective of present study is to evaluate concentration-dependent in vitro anti-osteoarthritic (OA) effects of synergic mixed formula consisted of dried pomegranate juice concentrate powder, Eucommiae Cortex aqueous extract and Achyranthis Radix aqueous extract 5:4:1 (g/g) mixture on the primary cultured rat articular chondrocytes. First, any cytotoxic effect of mixture was observed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium Bromide) assay. Next, cyto-protective effect of test substances was evaluated by using the recombinant human interleukin $(rhIL)-1{\alpha}$ induced chondrocytes. In addition, anti-inflammatory effects were also observed on the lipopolysaccaride (LPS) treated chondrocytes through prostaglandin $E_2(PGE_2)$ productions and 5-lipoxygenase (LPO) activities, and inhibitory effects on matrix metalloproteinase (MMP)-2 and MMP-9 activities were observed on $rhIL-1{\alpha}$ treated chondrocytes with their extracellular matrix (ECM) related mRNA expressions. No obvious cytotoxic effects of mixture were demonstrated. Inflammatory damages of chondrocytes and related ECM degradations induced by treatment of LPS or $rhIL-1{\alpha}$ were significantly and concentration-dependently inhibited by pretreatment of mixture from a concentration level of 0.001 mg/ml to 1 mg/ml. In addition, mixture showed $IC_{50}$ for $rhIL-1{\alpha}-induced$ MMP-2 and MMP-9 activities as 44.01 and $162.47{\mu}g/ml$, and also showed $EC_{50}$ for $rhIL-1{\alpha}-induced$ inhibition of collagen type II, SOX9 and aggrecan mRNA expression as 8.61, 10.79 and $4.47{\mu}g/ml$, respectively. It is observed that mixture showed concentration-dependent anti-inflammatory and cytoprotective ECM preserved effects on the primary cultured rat articular chondrocytes without cytotoxicity.