• Journal of Acupuncture Research
• /
• v.24 no.1
• /
• pp.151-163
• /
• 2007

Objectives : Juglandis Semen herbal acupuncture solution(JSS) has a broad array of clinical applications in oriental medicine, including treatment of chronic musculoskeletal diseases such as arthritis. This study was performed to investigate the global gene expression profiles using microarray assay in RAW 264.7 cell line treated with JSS and to advance our understanding of the pharmacologic effect of JSS. Methods : Change of the gene expression profile in RAW cell line following treatment with lipopolysaccharide(LPS) alone, or with LPS plus JSS was investigated with a cut-off level of 2 fold change in the expression. Especially, Change of the gene expression by treatment with LPS alone was compared with that by treatment with LPS plus JSS with a cut-off level of 1/2 fold change in the expression. Results: Of the 8170 genes profiled in this study, 51 were upragulated and 21 downregulated following LPS treatment, and 88 were upregulated and 69 downregulated following costimulation of JSS and LPS. Of the 51 genes upregulated following LPS treatment, 10 were downregulated following costimulation of JSS and LPS. Of the 21 genes downregulated following LPS treatment, 3 were upregulated following costimulation of JSS and LPS. Conclusion : JSS treatment induced upregulation of some genes including IL-10 and downregulation of that including MMP13 with its possible implication in an antiinflammatory action of JSS. However, further research on expression profile changes induced by JSS treatment is expected.

• Journal of Acupuncture Research
• /
• v.24 no.4
• /
• pp.107-113
• /
• 2007

목적 : 본 연구는 계혈등 약침액이 자궁경부암 세포주 SNU-17에서 세포 사멸 효과가 있는지 알아보고자 하였다. 방법 : 자궁경부암 세포주 SNU-17에서 세포 사멸의 변화를 관찰하기 위해서 MTT cytotoxicity assay, DAPI staining, TUNEL assay, RT-PCR analysis 방법을 이용하였다. 결과 : 세포독성 검사에서 계혈등 약침액은 자궁경부암 세포주 SNU-17에 농도 의존적으로 세포독성을 나타내었다. 이러한 계혈등 약침액의 세포독성이 세포사별로 인한 것인지 다른 기전에 의한 것인지 알아본 결과 계혈등 약침액에 의한 세포독성은 DAPI staining 과 TUNEL assay에서 세포사멸의 특정적인 소견들을 나타내었다. 계혈등 약침액이 Bax, Caspase-3의 발현에 미치는 영향을 RT-PCR로 관찰한 결과 계혈등 약침액은 Bax, Caspase-3의 발현을 증가시켰다. 결론 : 이상의 결과 계혈등 약침액이 자궁경부암 세포주 SNU-17에서 세포 사멸을 야기하여 자궁경부암의 치료에 유용할 것으로 사료된다.

• Journal of Acupuncture Research
• /
• v.19 no.2
• /
• pp.28-38
• /
• 2002

Objective : This study is aimed to investigate the effects of bee venom and purified bee venom on cell death in synovial cell line. Methods : It was evaluated by using MTT assay, morphological method, flow cytometry, immunocytochemistry analysis, RT-PCR. Results : The result obtained is as follows. 1. The MTT assay demonstrated that synovial cell viability was significantly inhibitted dose-dependently by treatment with BV and PBV in comparison with control. And the inhibitory effect of BV and PBV was almost same. 2. The morphologic study demonstrated that synovial cell showed apoptotic body resulted from apoptosis after treatment with BV and PBV for 6 hours using microscope. 3. The Flow cytometry demonstrated that apoptosis of synovial cell treated with BV and PBV was related with stop of cell cycle in stage of G0/G1. 4. Immunocytochemistry assay demonstrated that COX-II and iNOS were slightly expressed by treatment with BV and PBV in comparison with control group. 5. RT-PCR analysis demonstrated that COX-II were almost down-regulated by high dose treatment with BV and PBV in comparison with control group. iNOS were well down-regulated by treatment with $5{\mu}g/ml$ BV and PBV whereas it was well expressed in control group. Conclusion : These results suggest that bee venom and purified bee venom have significant effect on cell death in synovial cell line and further study is needed in vivo.

• Journal of Acupuncture Research
• /
• v.20 no.3
• /
• pp.45-62
• /
• 2003

Objective : To examine the effects of Beevenom on the cell proliferation of human breast carcinoma cell line MCF-7, we performed various experiments such as does-dependent effect of Beevenom on cell proliferation and viability, morphological changes, and alterations of apoptosis/cell cycle-regulatory gene products. Methods : Beevenom induced cell viability and proliferation of MCF-7 cells in a concentration-dependent manner. The anti-proliferative effect by Beevenom treatment in MCF-7 cells was associated with morphological changes such as membrance shrinking and cell rounding up. Results : Beevenom induced apoptotic cell death in a concentration-dependent manager, which was associated with degradation of ${\beta}$-catenin, an apoptotic target protein. Beevenom induced the Bax expressions, a pro-apoptotic gene, both in protein and mRNA levels, however, the levels of Bcl-$X_{S/L}$ expression, an anti-apoptotic gene, were down-regulated in Beevenom-treated cells. Western blot analysis and RT-PCT data revealed that the levels of cyclin of B1 protein and cyclin E mRNA were reduced by Beevenom treatment in MCF-7 cells, respectively, where as the expression of tumor suppressor p53 and cyclin dependent kinase inhibitor p21 mRNA were markedly increased in a concentration-dependent fashion. Conclusions : Taken together, these findings suggest that Beevenom induced inhibition of human breast cancer cell proliferation is associated with the induction of apoptotic cell death and Beevenom may have therapeutic potential in human breast cancer.

• Journal of Acupuncture Research
• /
• v.26 no.3
• /
• pp.27-38
• /
• 2009

목적 : 이 연구는 Vipera lebetinat turanica의 사독약침액(蛇毒藥鍼液)(Snake venom toxin, SVT)이 인간 유방암 세포주인 MCF-7과 MDA-MB-231 세포에서 암세포성장의 억제 및 그 기전에 대하여 살펴보고자 하였다. 방법 : SVT를 처리한 후 MCF-7과 MDA-MB-231의 성장억제를 관찰하기 위해 CCK-8 assay를 시행하였고, apoptosis 평가에는 TUNEL assay를 시행하였다. 세포자멸사 관련 세포기전을 보기 위하여 세포내 활성산소량 및 미토콘드리아의 세포막전위 변화를 측정하였고, 세포자멸사 조절 단백인 Bax, Bcl-2 발현 변화 관찰에는 westem blot analysis를 시행하였다. 결과 : MCF-7과 MDA-MB-231 세포에 SVT를 처리한 후, 유방암 세포의 성장, Apoptosis의 유발 및 기전에 미치는 영향을 관찰하여 다음과 같은 결과를 얻었다. 1. MCF-7 세포와 MDA-MB-231 세포에서 SVT를 처리한 후 유방암 세포 성장이 억제되었다. 2. TUNEL assay를 통한 세포자멸사 평가에서 SVT를 처리한 MCF-7세포와 MDA-MB-231 세포 모두 세포자멸사 활성세포의 유의한 증가를 나타내었다. 3. 세포자멸사 관련 세포기전연구에서 SVT를 처리한 MCF-7 세포와 MDA-MB-231 세포에서 세포내 활성산소의 유의한 증가와 미토콘드리아 세포막 전위의 유의한 변동이 관찰되었다. 4. SVT를 처리한 MCF-7세포와 MDA-MB-231세포는 세포자멸사 관련 단백 발현에서 Bax의 유의한 증가와 Bcl-2의 유의한 감소를 나타내었다. 결론 : 이상의 결과는 SVT가 세포내 활성산소를 증가시킴으로써 미토콘드리아의 세포막전위에 변화를 일으켜 유방암 세포주인 MCF-7과 MDA-MB-231 세포에 세포자멸사를 유발하여 증식억제 효과가 있음을 입증한 것이다.

• The Journal of Internal Korean Medicine
• /
• v.20 no.1
• /
• pp.111-121
• /
• 1999

Cervus elaphus (CE), being known to reinforce Kidney, have tested to study the effects concerning damages of renal tissue induced by oxygen free radicals. I had observed the effects of CE extract on damages of rat's kidney following ischemia and reflow. Before ischemia was caused, CE extract was applied $0.2m{\ell}$ per 250g through femoral vein in ischemia and reflow group and normal sailine was applied in normal group, Ischemia was caused by renal artery's clamp for 60 min and reflowed by clamp remove after 30 min. It was increased on the content of lipid peroxidation, activies and type conversion ratio of xanthine oxidase following ischemia and reflow. by clamp remove after 30 min. It was increased on the content of lipid peroxidation, activies and type conversion ratio of xanthine oxidase following ischemia and reflow. However, they were decreased when CE extract was pre-applied. Glutathione level was decreased in ischemia and reflow group, and increased in CE extract's pre-applied group. However, it could not seen special changes on aldehyde oxidase activities, either. In conclusion, CE extract recovers the damage of kidney due to ischemia and reflow by decreasing the lipid peroxidation.

• Korean Journal of Acupuncture
• /
• v.20 no.1
• /
• pp.45-55
• /
• 2003

Backgrounds : It has been investigated about ageing theory. However, ageing mechanism still remains to be unknown. Since Harman proposes that ageing and ageing related diseases might be due to oxidative damage and these are modifiable by genetic and environmental factors. For designing an optimal nutritional countermeasure against ageing and ageing related disease, it is necessary to understand the ageing mechanism and other reactive species. Objectives : Schizandra Chinesis has been clinically used to treat brain disease, respiratory or inflammatory disease etc. in Oriental medicine. The purpose of this study is to investigate the scavenging effect of Schizandrae Fructus herbal-acupuncture solution(SFHA) on NO, DPPH and IL-4. Results : The results are summarized as follows; (1) There is a significant scavenging effect of SFHA on NO in 0.1, 1, 10mg/ml group after 24hrs. (2) There is a significant scavenging effect of SFHA on DPPH in 1, $10{\mu}g/ml$ group. (3) There is a significant inhibitory effect of SFHA on IL-4 in 1, 10, $100{\mu}g/ml$ group. Conclusions : These findings indicate that SFHA can be used as antioxidant or antiimflammatory drug. But further study is needed about the effect of SFHA.

• Korean Journal of Acupuncture
• /
• v.18 no.1
• /
• pp.1-9
• /
• 2001

Artemisia asiatica Nakai aqua-acupuncture solution (ANAS) was administered once daily for 10 days before the tumor implantation ($1{\times}10^6\;cells$). Body weight, spleen weight and the number of ascitic tumor cells were measured at 6 days after tumor implantation. The change of body weight and the survival rate of mice were observed for 21 days. It was used three biomarkers (quinone reductase, glutathione, glutathione S-transferase) to test chemopreventive potentials of ANAS. ANAS exerted antitumor activity by inhibiting the growth of Ehrlich ascites tumor cells in vivo. Mice given Ehrlich cells and ANAS at $CV_{12}$ and $BL_{18}$ had 57.1% to 49.2% survival after 21 days. Quinone reductase activity and glutathione levels were increased with ANAS. However, glutathione S-transferase level was 1.1-fold with ANAS. These results suggest that ANAS has chemopreventive potential by inducing QR activity and increasing GSH level.

• Korean Journal of Acupuncture
• /
• v.23 no.2
• /
• pp.125-136
• /
• 2006

Objectives: It has long been known about the osteogenic effect of CPC-HAS(cervi parvum cornu herbal-acupuncture solution) 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 SNU484 gastric cancer cell lines. Oligonucleotide microarray approache was 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, 20 mg/ml) for 24 h. Cell toxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with 1.5 mg/ml of CPC-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide Genechip(Human genome U133 Plus 2.0., Affimatrix Co.). Results: It has no cytotoxic effects on SNU484 cell in all concentrations(0.l, 0.5, 1.5, 10, 20 mg/ml). In oligonucleotide microarray assay, in SNU484 cells, the number of more than twofold up-regulated genes was 5 while, the number of more than twofold down-regulated genes was 10. Conclusions: This study showed the screening of CPC-HAS mediated differentially regulated genes using combined approaches of oligonucleotide microarray. The screened genes will be used for the better understanding of the therapeutic effects of CPC-HAS on cancer fields.

• Korean Journal of Acupuncture
• /
• v.22 no.2
• /
• pp.127-149
• /
• 2005

This study is designed to investigate the method of manufacturing herbal acupuncture through literature of oriental medicine. The findings of this study are as follows; 1. The methods of manufacturing herbal acupuncture go through the process of abstraction, purification, mixing, filtration, putting and tight sealing in the container, sterilization, quality control, printing and packing 2. There are many ways to manufacturing herbal acupuncture, for example water-alcohol precipitation, alcohol-water precipitation, liquid-liquid abstract, acid-base abstract, metal base precipitation, distillation, molecular structure, polyamide absorption, dialysis, and ion exchange, etc. And popular method is water-alcohol precipitation. This is through alcohol precipitate extracting the principal ingredients from water abstraction. This is very simple and efficient way using melting characteristics of compounds in herb to water and ethanol. 3. Sterilization of herbal acupuncture is through heating-pressure, boiling, steam flowing, low temperature, filtering, radiation, cooling, and microwaves. Nowadays filtering is commonly used. And sterilization is estimated by an examination of asepsis . 4. Herbal acupuncture must be undergo study and experiment to clinical use. The problems of herbal acupuncture are turbidity, instability, causing hemolysis, pain, and fever. So many provisions (addition, sterilization, and filtration etc.) must be prepared. 5. The theory of manufacturing herbal acupuncture is from oriental medicine, not western. So it must be corresponded to oriental medical theory, for example Gimi(氣味), Guigyung(歸經), Ingyung(引經), Bosa(補瀉), and Match of Herb. It is recommended that further study of many other sided investigations in the future.