• 한국가시화정보학회지
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• 제5권1호
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• pp.9-11
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• 2007

유(流)세포분석기(flow cytometer)는 일정한 체적 내에 존재하는 세포의 종류 및 개체 수 등을 계측하는 장비로써 생체에서 추출한 유액상태(혈액 또는 림프액)의 세포를 모세관(micro-channel)을 통과시킬 때 발생하는 산란 및 형광 빛을 이용하여 계측한다. 유세포 분석기는 신약의 투석 후 세포수의 증감, 암세포의 전이 및 세포주기의 분석 등을 연구하는 데 사용되며 현재 Becton-Dickinson's 등에서 상용화된 제품을 생산 판매하고 있으며, 계측을 위해서는 생체에서 세포를 추출해야 한다는 단점을 가지고 있다. Harvard 의과대학에서 최근에 개발한 생체 유세포분석기(In vivo Flow Cytometer)는 생체에서 세포를 추출하지 않고 세포의 수를 계측할 수 있다[1]. 레이저가 혈관의 특정한 부위에 조사되고 있고, 이곳을 세포가 통과하면서 발생하는 형광을 계측함으로써 주어진 시간 동안 특정세포군이 얼마나 지나가는 지를 계측할 수 있는 장비이다. 본 특별기사에서는 혈류 가시화 분야의 독자를 위해 최근에 "Optics Express"에 "In vivo imaging flow cytometer"라는 제목으로 최근에 개제된 논문의 내용을 하여 소개한다[2].

• 생명과학회지
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• 제19권5호
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• pp.620-624
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• 2009

동면 개시인자로 알려진 DADLE는 여러 연구에 의해 in vivo와 in vitro 상에서 유사 동면 상태를 야기한다. 본 연구는 인체혈구암세포인 U937 세포주의 세포 사멸과 세포 주기 둥에 대한 DADLE의 영향을 살펴보았다. DADLE가 처리된 U937세포는 8${\sim}6$10 ${\mu}$M의 높은 농도에서 세포 증식이 감소하였으며, 0${\sim}6$ ${\mu}$M의 낮은 농도에서 영향이 없었다. DNA flow cytometer를 이용하여 세포 주기를 분석해본 결과 DADLE에 의한 세포 주기 억제가 관찰되었다. DADLE처리에 따른 세포 증식률 감소 및 세포 주기 억제효과를 전사 수준에서 조사한 결과 Bcl-XL, c-IAP-2의 발현 및 survivin의 발현 감소가 관찰되었으며, COX-2의 발현 역시 COX-1의 변화 없이 감소함을 확인하였다. 또한, cyclin E 와 cdk-2, -4 그리고 -6의 발현 역시 감소하는 것을 관찰하였다. Telomere 조절 관련 유전자의 경우도 c-myc과 TERT의 감소, 그리고 TEP-1가 증가하는 현상을 관찰하였다. 이상의 결과는 DADLE를 U937 암세포주에 처리했을 때 세포 주기의 억제를 통하여 life-time을 증가시킬 가능성을 시사하며 이에 관한 지속적인 연구가 필요할 것으로 사료된다.

• 생약학회지
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• 제28권4호
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• pp.286-292
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• 1997

We studied the screening condition for immune regulatory responsor. We focused on the T-lymphocytes leer this purpose. Mouse fetal thymic organ culture (FTOC) system and flow cytometric analysis were mainly used in this experiment. Even if FTOC is carried out in vivo condition, the pattern of thymic development in the condition of FTOC is similar to that of in vivo condition. In this regard, FTOC system might be very powerful tool to screen the immune regulator, especially concerning on T cells. To establish the optimum condition of FTOC to screen the Immune regulator, we focused on the optimum amount of dose and culture period. The cell number and surface antigens on T cells were also analysed by using hemacytometer and flow cytometer. To monitor the differentiation event, anti-CD3, anti-CD4 and anti-CD8 antibodies were used. Alkoxyglycerol and Phellodendri Cortex were used fur positive and negative control, respectively. Astragalus membranceus was used as test sample. From our analysis, we reached to conclusions that the best dose of extract is $50\;{\mu}g/ml$ of culture medium, the best culture period is for 9 days, and ethanol used as solvent has no toxicity to FTOC.

• 대한한방내과학회지
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• 제26권1호
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• pp.143-155
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• 2005

Objective : The purpose of this research is to examine the effects of Gleditsiae Spina (GS) extract on immune cells and cytokines in ovalbumin (OVA)-induced asthmatic mice. Methods : In vivo, C57BL/6 mice were sensitized and handicapped by OVA for 12 weeks. During this experiment, the one group was then treated with GS extract for the later 8 weeks (3 times per week) and analyzed by ELISA, flow cytometer and RT-PCR. Results : In vivo, there were significant decreases in eosinophils, IL-4, IL-5, IL-13, IgE in BALF (bronchoalveolar lavage fluid). However, $IFN-{\gamma}$ in BALF of GS group increased significantly, compared with that of control group. Additionally, the population of $CD3e^-/CCR3^+,\;CD69^+/CD3e^+,\;IgE^+/B220^+,\;CD11b^+/Gr-l^+$ cells in the GS group decreased. Conclusion : The results of this study support a role for GS as an effective treatment for asthma in its experimental success in significantly decreasing inflammation and asthma reactions, and in increasing $IFN-{\gamma}$, which helps prevent such reactions.

• 대한한방내과학회지
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• 제26권1호
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• pp.156-168
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• 2005

Objective : The purpose of this research is to examine the effects of Pinelliae Rhizoma (PR) extract on immune cells and cytokines in ovalbumin (OVA)-induced asthmatic mice. Methods : In vivo, C57BL/6 mice were sensitized and handicapped by OVA for 12 weeks. During this experiment, the one group was then treated with PR extract for the 8 weeks (3 times per week) and analyzed by ELISA and flow cytometer. Results : In vivo, there were significant decreases in eosinophils, IL-4, IL-5, IL-13, IgE in BALF (bronchoalveolar lavage fluid) compared with that of control group. However, $IFN-{\gamma}$ in BALF of GS group increased significantly, compared with that of control group. Additionally, the population of $CD3e^-/CCR3^+,\;CD69^+/CD3e^+,\;IgE^+/B220^+,\;CD11b^+/Gr-l^+$ cells in the PR group decreased, compared with that of control group. Conclusion : The results of this study support a role for PR as an effective treatment for asthma in its experimental success in significantly decreasing inflammation and asthma reactions, and in increasing $IFN-{\gamma}$, which helps prevent such reactions.

• 동의생리병리학회지
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• 제19권2호
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• pp.446-451
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• 2005

This experiment was designed to investigate the effect of Mominis Placenta herbal acupuncture solution(HP-HAS) on immune cells and cytokines in murine asthma model. In vivo C57BL/6 mice were sensitized and challenged with OVA for 12 weeks. The experimental group was treated with Hominis Placenta herbal acupuncture solution(HP-HAS) at P'yesu(BL13) for the later 8 weeks(3 times a week) and analyzed by ELISA, flow cytometer. The results were obtained as follows Eosinophils in BALF(bronchoalveolar lavage fluid) of HP-HAS group decreased significantly compared with that of control group. IL-4, IL-5, IL-13, IgE in BALF of HP-HAS group decreased significantly compared with that of control group. Number of $CD3e^-/CCR3^+$, $CD69^+/CD3e^+$, $CD11b^+/Gr-1^+$ cells in the HP-HAS group decreased compared with that of control group.

• 대한한방내과학회지
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• 제27권1호
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• pp.114-125
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• 2006

objective : The purpose of this research is to examine the effects of Scutellaria Radix(SR) extract on immune cells and cytokines in ovalbumin(OVA)-induced asthmatic mice. Methods : In vivo, C57BL/6 mice were sensitized and handicapped by OVA for 9 weeks. In this experiment, one group was not treated, and the other group was treated with SR extract for six weeks(five times per week) and analyzed by ELISA and flow cytometer. Results : In vivo, there were significant decreases in eosinophils, IL-4, IL-13 in BALF (bronchoalveolar lavage fluid) compared with that of control group. However,$IFN{\gamma}$ in the SR group increased significantly compared with that of control group. Additionally, the population of $CD3e^-/CCR3^+,\;CD69^+/CD3e^+,\;IgE^+/B220^+,\;CD11b^+/Gr-1^+$ cells in the SR group decreased compared with that of control group. conclusion : The results of this study support a role for SR as an effective treatment for asthma in its experimental success in significantly decreasing inflammation and asthma reactions, and in increasing $IFN{\gamma}$, Which helps prevent such reactions.

• Journal of Microbiology and Biotechnology
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• 제20권1호
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• pp.224-228
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• 2010

The encapsulation of bacteria may be used to harness them for longer periods of time in order to make them viable, whereas antibiotic treatment would result in controlled release of therapeutic molecules. Encapsulated Escherichia coli GFP (green fluorescent protein) (E. coli GFP) was used here as a model for therapeutic substance - GFP fragments release (model of bioactive substances). Our aim was to evaluate the performance of bacteria encapsulated in hollow fibers (HFs) treated with antibiotic for induction of cell death. The polypropylene-surface-modified HFs were applied for E. coli encapsulation. The encapsulated bacteria were treated with tetracycline in vitro or in vivo during subcutaneous implantation into mice. The HF content was evaluated in a flow cytometer, to assess the bacteria cell membrane permeability changes induced by tetracycline treatment. It was observed that the applied membranes prevented release of bacteria through the HF wall. The E. coli GFP culture encapsulated in HF in vitro proved the tetracycline impact on bacteria viability and allows the recognition of the sequence of events within the process of bacteria death. Treatment of the SCID mice with tetracycline for 8 h proved the tetracycline impact on bacteria viability in vivo, raising the necrotic bacteria-releasing GFP fragments. It was concluded that the bacteria may be safely enclosed within the HF at the site of implantation, and when the animal is treated with antibiotic, bacteria may act as a local source of fragments of proteins expressed in the bacteria, a hypothetical bioactive factor for the host eukaryotic organism.

• 동의생리병리학회지
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• 제17권2호
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• pp.403-411
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• 2003

For evaluating the effect of Siegesbeckiae Herba (here after abbreviated as SBH) on rheumatoid arthritis, the experiment was carried out; Incidence of rheumatoid arthritis, IRA(indices of rheumatoid arthritis), immunophenotypes by flow cytometer and histopathological changes in MRL/MpJ-Ipr-Ipr Mice in vivo were studied. The results were obtained as follows: 1. Incidence of RA in MRL/Ipr mice was suppressed to 60% of control by SBH. 2. IRA was significantly reduced for IgG3 and IgM at 20 weeks of age and for IgG2b at 12 and 20 weeks of age in mice by SBH compared with control. 3. Immunophenotypes such as CD4/sup +//CD25/sup +/, CD8/sup +//CD3e/sup +/, CD69/sup +//B220/sup +/, NK/sup +//CD3e/sup +/ were significantly increased by SBH compared with control. 4. In histopathological analysis, SBH suppressed the progression of PMN(polymorphonuciear leukocyte), leukocyte and fibroblast infiltration, and subsynovial soft tissue edema frequently showing in the early stage of the arthritis, and also effectively reduced the degeneration of cartilages and degenerative bone symptoms. These results suggest Siegesbeckiae Herba can be effectively applied to the treatment of rheumatoid arthritis and it is still necessary to isolate effective compound from Siegesbeckiae Herba in the near future.

• Journal of Ginseng Research
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• 제43권4호
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• pp.635-644
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• 2019

Background: To increase the pharmacological effects of red ginseng (RG, the steamed root of Panax ginseng Meyer), RG products modified by heat process or fermentation have been developed. However, the antiallergic effects of RG and modified/fermented RG have not been simultaneously examined. Therefore, we examined the allergic rhinitis (AR)-inhibitory effects of water-extracted RG (wRG), 50% ethanol-extracted RG (eRG), and bifidobacteria-fermented eRG (fRG) in vivo. Methods: RBL-2H3 cells were stimulated with phorbol 12-myristate-13-acetate/A23187. Mice with AR were prepared by treatment with ovalbumin. Allergic markers IgE, tumor necrosis factor-${\alpha}$, interleukin (IL)-4, and IL-5 were assayed in the blood, bronchoalveolar lavage fluid, nasal mucosa, and colon using enzyme-linked immunosorbent assay. Mast cells, eosinophils, and Th2 cell populations were assayed using a flow cytometer. Results: RG products potently inhibited IL-4 expression in phorbol 12-myristate-13-acetate/A23187-stimulated RBL-2H3 cells. Of tested RG products, fRG most potently inhibited IL-4 expression. RG products also alleviated ovalbumin-induced AR in mice. Of these, fRG most potently reduced nasal allergy symptoms and blood IgE levels. fRG treatment also reduced IL-4 and IL-5 levels in bronchoalveolar lavage fluid, nasal mucosa, and reduced mast cells, eosinophils, and Th2 cell populations. Furthermore, treatment with fRG reduced IL-4, IL-5, and IL-13 levels in the colon and restored ovalbumin-suppressed Bacteroidetes and Actinobacteria populations and ovalbumin-induced Firmicutes population in gut microbiota. Treatment with ginsenoside Rd significantly alleviated ovalbumin-induced AR in mice. Conclusion: fRG and ginsenoside Rd may alleviate AR by suppressing IgE, IL-4, IL-5, and IL-13 expression and restoring the composition of gut microbiota.