• Journal of Acupuncture Research
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• v.32 no.2
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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.

• Journal of Acupuncture Research
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• v.23 no.3
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• pp.177-190
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• 2006

Objectives : It has long been known about the anticancer effect of GRR-HAS, however, it has not been systemically determined the differentially regulated genes by GRR-HAS in cancer cells. The purpose of this study is to screen the GRR-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cell lines. Oligonucleotide microarray and proteomic approaches were employed to screen the differential expression genes. Methods : GRR~HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. Cells were treated with various concentrations of GRR-HAS (0.1, 0.5, 1.5, 10, $20mg/m{\ell}$) for 24 h. Cell toxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with $1.5mg/m{\ell}$ of GRR-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide genechip (Human genome Ul33 Plus 2.0., Affimatrix Co.). For proteomic analysis, total protein was analyzed by 2D gel electrophoresis and Q-TOF mass spectrometer. Results : It has no cytotoxic effects on both HepG2 cells in all concentrations(0.1, 0.5, 1.5, 10,$20mg/m{\ell}$). In oligonucleotide microarray assay, the number of more than twofold differentially regulated known genes was 320 with 6 up-regulated and 314 down-regulated genes in HepG2 cells. In proteomic analysis, three spots were identified by 2D-gel electrophoresis and Q-TOF analysis. One down -regulated protein was protein disulfide isomerase and up-regulated proteins were fatty acid binding protein 1 and 14-3-3 gan1lTIa protein by $1.5mg/m{\ell}$ of CRR-HAS. Discussion : This study showed the comprehensive gene expression analysis using oligonucleotide microarray for the screening of GRR-HAS mediated differentially regulated genes. These results will provide a better application of GRR-HAS in cancer field and drug target development.

• The Journal of Internal Korean Medicine
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• v.40 no.4
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• pp.697-708
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• 2019

Objective: The aim of this study was to introduce gastric dysmotility as a common cause in patients with concurrent functional dyspepsia and chronic atrophic gastritis. Method: Dyspeptic symptoms, the Rydoraku score, gastric motility (electrogastrography, bowel sound analysis), gastric mucosa (gastroendoscopy), and blood and blood chemistry were all evaluated. For the treatment method, Pyengwi-san (solution) and Banwhasashim-tang (extract) were used as herbal drugs. Both ST36 electrical stimulation and simple immersion stimulation of CV11, 12, and 13 in the abdomen were applied. Results: Dyspeptic symptoms including indigestion, headache, and insomnia were all relieved. Gastric myoelectrical activity and gastric pyloric function were additionally improved. The condition of the gastric mucosa was changed from atrophic to erosive. Other side-effects of the treatment were not noted. Conclusion: The traditional Korean treatment showed effectiveness in the relief of dyspeptic symptoms and mucosal improvement of chronic atrophic gastritis. Gastric dysmotility is a common cause of the condition being concurrent with both functional dyspepsia and chronic atrophic gastritis without Helicobacter pylori infection.

• Journal of Acupuncture Research
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• v.25 no.2
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• pp.1-9
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• 2008

목적 : Acetylcholine은 콜린과 아세트산의 에스테르로 인체에서 중요한 신경전달물질로 Acetylcholine-sterase(AChE)라는 효소에 의해 분해된다. Alzheimer's disease, ataxia, myasthenia gravis, Parkinson' disease 등의 질환에 AChE 억제제가 사용되어 왔으며 최근 한약재의 AChE 억제 효능에 관한 연구들도 진행되고 있다. 봉독은 관절염, 통풍 등의 질환에 응용되어 왔으며 진통효과 및 항염증작용에 대한 임상적, 실험적 연구가 많이 보고되어 왔으나 AChE 억제효과에 대한 연구는 아직까지 보고된 바 없다, 본 연구에서는 봉독약침액과 봉독의 과민반응 유발항원 중 하나인 Phospholipase A2 억제효능이 있는 것으로 알려진 상백피를 혼합한 상백피봉독약침액의 AChE 억제효과를 알아보았다. 방법 : PC12 세포주에서 추출한 AChE와 0.1, 0.01 and $0.001mg/m{\ell}$ 농도의 봉독약침액 및 상백피봉독약침액을 60분간 반응시켰다. 효소면역측정법(ELISA)을 이용하여 흡광도를 10분, 30분, 60분 경과시 각각 측정한 후 효소활성저해도(%)를 계산하였다. 효소활성저해도(%) = [(Cc - Ce)/Cc] ${\times}$ 100 Cc : 대조군 흡광도, Ce : 실험군 흡광도 결과 : 1. 봉독약침액은 0.1, 0.01, $0.001m{\ell}/mg$의 농도에서 30분 경과 후부터 유의성 있는 억제효과를 나타내었다. 2. 상백피봉독약침액은 $0.1m{\ell}/mg$ 농도에서 10분 경과 후부터 유의성 있는 억제효과를 나타내었고, $0.01m{\ell}/mg$ 농도에서 30분경과 시 유의성 있는 억제효과를 나타내었다. 3. 봉독약침액과 상백피봉독약침액의 AChE 억제효과 비교에서 봉독약침액의 억제효과가 상백피봉독약침액 보다 뛰어났다. 요약 : 봉독약침액과 상백피봉독약침액의 AChE 억제효과를 확인하여 두 군 모두 유의성 있는 결과를 얻을 수 있었다. 앞으로 알츠하이머병이나 치매와 같은 신경퇴행성 질환에 대한 봉독의 임상적 활용 및 보다 넓은 범위에 대한 연구가 필요할 것이라 사료된다.

• Journal of Acupuncture Research
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• v.25 no.1
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• pp.139-154
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• 2008

목적 : 홍화(紅花)는 활혈거어(活血祛瘀), 해독지통(解毒止痛)의 효능이 있어 관절염, 동맥경화(動脈硬化), 종양(腫瘍), 월경부조(月經不調), 뇌혈전(腦血栓)에 사용되어 왔다. 이에 홍화약침액(紅花藥鍼液)의 분자생물학적 효능 분석을 하고자 Lipopolysaccharide(LPS)로 염증을 유발한 RAW 264.7 cell의 유전자(遺傳子) 발현(發顯)에 미치는 영향을 Microarray를 통하여 관찰하였다. 방법 : RAW cell을 배양하고 홍화약침액(紅花藥鍼液)의 세포 독성을 확인한 후 (1) LPS, (2) 홍화약침액(紅花藥鍼液), (3) 홍화약침액(紅花藥鍼液)과 LPS를 처치했을 때의 유전자 발현양상을 microarray를 이용하여 관찰하였다. 대조군에 비해 2배 이상 발현의 차이가 있는 경우를 유의한 것으로 보았다. 결과 : 8,170개의 유전자 중 (1) LPS를 처치하였을 경우 35개의 유전자에서 발현이 상승되었고, (2) 홍화약침액(紅花藥鍼液)을 처치하였을 경우 11개의 유전자에서 발현이 상승되고 53개의 유전자에서 발현이 억제되었으며, (3) 홍화약침액(紅花藥鍼液)과 LPS를 동시에 처치하였을 경우에는 47개의 유전자에서 발현이 상승되었고 11개의 유전자에서 발현이 억제되었다. LPS 자극으로 발현이 상승되었지만 홍화약침액(紅花藥鍼液)을 처치할 때 발현이 억제되는 유전자는 SUMO1/sentrin specific protease 7(SENP7), Serine(or cysteine) proteinase inhibitor, clade B(ovalbumin), member 7(SERPINB7), M-phase phosphoprotein, mpp8(HSMPP8), Glycogenin 2(GYG2), InaD-like(Drosophila)(INADL), Copine III(CPNE3), Loss of heterozygosity, 11, chromosomal region 2, gene A(LOH11CR2A), Chromosome 9 open reading frame 33(SHC3), NADH dehydrogenase(ubiquinone) 1 beta subcomplex, 2, 8kDa(NDUFB2)로 9개가 있었다. 요약 : 홍화약침액(紅花藥鍼液)이 LPS로 염증을 유발시킨 RAW 264.7 cell의 유전자 발현에 미치는 영향을 Microarray를 통해 분석하였다. 홍화약침액(紅花藥鍼液)이 LPS로 발현을 항진시킨 35개의 유전자 중 9개를 효과적으로 억제하는 것을 확인하여 염증 치료 기전을 시사하는 유용한 자료를 얻을 수 있었으며 홍화약침액(紅花藥鍼液)이 발현을 항진시킨 유전자들을 통해 혈관생성과 종양억제 등 보다 넓은 범위에 대한 연구가 가능할 것으로 사료된다.

• Journal of Pharmacopuncture
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• v.9 no.2
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• pp.5-16
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• 2006

Objective : It has long been known about the osteogenic effect of CPC-HAS on bone tissues. However, it has not been determined the effect of CPC-HAS on cancer cells. The purpose of this study is to screen the CPC-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cells. Oligonucleotide microarray and proteomics approaches were employed to screen the differential expression genes. Methods : CPC-HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. Cells were treated with various concentrations of CPC-HAS (0.1, 0.5, 1.5, 10, 20mg/ml) for 24 h. Cell toxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with 1.5mg/ml of CPC-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide Genechip(Human genome Ul33 Plus 2.0., Affimatrix Co.). For proteomic analysis, total protein was analyzed by 2D gel electrophoresis and Q-TOF mass spectrometer. Results : It has no cytotoxic effects on both HepG2 cell in all concentrations(0.l, 0.5, 1.5, 10, 20mg/ml). In oligonucleotide microarray assay, the number of more than twofold differentially regulated known genes was 23 with 5 up-regulated and 18 down-regulated genes in HepG2 cells. In proteomic analysis, three spots were identified by 2D-gel electrophoresis and Q-TOF analysis. Two down-regulated proteins were aldehyde dehydrogenase 1 and enolase 1, and up-regulated protein was fatty acid binding protein 1 by 1.5mg/ml of CPC-HAS. Discussion : This study showed the screening of CPC-HAS mediated differentially regulated genes using combined approaches of oligonucleotide microarray and proteomic analysis. The screened genes will be used for the better understanding of the therapeutic effects of CPC-HAS on cancer fields.

• Journal of Acupuncture Research
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• v.19 no.1
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• pp.189-202
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• 2002

Objective : This study is designed to investigate the effects of bee venom and melittin on cell death in neuroblastoma cell line after pretreatment with NG(nerve growth inhibitory substance) Methods : It was evaluated by using MTT assay, morphological method, DNA fragmenation, flow cytometry, immunocytochemistry analysis, RT-PCR and Western blot. Results : The MTT assay demonstrated that neuroblastoma cell viability was significantly inhibited dose-dependently by treatment with bee venom and melittin after pretreatment with NG in comparison awith control. The morphological study and fow cytometry demonstrated that neuroblastoma cell showed apoptosis. DNA fragmenation showed DNA ladder below 1 Kbp. Immunocytochemistry assay demonstrated that Fos and MAPK were down-regulated. RT-PCR analysis demonstrated that Fos and MAPK was down-regulated. Western blot demonstrated that Fos and MAPK were down-regulated from $1{\mu}g/ml$ bee venom in neuroblastoma cell pretreated with NG. Conclusion : These result suggests that bee venom and melittin after NG treatment have significant anti-cancer effect and further study is needed in vivo.

• Journal of Acupuncture Research
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• v.22 no.3
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• pp.227-241
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• 2005

Objective & Methods : The purpose of this study is to observe the effects of Asari Herba Cum Radice herbal-acupuncture solution(AHCR-HAS) on arthritis of mice induced by Collagen II at Joksamni(ST36). The author performed several experimental items. First, it is the cell survival rate of mice lung fibroblasts and expression of TNF-${\alpha}$ in synovial cells. Second, it is the incidence rate of arthritis and the weight of spleen. Third, it is the levels of IL-6, TNF-${\alpha}$, INF-${\gamma}$, IgG, IgM and anti-collagen II in serum Fourth, it is histological analysis of the mice joint. Fifth, it is expression ratio of CD3e+ to CDl9+ cell, CD4+ to CD8+ cell, CD69+/CD3e+ cells, CD11+/CD19+ cells and CD11b+/Gr-1+ cells. Result : 1. The highest survival rate of mice lung fibroblasts were measured in the 1% AHCR-HAS, and the expression of TNF-${\alpha}$ in synovial cells were significantly decreased in the 1% AHCR-HAS. 2. In the AHCR-HA I & AHCR-HAII groups, the incidence of arthritis and the weight of spleen were significantly decreased. 3. In AHCR-HAI & AHCR-HAII groups, the levels of IL-6, INF-${\gamma}$, TNF-${\alpha}$, IgG, IgM and anti-collagen II in serum of CIA mice were significantly decreased. 4. In histology, the cartilage destruction and synovial cell proliferation were decreased in the AHCR-HA I & AHCR-HAII groups, and the collagen fiber expressions in the AHCR-HA I & AHCR-HAII groups were similar with that of the Normal group. 5. In the AHCR-HA I & AHCR-HA II groups, the expression ratio of CD3e+ to CD19+ cell and CD4+ to CD8+ cell were similarly maintained as Normal group in lymph nodes, and CD69+/CD3e+ cells and CD11a+/CD19+ cells were decreased in Iymph nodes, and CD11b+/Gr-1+ cells were decreased in synovium. Conclusion : Taking all these observations into account, AHCR-HA is considered to be effective in prophylaxis and treatment of rheumatoid arthritis, and then more effective in prophylaxis than treatment, so put to practical use in future rheumatoid arthritis clinic.

• Journal of Acupuncture Research
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• v.22 no.3
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• pp.1-12
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• 2005

It was clarified that ethanol extract herb-acupuncture solution (EE-UD) and hydrotherapy herb-acupuncture solution (WE-UD) in Ulmus davidiana Planch (Ulmaceae), are the excellent inhibitors of cathepsin K and L. WE-UD inhibited cathepsin K when IC50 value was 5.32 ${\square}g$/ml, and suppressed cathepsin L when IC50 value was 6.34 ${\square}g$/ml. However, EE-UD indicated the activity of inhibiting cathepsin K and L in the level of 1.45 ${\square}g$/ml and 2.43 ${\square}g$/ml, thus it showed more significance than WE-UD. It could be observed that EE-VD is an excellent inhibitor to cathepsin K with Ki value of 0.8 ${\square}g$/ml. This activity is increased by 10-fold even in the analytical experiment when having operations like glutathione in pH 7.0. Also, this supports the mixture of GSH thiolate anion, thus it was thought that this increase in effectiveness is probably attributable to the enhanced chemical function in the combinations of herb-acupuncture solution towards a place of activity in enzyme. WE-UD showed the time-dependent inhibiting property, thus it allowed to know the disunion and the compounding speed in constant cathepsin K during the process of experiment. Finally, EE-UD was proved to suppress the absorbent bone ash in the experiment related to osteoclast in rats for test, and to the bone in rodent. It was proved that WE-UD has the effect of inhibiting the protease in cathepsin K and L, and in collagen of bone. These results strongly suggest that it is effective in preventing the progress of bone damage, which was induced due to cathepsin K. Also, it obtained the conclusion that it is effective to the reabsorption activity of bone in the bone marrow cells.

• Journal of Pharmacopuncture
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• v.17 no.3
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• pp.25-30
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• 2014

Objectives: This study was performed to evaluate the single-dose intravenous toxicity of Guseonwangdo-go glucose 20% pharmacopuncture. Methods: Forty Sprague-Dawley rats were divided into four groups of five males and five females per group: an intravenous (IV) injection of 1.0 mL of normal saline solution per animal was administered to group 1 (G1, control group); an IV injections of 0.1, 0.5, and 1.0 mL of Guseonwangdo-go glucose pharmacopuncture per animal were administered to experimental groups 2, 3, and 4 (G2, G3, and G4), respectively. General symptoms, body weights, hematological and biochemical test results, and necropsy histopathological observation were recorded in all groups. In the statistical analyses, significance was determined by using the one-way analysis of variance (ANOVA). The significance level was 0.05 in all comparisons. Results: For 14 days, no deaths or abnormalities were observed in any of the 4 groups. The body weights of all groups continuously increased during the observation period. In the hematological test, the WBC count was significantly increased in female rats of G4 compared to the control group, but this difference was considered not to be statistically meaningful. No significant biochemical changes were observed. On necropsy, crust formation was observed in one rat of the control group, and granulation tissues were observed around the injection site in one rat of G4; these changes were concluded to have been caused by injection of the needle into a vein. Conclusion: The findings suggest that the lethal dose of Guseonwangdo-go glucose pharmacopuncture is more than 1.0 mL per animal in both male and female rats. Thus, we can conclude that Guseonwangdo-go glucose pharmacopuncture injection is relatively safe to use in acute toxicity tests. Further studies are needed to establish more detailed evidences of its toxicity.