• Journal of Acupuncture Research
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• 제23권4호
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• pp.205-218
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• 2006

Objectives : This study was performed to investigate the effects of anti-cancer and changes In immune response of Lonicerge Flos Herbal-acupuncture. Methods Experimental studies were evaluated through the anti-cancer and immune response activities such as, cell viability, BNA fragmentation, Apoptosis, survival time, pulmonary colonization, and productivity of interleukins & $interferon-{\gamma}$. In order to study the effects of anti-cancer and changes in immune response of Lonicerae Flos Herbal-acupuncture, the groups were divided into five groups ; Normal group(non treated group), Control A group(0.2ml Normal saline for oral administration), Control B group(administration of intramuscular injection with 0.2ml Lonicerae Flos Herbal-acupuncture solution), Acupuncture group(AT, administration of acupuncture at Chungbu(L1)), and Herbal-Acupuncture group(HAT, administration of Lonicerae Flos Herbal-acupuncture at Chungbu(L1)). Results : 1. Lonicerae Flos Herbal-acupuncture(>300mg/ml) could lead cancer cell to cell death. 2. Lonicerae Flos Herbal - acupuncture (40mg/ml) caused DNA cleavage. 3. Lonicerae Flos Herbal-acupuncture(400mg/ml) caused apoptosis in the cancer cell line. 4. In mouse survival time, all of experimental groups didn't show any significant compared to the control group. 5. In pulmonary colonization assay, Lonicerae Flos Herbal-acupuncture group was less than Control A group at 7 days after induction of cancer. 6. In comparison Control A group, there was significant decrease of Interleukin-2 level in Lonicerae Flos Herbal-acupuncture group. 7. In comparison Control group, there was decrease of Interleukin-4 level in the Acupuncture group. 8. In comparison Control group, there was decrease of Interleukin-10 level in the Acupuncture group. 9. In comparison Control group, there was significant increase of Interleukin-12 level in Acupuncture group and Lonicerae Flos Herbal-acupuncture group. 10. In comparison Control group, there was significant increase of $Interferon-{\gamma}$ level in Acupuncture group. Conclusion : According to above mentioned results, Lonicerae Flos Herbal- acupuncture is expected to be effective for anticancer and improvement in immune response.

• 대한본초학회지
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• 제22권2호
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• pp.35-43
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• 2007

Objectives : The purpose of this study was to investigate the effect of Bo-du-san (BOS) on apoptosis in human gastric cancer cells, SNU-l cells. BOS, a drug preparation consisting of two herbs, that is, Crotonis Fructus (Strychni ignatii Semen, bodu in Korean) and Glycyrrhizae Radix (Glycyrrhizae uralensis FISCH, Gamcho in Korean). Methodss : In this study, methanol extract of BOS was examined for cytotoxic activity on human gastric cancer cells, SNU-1 cells, using XTT assay, with an IC50 value was 0.7 mg/ml and 0.3 mg/ml at 24 hrs and 48 hrs, respectively. Apoptosis induction by BDS in SNU-l cells was verified by the induction of DNA fragmentation, cleavage of poly ADP-ribose polymerase (PARP), and activation of caspase-3, -8 and -9. Inhibitors of caspase-3, -8 and -9 (Ac-DEVD-CHO, Z-IETD-FMK and Z-LEHD-FMK) efficiently blocked BOS-induced cell death of SNU-l. Resultss : BOS-induced cell death was via caspase dependent apoptosis. Moreover, treatment of BOS result in the decrease the G1/S cycle regulation proteins (cyclin D1 and E) expression and increase CDK inhibitor proteins (p21 and p27) expression, and increase apoptotic protein, p53 expression. Thus, BOS induces apoptosis in SNU-1 cells via cell cycle arrested in G1 phase. Conclusions : These results indicated that BOS has some potential for use as an anti-cancer agent.

• BMB Reports
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• 제40권4호
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• pp.571-576
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• 2007

Three anti-apoptosis genes, Ls-iap2, iap3 and p49 were found in Leucania separata multiple nuclear polyhedrovirus. Amino acid sequence homology of Ls-IAP2 and Ls-IAP3 with Op-IAP2 and Op-IAP3 from Orgyia pseddotsugata MNPV were 20% and 42%, while that of Ls-P49 is 28% with Sl-P49 from Spodoptera littorolis MNPV. Ls-IAP2 contains one baculoviral IAP repeat (BIR) domain followed by a RING domain, while Ls-IAP3 contains two BIRs and a RING. Ls-P49 contains a reactive site loop, predicted cleavage site (KKLD$^{74}{\downarrow}$G) that is different from Sl-P49 (TVID$^{94}{\downarrow}$G). Expressed Ls-iap3 or Ls-p49 under presence of actinomycin D in SF9 cells, DNA ladder assayrevealed that Ls- IAP3 or Ls-P49 could block the apoptosis of SF9 cells induced by actinomycin D. Replication of p35 deficient-mutant Autographa californica MNPV in SF9 cells was also rescued when Ls-iap3 or Ls-p49 was expressed transiently. No anti-apoptotic activity was observed for Ls-IAP2. The results showed that both of Ls-IAP3 and Ls-P49 were functional apoptotic suppressors in SF9 cells.

• 동의생리병리학회지
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• 제18권6호
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• pp.1617-1621
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• 2004

The aim of this study was to investigate the apoptotic effect and its mechanism on Radix Paeoniae Alba Extract(RPAE) in DU145 human prostate cancer cell line. RPAE induced apoptosis in a dose-dependent manner in DU145 cells as confirmed by both discontinuous DNA fragmentation using Hoechst33342 staining and poly-(ADP-ribose) polymerase(PARP) cleavage, which are apoptotic signs. To clarify the mechanisms on RPAE-induced apoptosis, we examined the p50(NF-κB subunit), IκBα, PTEN and Par-4 protein expression using Western blotting. Treatment with RPAE resulted in the decrease of p50 expression by IκBα increase, which resulted in Par-4 increase and bcl-2 decrease in DU145 cells. These results suggest that apoptosis of DU145 cells by RPAE involved decreases of NF-κB activation and bcl-2 expression, increase of Par-4 protein expression.

• Biomolecules & Therapeutics
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• 제17권3호
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• pp.282-287
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• 2009

Ochnaflavone is a natural biflavonoid and mainly found in the caulis of Lonicera japonica (Caprifoliaceae). Biological activities such as anti-inflammatory and anti-atherogenic effects have been previously reported. The anticancer activity of ochnaflavone, however, has been poorly elucidated yet. In the present study, we investigated the effect of ochnaflavone on the growth inhibitory activity in cultured human colon cancer cell line HCT-15. Ochnaflavone inhibited the proliferation of the cancer cells with an $IC_{50}$ value of $4.1{\mu}M$. Flow cytometric analysis showed that ochnaflavone arrested cell cycle progression in the G2/M phase, and induced the increase of sub-G1 peak in a concentration-dependent manner. Induction of cell cycle arrest was correlated with the modulation of the expression of cell cycle regulating proteins including cdc2 (Tyr15), cyclin A, cyclin B1 and cyclin E. The increase of sub-G1 peak by the higher concentrations of ochnaflavone (over $20{\mu}M$) was closely related to the induction of apoptosis, which was evidenced by the induction of DNA fragmentation, activation of caspase-3, -8 and -9, and cleavage of poly-(ADP-ribose) polymerase. These findings suggest that the cell cycle arrest and induction of apoptosis might be one possible mechanism of actions for the anti-proliferative activity of ochnaflavone in human colon cancer cells.

• 한국수정란이식학회지
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• 제17권2호
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• pp.109-115
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• 2002

The present study was conducted for the production of transgenic cloned cows those secrete human lactoferricin into milk by somatic cell nuclear transfer (NT). To estimate detrimental effects of gene transfection on transgenic cloned embryo production, development rates of NT embryos were compared between transfected and non-transfected cumulus and ear fibroblast cells. An expression plasmid for human lactofericin (pbeta-LFC) was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and human lactoferricin target gene into a pcDNA3 plasmid. Two bovine somatic cell lines (cumulus cell and ear fibroblast) were established and transfected with the expression plasmid using a liposomal transfection reagent, Fugene6 as a carrier. Cumulus cell and ear fibroblast were transfected at the passage of 2 to 4, trypsinized and GFP-expressing cells were randomly selected and used for somatic cell NT. Developmental competences (rates of fusion, cleavage, and blastocyst formation) in bovine transgenic somatic cell NT embryos reconstructed with non-transfectecd cells were significantly higher than those from transfected cells in cumulus cell and ear fibroblast (P＜0.05). This study indicated that transfection of done. cell has detrimental effect on embryo development in bovine transgenic NT.

• 한국수정란이식학회지
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• 제17권2호
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• pp.101-108
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• 2002

Human Prourokinase (proUK) offers potential as a novel agent with improved fibrin specificity and, as such, may offer advantages as an attractive alternative to urokinase that is associated with clinical benefits in patients with acute peripheral arterial occlusion. For production of transgenic cow as human proUK bioreacotor, we conducted this study to establish efficient production system for bovine transgenic embryos by somatic cell nuclear transfer (NT) using human prourokinase gene transfected donor cell. An expression plasmid for human prourokinase was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and human prourokinase target gene into a pcDNA3 plasmid. Cumulus cells were used as donor cell and transfected with the expression plasmid using the Fugene 6 as a carrier. To increase the efficiency for the production of transgenic NT, development rates were compared between non-transfected and transfected cell in experiment 1, and in experiment 2, development rates were compared according to level of GFP expression in donor cells. In experiment 1, development rates of non-transgenic NT embryos were significantly higher than transgenic NT embryos (43.3 vs. 28.4％). In experiment 2, there were no significant differences in fusion rates (85.4 vs. 78.9％) and cleavage rates (78.7 vs. 84.4％) between low and high expressed cells. However, development rates to blastocyst were higher in low expressed cells (17.0 vs. 33.3％), and GFP expression rates in blastocyst were higher in high expressed cells (75.0 vs. 43.3％), significantly.

• 대한한방내과학회지
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• 제31권4호
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• pp.693-705
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• 2010

Objectives : The water extract of Daechilgi-tang(DCGT) has traditionally been used for treatment of qi stagnation(氣滯), which is considered to be one of the important causes of neuronal disease in oriental medicine. However, little is known about the mechanism by which DCGT protects neuronal cells from brain cell damages. Methods and Results : The author tested the mechanism of the cytoprotective effect of DCGT on glutamate -stimulated rat C6 glial cells. DCGT significantly protected C6 glial cells from glutamate in MTT assay. Pre-treatment of C6 glial cells with DCGT markedly inhibited the DNA fragmentation of C6 cells induced by glutamate. Glutamate increased the generation of reactive oxygen species(ROS) and intracellular calcium level in C6 glial cells. However, pre-treatment with DCGT markedly suppressed the increase of ROS generation and intracellular calcium accumulation induced by glutamate. Among apoptosis signaling mediators, DCGT markedly increased the expression level of Bcl2 in glutamate-treated cells. It also inhibited the cleavage of caspase-3 and PARP proteins by glutamate in C6 glial cells. Conclusions : These results suggest that DCGT protects brain cells from glutamate cytotoxicity through inhibition of ROS generation and activation of apoptosis signaling pathway as well as induction of the anti-oxidant system.

• Biomolecules & Therapeutics
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• 제18권4호
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• pp.454-462
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• 2010

The present study investigated the neuroprotective effect of Carpinus tschonoskii MAX and its intracellular protective mechanism on 6-hydroxydopamine (6-OHDA)-induced oxidative damage in PC12 cells. We found that pretreatment of PC12 cells with C. tschonoskii extract significantly inhibited the cell death induced by 6-OHDA in a dose dependent manner. C. tschonoskii extract decreased 6-OHDA-induced apoptotic events such as chromatin condensation, DNA fragmentation, the decrease of Bcl-2/Bax ratio, caspase-3 activation and PARP cleavage. C. tschonoskii extract also reduced generation of 6-OHDA-induced reactive oxygen species and nitric oxide. Furthermore, C. tschonoskii extract up-regulated the myocyte enhancer factor 2 D (MEF2D), a critical transcription factor for neuronal survival, and Akt activity, whereas it inhibited the activity of ERK1/2 and JNK. The results suggest that C. tschonoskii extract decreases 6-OHDA-induced oxidative stress and could prevent PC12 cell apoptosis induced by 6-OHDA via the up-regulation of MEF2D and Akt activity, and thus may have application in developing therapeutic agents for Parkinson's disease.

• 생약학회지
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• 제32권4호통권127호
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• pp.274-279
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• 2001

Ailanthus altissima belonging to the family Simaroubaceae has been used to settle an upset stomach, to combat a fever, and as an insecticide. Apoptosis is an active process, which is a critical feature of the regulated development of multicellular organisms. We investigated whether the extract of A. altissima induced apoptotic cell death in Jurkat T-acute lymphoblastic leukemia (ALL) cells. Upon treatments with the extract, the dose-dependent inhibitions of cell viability were observed. It also caused apoptosis as measured by cell morphology and DNA fragmentation. The capability of the extract to induce apoptosis was associated with proteolytic cleavage of specific target protein such as poly(ADP-ribose)polymerase (PARP) protein, suggesting the possible involvement of the activations of caspases. Further study showed that Bcl-2 protein levels were not changed in all treated groups compared to control group. These results suggest that A. altissima induces Bcl- 2-independent apoptosis in Jurkat T cells.