• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.6
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• pp.1475-1478
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• 2003

Purpose : We hope to evaluate the effectiveness of Citri Reticulatae Viride Pericarpium for the bronchial asthma using assesment on the vascular endothelial growth factor after Citri Reticulatae Viride Pericarpium was intravenously administered OVA-sensitized and -challenged mice. Material and methods: Eleven female mice, 8-10 weeks of age and free of murine specific pathogens, were used. Of the eleven mice, one mouse was not sensitized and ten mice were sensitized by intraperitoneal injection of OVA. Of the sensitized mice, three mice didn't administrate Citri Reticulatae Viride Pericarpium and seven mice administrated Citri Reticulatae Viride Pericarpium. Mice were sensitized on days 1 and 14 by intraperitoneal injection of 20 ㎍ OVA. On days 21, 22, and 23 after the initial sensitization, the mice were challenged for 30 minutes with an aerosol of 1 % OVA in saline. Citri Reticulatae Viride Pericarpium administered 200mg/kg in the tail of the mouse, one time per day, for 7 days, beginning 14 days after first sensitization. Bronchoalveolar lavage was performed 72 hours after the last challenge, and level of VEGF in the BAL fluid were measured by Western blot analysis. Results: Western blot analysis revealed that VEGF protein levels were increased in the all three mice which were challenge with OVA without administered Chung-pi compared the normal mouse. However, in the groups of the administered Chung-pi, the VEGF protein level markedly decreased in six of seven mice. Conclusion : Citri Reticulatae Viride Pericarpium might be effect the treatment of the bronchial asthma as a inhibition of the VEGF.

• Journal of Pharmaceutical Investigation
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• v.31 no.3
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• pp.191-196
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• 2001

Biodegradable polymeric microspheres were studied for their usefulness as carriers for the delivery of vaccine antigens. However, protein antigen could be denatured during microencapsulation processes due to the exposure to the organic phase and stress condition of cavitation and shear force. Therefore this study was carried out to re-evaluate the degree of protein denaturation during microencapsulation with poly(lactide-co-glycolide) (PLGA) copolymer. PLGA microspheres containing ovalbumin (OVA), prepared by W/O/W multiple emulsification method, were suspended in pH 7.4 PBS and incubated with shaking at $37.5^{\circ}C$. Drug released medium was collected periodically and analyzed for protein contents by micro-BCA protein assay. In order to evaluate the protein integrity, release medium was subjected to the analyses of SDS-PAGE and size exclusion chromatography (SEC). And enzyme-linked immunosorbent assay (ELISA) was introduced to measure the immunoreactivity of entrapped OVA and to get an insight into the three-dimensional structure of epitope. The structures of entrapped protein were not affected significantly by the results of SDS-PAGE and SEC. However, immunoreactivity of released antigen was varied, revealing the possibility of protein denaturation in some microspheres when it was evaluate by ELISA method. Therefore, in order to express the degree of protein denaturation, antigenicity ratio (AR) was obtained as follows: amount of immunoreactivity of OVA/total amount of OVA released ${\times}100(%)$. ELISA method was an efficient tool to detect a protein denaturation during microencapsulation and the comparison of AR values resulted in more accurate evaluation for immunoreactivity of entrapped protein.

• BMB Reports
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• v.45 no.5
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• pp.311-316
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• 2012

Sulforaphane (1-isothiocyanato-4-(methylsulfinyl)-butane), belonging to a family of natural compounds that are abundant in broccoli, has received significant therapeutic interest in recent years. However, the molecular basis of its effects remains to be elucidated. In this study, we attempt to determine whether sulforaphane regulates the inflammatory response in an ovalbumin (OVA)-induced murine asthma model. Mice were sensitized with OVA, treated with sulforaphane, and then challenged with OVA. Sulforaphane administration significantly alleviated the OVA-induced airway hyperresponsiveness to inhaled methacholine. Additionally, sulforaphane suppressed the increase in the levels of SOCS-3 and GATA-3 and IL-4 expression in the OVA-challenged mice. Collectively, our results demonstrate that sulforaphane regulates Th2 immune responses. This sutdy provides novel insights into the regulatory role of sulforaphane in allergen-induced Th2 inflammation and airway responses, which indicates its therapeutic potential for asthma and other allergic diseases.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.2
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• pp.343-350
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• 2009

This study aimed to evaluate the anti-asthmatic effects of Samjajihwang-tang (SJT) using OVA-induced asthmatic mice model. Asthmatic mice model was conducted by repeated challenge of OVA using C57BL/6 mice. Each group was treated with distilled water, SJT (400 mg/kg and 200 mg/kg) extract or cyclosporin A (10 mg/kg) for the later 8 weeks, Penh (plethysmography and enhanced pause), immune cells subpopulation, eotaxin, IL-5, TNF-${\alpha}$, Anti-OVA-lgE in BALF (bronchoalveolar lavage), and lung tissue was analyzed, No cytotoxicity of SJT was shown on hFCs (human fibroblast cells). Administration of SJT significantly decreased Penh levels comparing to control group. SJT treatment significantly ameliorated the increase of total cells number and eosinophil including of immune cell subpopulation of $CD3^+/CD69^+$, $CCR3^+$, $B220^+/CD22^+$, $B220^+/CD45^+$, and $B220^+/lgE^+$ cells in BALF comparing to control group. Eotaxin, IL-5, TNF-${\alpha}$, and Anti-OVA-lgE level in BALF were significantly decreased by SJT treatment too. Histopathological finding verified the improvement of infiltration of inflammatory cells and collagen tissue in the SJT groups comparing to control group. These results strongly suggest that SJT would be a effective candidate for herbal-originated anti-asthmatic drug. However, this drug should be further studied for characterization of the accurate action and underlying mechanisms using variant disease model in the future.

• Biomolecules & Therapeutics
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• v.19 no.1
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• pp.77-83
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• 2011

Sulforaphane is an isothiocyanate found in cruciferous vegetables that has been reported to be an effective cancer preventive agent inducing growth arrest and/or cell death in cancer cells of various organs. This paper reports that sulforaphane exerts immunomodulatory activity on the MHC-restricted antigen presenting function. Sulforaphane efficiently increased the class II-restricted presentation of an exogenous antigen, ovalbumin (OVA), in both dendritic cells (DCs) and peritoneal macrophages in vitro. The class II-restricted OVA presentation-enhancing activity of sulforaphane was also confirmed using mice that had been injected with sulforaphane followed by soluble OVA. On the other hand, sulforaphane did not affect the class I-restricted presentation of exogenous OVA at concentrations that increase the class II-restricted antigen presentation. At a high concentration ($20\;{\mu}M$), sulforaphane inhibited the class I-restricted presentation of exogenous OVA. Sulforaphane did not affect the phagocytic activity of the DCs, and the cell surface expression of total H-$2K^b$, B7-1, B7-2 and CD54 molecules, even though it increased the expression of I-$A^b$ molecules to a barely discernable level. These results show that sulforaphane increases the class II-restricted antigen presenting function preferentially, and might provide a novel insight into the mechanisms of the anti-cancer effects of sulforaphane.

• Korean Journal of Animal Reproduction
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• v.16 no.3
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• pp.209-215
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• 1992

These studies were carried out to overcome 2-cell block and in vitro development to blastocysts in vitro fertilization of mouse embryos. The unfertilized ova were obtained by superovulation in ICR mice of 4 to 6 weeks old. Tyrode's 280 solution was used as basal media, and the pH range of media examined was designed from 6.5 to 7.5 with 0.2 interval and the range of osmolality from 250 to 370 mOsm with 20 interval, and the period of sperm preincubation examined was 30, 60, 120, and 180 minutes. The ova developed to 2-cell embryos after 26hrs of incubation with preincubated sperm were evaluatated as in vitro fertilized ones. The results obtained were summarized as follows: 1. The optimal ranges of pH and osmolality of culture media and of sperm preincubation time for in vitro development of in vitro fertilized ova to blastocyst were pH 7.1 to 7.3, 250 to 350 mosmol and 60 to 180 min, respectively. 2. With the media of pH 7.1, 310 mOsm and sperm preincubation period of 120min in another experiment of large sample size, the in vitro fertilized ova was found 66.5% and the in vitro development of in vitro fertilized ova to blastocyst was found 35.8%. From the above results it was concluded that the optimal conditions of pH and osmolality of the media for mouse IVF and embryo culture, and the period of sperm preincubation might be 7.1, 310 mOam and 120min, respectively.

• Molecules and Cells
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• v.22 no.1
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• pp.104-112
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• 2006

We previously reported that DA-9601, ethanol herbal extract of Artemisia asiatica, inhibited histamine and leukotriene releases in guinea pig lung mast cells activated with specific antigen/antibody reaction. This study aimed to evaluate the inhibitory effect of DA-9601 on the OVA-induced airway inflammation in allergic asthma mouse model. BALB/c mice were sensitized and challenged with OVA. DA-9601 was administered orally 1 h before every local OVA-challenge. OVA-specific serum IgE was measured by ELISA, recruitment of inflammatory cells in BAL fluids and lung tissues by Diff-Quik and H&E staining, respectively, the expressions of CD40, CD40L and VCAM-1 by immunohistochemistry, goblet cell hyperplasia by PAS staining, activities of MMPs by gelatin zymography, expressions of mRNA and proteins of cytokines by RT-PCR and ELISA, activities of MAP kinases by western blot, and activity of NF-${\kappa}B$ by EMSA. DA-9601 reduced IgE level, recruitment of inflammatory cells into the BAL fluid and lung tissues, expressions of CD40, CD40L and VCAM-1 molecules, goblet cell hyperplasia, MMPs activity, expressions of mRNA and productions of various cytokines, activities of MAP kinases and NK-${\kappa}B$ increased from OVA-challenged mice. These data suggest that DA-9601 may be developed as a clinical therapeutic agent in allergic diseases due to suppressing the airway allergic inflammation via regulation of various cellular molecules expressed by MAP kinases/NF-${\kappa}B$ pathway.

• The Korea Journal of Herbology
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• v.26 no.4
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• pp.187-194
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• 2011

Objective : Astragali Radix (AR) and Cinnamomi Cortex (CC) are used to enhance immune response in Asian traditional medicine. Immuno-potentiation of the combination of AR and CC were evaluated on the cellular and humoral immune response using murine macrophage cell line (RAW 264.7) and OVA-immunized mice. Methods : This study was designed to investigate the immuno-potentiative effects of AR, CC, and AR with CC on nitric oxide synthesis in RAW 264.7 cells and proliferation and production levels of Intereukin-2 (IL-2) in mouse splenocytes. In addition, we evaluated the plasma-specific antibody responses and splenocyte proliferation on ovalbumin (OVA)-immunized mice treated with herbal extracts. Results : Combination treatment with AR and CC increased nitric oxide synthesis in RAW 264.7 cells and IL-2 level in splenocytes (p<0.001). Combination of AR and CC significantly enhanced the Concanavalin A- (Con A ; T cell mitogen) and lipopolysaccharide-(LPS ; B cell mitogen) induced splenocyte proliferation on the OVA-immunized mice. Combination of AR and CC also significantly enhanced plasma levels of OVA-specific IgG (p<0.01), IgG1 (p<0.05) and total IgM (p<0.01) compared with the OVA-immunized control group. Conclusion : These results suggest that combination of AR and CC could be used as therapeutic profile on activation of immune response.

• Bulletin of the Korean Chemical Society
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• v.23 no.10
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• pp.1413-1431
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• 2002

For the development of immunodetection method of 4,4'-dichlorodipheny-2,2,2-trichloroethane (p,p'-DDT), a persistent and broad toxic organochlorine insecticide, various DDT derivatives were synthesized and characterized for the use of immunogens and the coating ligands for the antibody evaluation. The appropriate lengths of linkers were introduced to investigate more efficient DDT derivatives. Among these hapten derivatives, 2,2-Bis(4-chlorophenyl)acetic acid (DDA), 5,5-Bis(4-chlorophenyl)-5-hydroxypentanoic acid (DDHP) and 5,5-Bis(4-chlorophenyl)-5-chloropentanoic acid (DDCP) were conjugated with keyhole limpet hemocyanin (KLH) for the use of immunogen to produce antibodies. 6,6-Bis(4-chlorophenyl)-6-hydroxyhexanoic acid (DDHH) and 3-[6,6-Bis(4-chlorophenyl)-6-hydroxyhexanoylamino]propanoic acid (DDHHAP) in addition to above hapten derivatives were conjugated to ovualbumin (OVA) and bovine serum albumin (BSA) for the use of coating ligands to measure the titration level of antibody and the displacement of free analytes. Three matching pairs of antibodies and coating ligands were selected for the simultaneous detection of p,p'-DDT and its related compounds of DDA and 2,2-bis(4-chlorophenyl)-1,1-dichloroethylene (p,p'-DDE) by investingating the displacement of free analytes in an indirect ELISA. These were PAb #1 and coating ligand DDCP-OVA, PAb #1 and DDHHAP-OVA, and PAb #3 and DDHHAP-OVA. The most useful immunoreaction for DDT analytes were obtained using PAb #3 and coating ligand DDHHAP-OVA showing 3.4 ng/mL of lower limit of detection. These results indicated that titration level and free analytes displacement were greatly influenced by hapten derivatized and carrier proteins conjugated.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.18 no.3
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• pp.18-25
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• 2005

Objectives: It has a growing interest in the prevention and medical treatment of allergic rhinitis. According to many studies, it's known that Sinicheongpye-um has the inhibitory effect on the allergic rhinitis. We have studied effect of the mice on OVA-induced Production of IL-4, IL-5, $Interferone-{\gamma}$ by Murine Splenocytes and effect of OVA-induced Total IgE. Methods: Mixing Ovalbumin(OVA) $10{\mu}g$ into PBS(phoshate buffered saline) and $Al(OH)_3$, gel solution and changing into $1\;m{\ell}$, we made it into OVA solution. That was administered to normal group. After the last administration into abdominal cavity, we caused allergic rhinitis in nasal cavity of mouse of control group and sample group administering 0.1% solution dropwise 3 times a day for 7 days. Keeping separated serum at -20 degree and after refloating spleen cells, cultivating the cells and centrifuging the upper liquid and keeping it at -20 degree, we measured the amount of IL-4. IL-5, $Interferone-{\gamma}$ and OVA-induced Total IgE by ELISA. Results: 1. In Total IgE, Sinichengpae-um treated group was proved significant inhibitory effect.(p<0.05) 2. In IL-4 study, Sinichengpae-um treated group was proved significant inhibitory effect.(p<0.005) 3. In IL-4 study, Sinichengpae-um treated group was proved significant inhibitory effect.(p<0.001) 4. In $Interferone-{\gamma}$ study, Sinichengpae-um treated group showed a increasing tendency. Conclusion: Based on the above result, it is considered that Sinichengpae-um has the inhibitory effect on the allergic rhinitis of mice and suggested thai it could be used in relieving patients of the symptoms which are caused by allergic rhinitis.